ICBBA 2014 - Université de...

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ICBBA 2014

2nd INTERNATIONAL CONFERENCE ON BIOLOGICAL

& BIOMIMETIC ADHESIVES

6 - 9 May 2014 Istanbul, Turkey

SUPPORTED BY

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COMMITTEES

CONFERENCE SCIENTIFIC COMMITTEE Nick Aldred - Newcastle University, UK Marise Almeida - University of Lisbon, PT Mattias Berglin - University of Gothenburg, SE Manoj K. Chaudhury - Lehigh University, USA Aranzazu Del Campo - Max-Planck-Institut, DE Mehmet S. Eroğlu - Marmara University, TR Patrick Flammang - Mons University, BE Stanislav Gorb - University of Kiel, DE Mustafa O. Güler - Bilkent University, TR Kei Kamino - National Institute of Technology and Evaluation, JP Sabri Kayalı - Istanbul Technical University, TR Philippe Leclere - Mons University, BE Markus Linder - Aalto University, FI Anika Mostaert - University College Dublin, IE Daniel Ruiz-Molina - CIN2, ES Romana Santos - University of Lisbon, PT Willi Schwotzer - Psetta GmbH, CH Alina Sionkowska - Nicolaus Copernicus University, PL Andrew Smith - Ithaca College, USA Russell J Stewart - University of Utah, USA Ayse B. Tekinay - Bilkent University, TR Ebru Toksoy Öner - Marmara University, TR Julius G Vancso - University of Twente, NL

CONFERENCE ORGANIZING COMMITTEE Nick Aldred - Newcastle University, UK Aranzazu Del Campo - Max-Planck-Institut, DE Patrick Flammang - Mons University, BE Stanislav Gorb - University of Kiel, DE Mustafa O. Güler - Bilkent University, TR Markus Linder - Aalto University, FI Anika Mostaert - University College Dublin, IE Romana Santos - University of Lisbon, PT Ayse B. Tekinay - Bilkent University, TR Ebru Toksoy Öner - Marmara University, TR LOCAL ORGANIZING COMMITTEE Ebru Toksoy Öner - Marmara University, TR Mehmet S. Eroğlu - Marmara University, TR Hande Kazak Sarılmışer - Marmara University, TR Songul Yaşar Yıldız - Marmara University, TR Merve Erginer - Marmara University, TR Deniz Köşebent - Marmara University, TR

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TABLE OF CONTENTS

WELCOME 4 SCIENTIFIC PROGRAM 5 MANAGEMENT COMMITTEE MEETING AGENDA 11

PLENARY LECTURES 12

ORAL PRESENTATIONS 33 POSTER PRESENTATIONS 81 INDEX 144

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WELCOME

Dear Colleagues,

On behalf of the Organizing Committee, it is our great pleasure to welcome you to the 2nd International Conference

on Biological and Biomimetic Adhesives & COST Action TD0906 Final Meeting, held on May 6–9 2014 in Istanbul,

Turkey.

The conference has brought together recognized scientists from all around the world from more than 20 countries to

discuss trends and latest technologies in area of bioadhesives. We are very grateful to the support provided by COST,

the Office of Naval Research Global (ONRGlobal) and Marmara University.

Laid out on both Europe and Asia, Istanbul is such a vibrant and fascinating city - ancient and modern, religious and

secular, Asian and European, mystical and earthly, full of contrasts and contradictions. We hope that during the

conference you will enjoy rich scientific program, feel the spirit of Istanbul & experience Turkish hospitality.

We encourage every participant to be active and use this occasion to exchange ideas and to build collaborations and

we hope that your stay in Istanbul will be academically, educationally and socially rewarding.

Cordially yours,

Ebru TOKSOY ÖNER

Marmara University

Department of Bioengineering

Istanbul - Turkey

Patrick FLAMMANG

Université de Mons

Biology of Marine Organisms

and Biomimetics Mons, Belgium

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SCIENTIFIC PROGRAM

Tuesday, May 6, 2014

10:00 - 17:00 Conference Registration

10:00 - 11:00 Opening Ceremony

Opening Speeches

Zafer Gül - Rector of Marmara University

Patrick Flammang - Mons University

Ebru Toksoy Öner - Marmara University

11:00 - 18:00 Panel 1 - Bioinspired Adhesive Designs, Formulations and Applications Chairmen: Markus Linder & Mustafa Guler

11:00 - 11:40 Plenary Lecture 1 - Ali Dhinojwala University of Akron (USA)

Bioinspired Adhesives and Coatings: Lessons Learnt from Spiders and Geckos

11:40 - 12:00

Learning from Protein Design on Biological Adhesives Kei Kamino

National Institute of Technology and Evaluation & Tokyo University (JP)

12:00 - 12:20

Bioadhesion meets functional genomics in flatworms Peter Ladurner

University of Innsbruck (AT)

12:20 - 14:00 Lunch Break

14:00 - 14:20

Water-borne, water-immiscible adhesives inspired by sandcastle worms: sealing fetal membranes after in utero surgery Russell J. Stewart, Sarbjit Kaur, Lovepreet Mann, Ramesha Papanna, Kenneth J. Moise University of Utah (USA)

14:20 - 14:40

Friction under wet conditions: how tree and torrent frogs avoid slipping in their arboreal or watery environments Jon Barnes, Dirk Drotlef University of Glasgow (UK)

14:40 - 15:00

Inspired by algae: studies of alginate/phenol biomimetic adhesives Havazelet Bianco Peled, Ronit Bitton, Yoav Rozen

Israel Institute of Technology (IL)

15:00 - 15:20 Coffee Break

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15:20 - 16:00 Plenary Lecture 2 - Marleen Kamperman Wageningen University (NL)

Physical and Chemical Approaches to Bioinspired Adhesive Design

16:00 - 16:20 Mechanically switchable elastomeric microfibrillar adhesive surfaces Veikko Sariola, Metin Sitti

Carnegie Mellon University (USA)

16:20 - 16:40 A hybrid polymer based on mussel adhesive proteins for dental implantology - BioClou Klaus Rischka, Shahram Ghanaati, Michael Mularczyk, Maria Kozielec, Belma Saldamli, Robert Sader Fraunhofer Institute for Manufacturing Technology and Advanced Materials (Ifam) (DE)

16:40 - 17:00 Combinatorial analysis of nanostructured multilayers films using mussel adhesive inspired polymer Ana I. Neto, Natália L. Vasconcelos, Sara M. Oliveira, João F. Mano University of Minho (PT)

17:00 - 17:20

Bioinspired catechol-terminated self-assembled monolayers with enhanced adhesion properties Mireia Guardingo, Elena Bellido, Jordi Faraudo, Josep Sedó, Albert Verdaguer, Felix Busqué, Daniel Ruiz Molina ICN2-Institut Catala de Nanociencia i Nanotecnologia & CSIC-Consejo Superior de Investigaciones Cientificas (SP)

17:20 - 17:40 Use of biomimetic hexagonal surface texture in friction against lubricated skin Alexey Tsipenyuk, Michael Varenberg Technion (IL)

17:40 - 18:00

Levan-based Adhesive Surfaces for Biomedical Applications

Merve Erginer, Esra C. Mutlu, Mehmet S. Eroğlu, Ebru Toksoy Öner

Marmara University (TR)

18:00 - 19:30 Welcome Cocktail

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Wednesday, May 7, 2014

09:00 - 13:00 Panel 2 - Chemical and Structural Characterization of Natural Adhesives Chairmen: Romana Santos & Kei Kamino

09:00 - 09:40 Plenary lecture 3- Elise Hennebert Mons University (BE)

Deciphering adhesion mechanisms in sea stars

09:40 - 10:00

Integration of surface plasmon resonance imaging in a TIRF microscope, for real-time, label-free imaging of bioadhesion processes Thomas Ederth, Roni Nugraha, Samira Barhemat, Wetra Yandi

Linköping University (SE)

10:00 - 10:20

Fast-track discovery of bioadhesives using a combined high throughput transcritptomics and proteomics approach Ali Miserez, Paul Guerette, Shawn Hoon, Yiqi Seow, Clarinda Sutanto, Fong T Wong Nanyang Technological University (SG)

10:20 - 10:40 Identification of novel marine phenoloxydases by high throughput RNA sequencing Barbara Maldonado, Elise Hennebert, Patrick Flammang, Cecile Van De Weerdt University of Liège & Mons University (BE)


11:00 - 11:40 Plenary Lecture 4 - Marco d'Ischia University of Naples Federico II (IT) Eumelanins and Polydopamine: From Structure-Property Relationships to a Unified Tailoring Strategy

11:40 - 12:00

Comparative study of the attachment of Fucales seaweeds from temperate regions of the Southern Hemisphere Simone Dimartino, David Savory, A. James Mcquillan

University of Canterbury (NZ)

12:00 - 12:20

Investigation of the Adhesion of Levan via Molecular Dynamics Simulations

Binnaz Coşkunkan, Gülçin Cem, Deniz Turgut, K. Yalçın Arga, Deniz Rende, Seyda Bucak, Nihat Baysal, Rahmi Özışık, Ebru Toksoy Öner

Rensselaer Polytechnic Institute (USA), Yeditepe University (TR), Marmara University (TR)

12:20 - 12:40

The projectile slime of velvet worms – Structural characterization of the biological adhesive in Onychophora Alexander Baer, Matthew J. Harrington, Stephan Schmidt, Georg Mayer University of Leipzig (DE)

12:40 - 13:00 Investigations on the protein fraction of Cuvierian tubule adhesive material Demeuldre Mélanie, Wattiez Ruddy, P. Ladurner, Hennebert Elise, Flammang Patrick University of Mons (BE)


13:00 - 14:20 POSTER PRESENTATIONS

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14:20 - 18:20 Panel 3 - Molecular Mechanisms of Bio-Adhesion Chairmen: Patrick Flammang & Stanislav Gorb

14:20 - 15:00 Plenary lecture 5 - Herbert Waite University of California, Santa Barbara (USA)

The Role of Chemistry in the Wet Bioadhesion of Mussels

15:00 - 15:20

Cloning, expression and purification of Nectin, a sea urchin adhesive protein

Duarte Toubarro, Analuce Gouveia, Catarina Alcarva, Raquel Mesquita Ribeiro, Nelson Simões, Gonçalo Conde Da Costa, Carlos Cordeiro, Romana Santos

University of Lisbon (PT)

15:20 - 15:40

Homology but low similarity in adhesive proteins of stalked and acorn barnacles

Anne Marie Power, Jaimie Leigh Jonker, Florence Abram, Ana Varela Coelho, Ingo Grunwald

National University of Ireland (IE)

15:40 - 16:00

Tough Coating Proteins: Subtle Sequence Variation Modulates Cohesion Saurabh Das, Dusty R Miller, Yair Kaufman, Maryte Gylys, Jacob N Israelachvili, Herbert J Waite

University of California, Santa Barbara (USA)


16:20 - 16:40

Cation-pi interaction: New insight for underwater adhesion

Dong Soo Hwang

Pohang University of Science and Technology (KR)

16:40 - 17:00 A new numerical code for hierarchical peeling simulations Lucas Brely, Federico Bosia, Nicola M Pugno University of Torino (IT)

17:00 - 17:20 When the going gets rough... the effect of surface roughness on the attachment abilities of tree frogs

Niall Crawford, Thomas Endlein, Mathis Riehle, W. Jon. P. Barnes University of Glasgow (UK)

17:20 - 17:40 Wet Bioadhesion In Tree Frogs Farzaneh Kaveh, Ciro Semprebon, Michael Kappl, Hans Jürgen Butt Max Planck Institue for Polymer Research (DE)

17:40 - 18:00 One Solution for All Cases: Adhesive Ability of the Leaf Beetle’s Feet Naoe Hosoda, Stanislav N Gorb National Institute for Materials Science (JP), University of Kiel (DE)

18:00 - 18:20 Sticky underwater caddisfly silk: elastic, metal-ion dependent, double network fibers Russell J. Stewart, Nicholas N. Ashton University of Utah (USA)

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Thursday, May 8, 2014

09:00 - 12:00 Panel 4 - Bioadhesion: Prevention Chairmen: Aranzazu Del Campo & Nick Aldred

09:00 - 09:40 Plenary lecture 6 - Nick Aldred Newcastle University (UK)

Biofouling, Bioadhesion and Barnacles

09:40 - 10:20 Plenary lecture 7 – Elisa Martinelli University of Piza (IT)

Surface-nanostructured Amphiphilic Polymers to Prevent the Adhesion and/or Promote the Release of Fouling Organisms

10:20 - 10:40

Where adhesion fails: slippery surfaces of insect-trapping pitcher plants

Ulrike Bauer, Holger Florian Bohn, Walter Federle

University of Bristol (UK)


11:00 - 11:40 Plenary lecture 8 – Paul Molino University of Wollongong (AU)

Microbial Biofouling Slime Layers: Biology, Characterization and the Development of Slime Resistant Coatings

11:40 - 12:00

Study the interaction of cyprid temporary adhesive proteins and surfaces using AFM

Shifeng Guo, Dominik Janczewski, Reddy Sreenivasa Puniredd, Xiaoying Zhu, Serina Siew Chen Lee, Serena Lay Ming Teo, Julius Gyula Vancso

Institute of Materials Research and Engineering A*STAR (SG)

12:00 - 14:00 POSTER PRESENTATIONS


14:00 - 15:40 Panel 5 - Catechol-based Adhesives Chairmen: Russell Stewart & Daniel Ruiz-Molina

14:00 - 14:40 Plenary lecture 9- Phillip B. Messersmith Northwestern University (USA)

Polyphenolic Adhesives and Coatings: Inspiration from Mussels, Tea, Wine, and Chocolate

14:40 - 15:00 Mussel-Inspired Adhesive Interfaces for Biomedical Applications Hakan Ceylan, Samet Kocabey, Hilal Unal Gulsuner, Ayse Begum Tekinay, Mustafa Ozgur Guler Bilkent University (TR)

15:00 - 15:20 Crosslinking kinetics of catecholamine derivatives Julieta Paez, Cristina Serrano, Aránzazu Del Campo Max-Planck-Institut für Polymerforschung (DE)

15:20 - 15:40 Functional hybrid surfaces and interfaces based on bioinspired catechols Daniel Ruiz Molina, Javier Saiz Poseu, Beatriz Garcia, Josep Sedo, Jordi Hernando, Felix Busque Institut Catala de Nanociènica i Nanotecnologia (SP)

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16:00 - 18:00 Panel 6 - Industrials Challenges in Adhesion Chairmen: Alina Sionkowska & Ebru Toksoy Oner

16:00 - 16:40 Plenary lecture 10 - Willi Schwotzer Psetta GmbH (CH)

In Search of Industrial Applications for Biological and Biomimetic Adhesives

16:40 - 17:00

Nature of the problems related to E. coli adhesion on uniform films prepared at low temperature hindering bacterial inactivation: Critical issues S. Rtimi, C. Pulgarin, J. Kiwi Ecole Polytechnique Fédérale de Lausanne(EPFL) (CH)

17:00 - 17:20 Addition Of Tung Oil Diol For Facilitating The Degradation Of Polyurethane Adhesives Jose Miguel Martin Martinez, Pilar Carbonell Blasco University of Alicante (SP)

17:20 - 17:40 Cross-linkable levan derivatives for biodegradable adhesives Ralf Wyrwa, Albrecht Berg, Ebru Toksoy Öner, Matthias Schnabelrauch INNOVENT e. V. (DE) , Marmara University (TR)

17:40 - 18:00 Preparation and properties of alginate mucoadhesive films for oral cavity drug delivery Meir Haber, Irina Lir Biota Ltd. (IL)

20:00 Dinner on Bosphorus

Friday, May 9, 2014

09:00 - 11:00 Action Evaluation


11:20 - 13:00 Action Evaluation

13:00 - 13:30 Closing Ceremony

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MANAGEMENT COMMITTEE MEETING

AGENDA

COST Action no. TD0906 Istanbul, Turkey: May 9th 09:00-13:00

1. Welcome to participants

2. Adoption of agenda

3. Final Evaluation Presentations

a. Action Chair – General Action Presentation

b. Grant Holder – Overview of the Finances of the Action

c. Working Group Presentations by WG Leaders:

i. Work Group 1: Chemical characterization and synthesis of adhesives - Markus Linder (FI)

ii. Work Group 2: Structural characterization of natural and synthetic adhesives Nicholas Aldred (UK)

iii. Work Group 3: Mechanical testing and theory - Stanislas Gorb (DE)

iv. Work Group 4: Fabrication of biomimetic adhesives and their evaluation - Aranzazu Del Campo (DE)

d. STSMs/Dissemination Presentation – Romana Santos (PT)

e. Web coordinator Presentation – Elise Hennebert (BE)

4. Comments from Final Assessment Panel

a. DC Rapporteurs

b. External Experts

5. General discussion

6. AOB

7. Closing

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PLENARY

LECTURES

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Ali Dhinojwala

H. A. Morton Professor in Polymer Science The University of Akron, Akron, OH

Professor Dhinojwala received his Bachelors of Technology Degree in Chemical Engineering from the Indian Institute of Technology,

India and his Ph.D. from Northwestern University in Chemical Engineering in 1994. Thereafter, he was a Research Scientist at the

Department of Materials Science at the University of Illinois, Urbana-Champaign, from 1994 to 1996. At GE Plastics, he worked on

developing plastics for DVDs from 1996 to 1997. In 1997 he joined The University of Akron in the Department of Polymer Science.

Professor Dhinojwala served as a Chair of the Department of Polymer Science from 2008-2012, and is currently H. A. Morton Chair

Professor of Polymer Science. He has been a recipient of the NSF-CAREER Award and NSF-Creativity Award from the National

Science Foundation. He is also a recipient of the Young Faculty Award from 3M Corporation. Professor Dhinojwala is a Fellow of the

American Physical Society.

Professor Dhinojwala’s current research interest is in understanding adhesion, friction, and wetting. His group has developed light-

based spectroscopic techniques to understand the physical properties of molecules at surfaces and interfaces. His recent interest in

the area of bio-adhesion has led them to develop synthetic adhesives inspired by geckos and spiders. He currently has 120 peer-

reviewed publications and ten patents issued or pending.

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BIOINSPIRED ADHESIVES AND COATINGS: LESSONS LEARNT FROM SPIDERS AND

GECKOS

Ali Dhinojwala

The University of Akron, Department of Polymer Science Akron, OH 44325-3909

Insects, spiders, and geckos use brushes of micron or nanometer-size hairs for locomotion or catching preys. Synthetic hairs are

also finding many applications in the areas of dry adhesives, self-cleaning surfaces, field emission displays, and high surface area

coatings for solar cells. I will discuss the properties of the glues used by geckos and spiders and the inspiration it has provided in

designing synthetic adhesives. The role of water and surface chemistry on gecko adhesion will be discussed. This understanding will

help us design reversible adhesives that may work in water and aqueous environments. We have used micro-patterned vertically

aligned carbon nanotubes (CNT) to mimic the micron-size hairs found on the gecko feet. These CNT structures can be transferred to

flexible plastic films to create flexible adhesive tapes. The hierarchical patterns of individual carbon nanotubes (8-10 nm in

diameter) arranged in micron-size patches (50- -cleaning properties of

these synthetic gecko tapes. The application of this technology in thermal management will be discussed. I will also present how

spiders use different architectures of threads for adhesion and a new synthetic design that we have developed to mimic the staple-

pin architecture used by spiders to attach their webs to surfaces.

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Marleen Kamperman

Wageningen University, Physical Chemistry and Colloid Science, Dreijenplein 6, 6703 HB, Wageningen, The Netherlands

Marleen Kamperman is an Assistant Professor at the Physical Chemistry and Colloid Science Laboratory at Wageningen University,

the Netherlands. She received her Ph.D. in Materials Science & Engineering from Cornell University, Ithaca, NY, where she worked

in the group of Prof. Ulrich Wiesner on the development of ordered mesoporous high temperature ceramics using block

copolymers. From 2008 to 2010, she was a postdoctoral researcher in the Functional Surfaces group of Prof. Eduard Arzt at INM –

Leibniz Institute for New Materials in Saarbrücken, Germany, where she worked on the development of bioinspired responsive

adhesive systems. Her research focuses on biologically inspired synthesis of polymers and nanostructured surfaces with controlled

adhesive properties.

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PHYSICAL AND CHEMICAL APPROACHES TO BIOINSPIRED ADHESIVE DESIGN

Marleen Kamperman

Wageningen University, Physical Chemistry and Colloid Science, Dreijenplein 6, 6703 HB, Wageningen

Nature has developed elegant and economical strategies to produce multi-functional and adaptive materials. Drawing inspiration

from a diverse set of natural examples our main aim is to develop new polymeric materials with altered morphologies and

mechanical properties. In this talk I will discuss two projects:

1) Reversible nanostructured adhesive surfaces

Biological attachment systems, found in e.g. geckos, flies and grasshoppers, differ dramatically from conventional adhesives. Unlike

many conventional adhesives that can only be used once on clean, smooth surfaces, and attach accidentally to inappropriate

surfaces, natural so-called "dry adhesives" are reversible, durable, controllable and self-cleaning. The key strategy in dry adhesives

is the incorporation of patterns, i.e. fibrillar surfaces or complex subsurface structures. We developed a nanofabrication processes

based on colloidal self-assembly, which allows the fabrication of elastomers patterned with nanodimples. Compared to smooth

interfaces, adhesion of these nanopatterned surfaces was enhanced, and the pull-off force scaled inversely with dimple amplitude.

All nanopatterned surfaces showed a significant decrease in friction.

2) Catechol containing polymers

3, 4-Dihydroxyphenylalanine (DOPA) is known to play an important role in strong and enduring attachment of biological systems

under wet conditions, such as mussels and sandcastle worms. Much effort has been made to mimic the adhesion properties by

synthesizing polymers containing catechol groups. A common synthesis method is the radical polymerization of vinyl monomers

bearing un-protected catechol groups. Catechol groups are known to be radical scavengers. This scavenging ability may trigger the

catechol to react with the propagating radicals and consequently result in the formation of branched polymer chains. However, in

previous work, the structure of the obtained polymer is always presented as linear polymer chain and little discussion was

presented on the possible side reactions. In this work, we synthesized copolymers of dopamine methacrylamide(DMA) and 2-

methoxyethyl acrylate (MEA) using free radical polymerization with different comonomer ratios. The polymers are characterized

using different techniques, including SEC-MALLS, 1H NMR, DSC and DMA. It was found that the degree of branching indeed

increases with increasing DMA content in the polymer, which explains the poor solubility of homopolymer poly(dopamine

methacrylamide). The adhesion performance of these polymers was evaluated under dry and wet conditions.

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Elise Hennebert

Laboratoire de Biologie des Organisms Marines et Biomimétisme, Université de Mons, Mons, Belgium

Since my Master’s thesis in biological sciences, presented in 2005, my research interests focus on the adhesion mechanisms

developed by sea stars. These marine invertebrates are able to adhere firmly yet temporarily to various substrates using numerous

small locomotive organs called tube feet. The epidermis of the distal part of these organs is specialized for adhesion and encloses

adhesive cells which release an adhesive material upon contact with a surface. My PhD was dedicated to the ultrastructural and

biochemical characterization of this material. I am currently working as F.R.S-FNRS (Belgian Fund for Scientific Research)

Postdoctoral Researcher and my project aims at the identification and characterization of the proteins involved in sea star

adhesion. These proteins are analyzed using biochemical (western blots, mass spectrometry) and molecular techniques (RT-PCR,

transcriptome analysis). The results obtained will allow the production of biomimetic molecules for the development of water-

resistant biomedical adhesives or biomaterials.

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DECIPHERING ADHESION MECHANISMS IN SEA STARS

Elise Hennebert

Laboratoire de Biologie des Organisms Marines et Biomimétisme, Université de Mons, Mons, Belgium

Like every animals belonging to the phylum of Echinodermata, sea stars are characterized by the presence of a water-vascular

system whose visible parts are the tube feet. According to the sea star species considered, these organs may be involved in one or

several of the following functions: locomotion, fixation to the substratum, feeding and burrowing. These different functions are

allowed by the mobility of the proximal part of the tube foot (the so-called stem) as well as by the attachment of the distal part of

the tube foot to the substratum. Tube foot attachment is temporary. Indeed, although tube feet can adhere very strongly to the

substratum, they are also able to detach easily and voluntarily before reinitiating another attachment–detachment cycle. Suction

has long been regarded as a major mean of tube foot attachment in sea stars. However, a number of more recent observations

argue for an adhesive process principally, if not exclusively, mediated by the secretion of an adhesive material.

The comprehension of the complex adhesion mechanisms developed by sea stars requires the complete description of their

secretory organs, the tube feet, the elucidation of the mode of formation of the adhesive material, and the characterization of the

different molecules constituting it. Tube foot morphology and ultrastructure have been investigated in a large diversity of species

while the secreted material has been described and characterized mainly in the species Asterias rubens.

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Marco d’Ischia

Department of Chemical Sciences, University of Naples Federico II,

Via Cintia 4, I-80126 Naples, Italy

Marco d'Ischia is Professor of Organic Chemistry at the Department of Chemical Sciences of Naples University Federico II where he

leads the group of Bioactive and Bioinspired Product Chemistry. He is the author or co-author of over 240 publications in the fields

of melanins and related bioinspired functional materials, including polydopamine, and their application in organic electronics; the

design and mechanism of action of phenolic antioxidants and antinitrosating agents; the oxidative chemistry of catecholamine

neurotransmitters; the oxidation, nitrosation and nitration of biomolecules in relation to oxidative stress-related pathologies. He

has been member of the Council of the European Society for Pigment Cell Research (ESPCR) and Regional Editor of the international

journal Pigment Cell Research (1995-1999). Currently he is: 1) member of the Executive Committee of the Italian Chemical Society,

Division of Organic Chemistry (term of office 2011-2016); 2) Member of the Scientific Committee of the Ischia Advanced School of

Organic Chemistry (IASOC Conference); 3) Scientific coordinator of the European Network for Melanin Research (EuMelaNet). For

his contributions to the chemistry of human pigmentation in 2011 he was awarded the Raper Medal by the International

Federation of Pigment Cell Societies and the European Society for Pigment Cell Research.

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EUMELANINS AND POLYDOPAMINE:

FROM STRUCTURE-PROPERTY RELATIONSHIPS TO A UNIFİED TAILORING STRATEGY

Marco d’Ischia

Department of Chemical Sciences, University of Naples Federico II,

Via Cintia 4, I-80126 Naples, Italy

Polydopamine (PDA), a black insoluble biopolymer produced by autoxidation of the catecholamine neurotransmitter dopamine

(DA), and synthetic eumelanin polymers modeled to the black functional pigments of human skin, hair and eyes have gained

increasing interest in materials science as versatile bioinspired functional systems for a broad range of applications. PDA is

characterized by extraordinary adhesion properties providing efficient and universal surface coating for biomedical, energy, sensing

and environmental applications. Synthetic eumelanins from dopa or 5,6-dihydroxyindoles are the focus of increasing interest as UV-

absorbing agents, antioxidants, free radical scavengers and water-dependent hybrid electronic-ionic semiconductors. Despite

considerable analogies, however, very few attempts have been made so far to provide an integrated unifying perspective of these

two fields of bioinspired, technology-oriented chemical research. As a result, current progress in PDA applications is based more on

empirical approaches than on a solid conceptual framework of structure-property relationships. Developing from a vis-à-vis of PDA

and eumelanin chemistries, this presentation aims to provide an overall view of the various levels of chemical disorder in both

systems and to draw simple correlations with physicochemical properties. Finally it will show how it is possible to translate

chemical knowledge about eumelanin complexity into a unified tailoring strategy for manipulation of eumelanin/PDA structure

through critical control points.

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J. Herbert Waite, PhD

Professor of Biochemistry, University of California, USA

PROFESSIONAL TRAINING

Harvard University, Cambridge, MA Biology/Biochemistry, A.B., 1971

Duke University, Durham, NC Biochemistry/Zoology, PhD., 1976

University of Copenhagen, DK Biochemistry, Post-Doc, 1978

University of Connecticut, Farmington, CT Biochemistry, Post-Doc, 1981

APPOINTMENTS SINCE 1986

Vice-Chair, Graduate Program in BioMolecular Science & Engineering, 2005 to present

Professor, Dept. Chemistry & Biochemistry, UC Santa Barbara, 2004-present

Professor, Dept. Molecular, Cell and Developmental Biology, UC Santa Barbara, 1999-present.

Professor, Chemistry Dept, University of Delaware, 1991-1998

Assoc Professor, College of Marine Sciences & Chemistry Dept, University of Delaware, 1986-1991

RECOGNITION

Award for Excellence in Adhesion Science, Adhesion Society & 3M, 2009

Fellow, American Association for Advancement of Science, 2008

Endowed Harrington Chair in Marine Biochemistry, University of Delaware, 1999

SELECTED PUBLICATIONS

Waite, J.H. & Tanzer, M.L. (1981). Polyphenolic substance of Mytilus edulis: Novel adhesive containing L-Dopa and hydroxyproline.

Science 212, 1039-1040.

Coyne, K.J., Qin, X.X., and Waite, J.H. (1997). Extensible collagen in mussel byssus: A natural block copolymer. Science 277, 1830-

1832.

Harrington, M. J., Masic, A., Holten-Andersen, N., Waite, J. H. and Fratzl, P. (2010). Ironclad fibers: a metal-based biological strategy

for hard flexible coatings. Science 328, 216-220.

J. Yu, E. Danner, R. K. Ashley, Israelachvili, and Waite, J. H. (2011). Mussel protein adhesion depends on interprotein thiol-mediated

redox modulation. Nature Chemical Biology 7, 588-590.

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THE ROLE OF CHEMISTRY IN THE WET BIOADHESION OF MUSSELS

Herbert Waite

Marine Sciences Institute, University of California, Santa Barbara, CA 93106 USA

Organisms and their extended structures are held together by molecular glues whose adhesive and cohesive properties are

adapted to their function. Wet bioadhesion can be specific or opportunistically nonspecific. Mussel adhesion is of the latter type

and depends on elaborately tuned redox chemistry.1 Mussel adhesive proteins contain up to 30 mol% of the catecholic amino acid

DOPA (3, 4-dihydroxyphenylalanine), an exquisitely redox sensitive functionality.1 At or near the interface between the adhesive

proteins and the substratum, mussels impose a highly reducing local environment (low pH and high thiol content) in order to

exploit the superior chemisorption of DOPA to a variety of surfaces.2 Farther from the interface, however, mussel proteins are

maintained in either Fe3+

-rich or oxidizing local environments (high pH and catecholoxidase). The first leads to protein gelation

stabilized by multifunctional DOPA- Fe3+

-complexation, whereas the latter, to the covalent cross-links between DOPAquinone and

reactive amino acids.3-5

By adjusting the redox of the local environment, mussels “tune” the optimal level of adhesion (DOPA) or

cohesion (quinone and cross-linking) needed in each part of the holdfast. A deeper understanding of mussel adhesive chemistry

and its regulation is likely to inspire improvements in adhesive technology especially in wet applications.

References

1- Lee, B. P., Messersmith, P. B., Israelachvili, J. N., Waite, J. H. (2011). Mussel inspired wet adhesives and coatings. Annual Review

of Materials Research 41, 99-132.

2- Yu, J., Wei, W., Danner, E., Ashley, RK, Israelachvili, J., and Waite, J. H. (2011). Mussel protein adhesion depends on interprotein

thiol-mediated redox modulation. Nature Chemical Biology 7, 588-590.

3- Harrington, M. J., Masic, A., Holten-Andersen, N., Waite, J. H. and Fratzl, P. (2010). Ironclad fibers: a metal-based biological

strategy for hard flexible coatings. Science 328, 216-220.

4- Zeng, H., Hwang, D. S., Israelachvili, J. N. and Waite, J. H. (2010). Strong reversible Fe3+

-mediated bridging between dopa-

containing protein films in water. Proc. Nat. Acad. Sci. USA 107, 12850-12853

5- Holten-Andersen, N., Lee, B. P., Messersmith, P. B., Lee, K. Y. C., and Waite, J. H. (2011). pH-induced mussel metal-ligand

crosslinks yield self-healing polymer networks with near-covalent elastic moduli. Proc. Nat. Acad. Sci. USA 108, 2651–2655

23

Nick Aldred

School of Marine Science and Technology, Newcastle University, NE1 7RU, UK

Dr Nick Aldred is a researcher in the laboratory of Prof. Tony Clare at the School of Marine Science and Technology, Newcastle

University, UK. In 2006, after completing his PhD on the subject of the adhesion and adhesives of barnacles and mussels, Nick

began contributing to an on-going project funded by the US Office of Naval Research (ONR), investigating the relation between pre-

settlement behaviour and adhesion of barnacle larvae on experimental fouling-resistant coatings. Having worked subsequently

within EC FP6 and FP7 biofouling-focussed projects AMBIO and SEACOAT, Nick is now co-PI on a new ONR grant that encompasses

evaluation of novel coating formulations using settlement and adhesion assays of algae and invertebrate foulers, as well as direct

investigation of the adhesion and adhesives with a particular focus on barnacle larvae. Using a mutidisciplinary approach that

draws upon techniques including 3-dimensional behavioural analysis, bioassays, proteomics, optical and scanning-probe

microscopy, and analytical spectroscopy, Nick aims to provide coating developers in academia and industry with the information

they need to develop a new generation of environmentally-inert and broadly effective fouling-resistant marine coatings.

24

BIOFOULING, BIOADHESION AND BARNACLES

Nick Aldred

School of Marine Science and Technology, Newcastle University, NE1 7RU, UK

Marine biofouling is the unwanted accumulation of living organisms on structures placed in the oceans by humans. Unsightly it

may be, but the primary drivers for biofouling prevention are seated firmly in economics and environmental awareness. Fouling

significantly accelerates the corrosion of immersed surfaces and, more importantly, results in damage and loss of efficiency for

static (e.g. rigs and sensors) and moving (e.g. ships) structures that are not adequately protected by fouling-resistant coatings.

Maritime shipping is by far the most common and efficient means of bulk transportation, however the hydrodynamic drag penalty

(and therefore wasted fuel) associated with an established hull-fouling community can be up to 80%, with similarly deleterious

effects on manoeuvrability, turning efficiency and carbon emissions. The impact of a simple microbial 'slime' layer can be 10-16%.

A recent analysis of fouling impact in the US Navy surface fleet estimated the cost penalty of biofouling, including cleaning and

efficiency losses, to be in the region of $180-260 million per year. When this figure is extended to the entire global fleet, the figure

quickly multiplies into many billions. In addition to the financial cost, ship-hull fouling is recognised by the International Maritime

Organisation (IMO) as the main vector for translocation of invasive species. The potentially catastrophic effects of introduced

organisms on local ecology are one of the major threats to global biodiversity. It is unsurprising, therefore, that there has been

huge investment by industry and academic researchers into identifying solutions to the scourge of marine biofouling. Usually the

focus in on novel ship-hull coatings and, following the ban on the highly effective but environmentally damaging tributyltin self-

polishing copolymers, focus has shifted clearly towards development of non-biocidal materials to which biofouling organisms, such

as barnacles, simply cannot stick. Commercial products now exist that are effective in repelling settlement of fouling organisms

and/or allow them to be easily removed without the use of biocides. However, these often only work effectively under specific

service regimes. The question is, therefore, how does one develop a material that, without the use of biocides and under variable

conditions, resists adhesion by a wide range of organisms that have honed their attachment strategies to diverse surfaces over

millions of years of evolution? The first half of this presentation will outline recent experimental approaches, their putative

mechanisms of action and future outlook. In the second half, barnacles will be introduced as a model fouling organism and

advances made in understanding the adhesion of barnacle settlement-stage larvae over the course of the COST Action will be

presented. A case will be made, throughout, that we find ourselves at a juncture in the development of fouling-resistant materials,

where future progress and the successful commercialisation of effective, non-toxic, broad-spectrum solutions surely relies upon

improved knowledge of the adhesive mechanisms used by the attaching organisms.

25

Elisa Martinelli

Department of Chemistry and Industrial Chemistry and INSTM UdR Pisa

University of Pisa, Italy, via Risorgimento 35, 56126 Pisa, Italy

Dr. Elisa Martinelli received her PhD in Chemical Sciences in 2008 at the University of Pisa, Italy, where she now works in

collaboration with Professor Galli since several years.

Her research activities are focused on the design and synthesis of nanostructured polymers and nanocomposites, which show

potential for innovative environmentally benign applications. In particular, she has been involved in EU-funded projects on

prevention and release of biofouling, in which she has contributed significant papers and industrial patents.

26

SURFACE-NANOSTRUCTURED AMPHIPHILIC POLYMERS TO PREVENT THE

ADHESION AND/OR PROMOTE THE RELEASE OF FOULING ORGANISMS

Elisa Martinelli, Giancarlo Galli

Department of Chemistry and Industrial Chemistry and INSTM UdR Pisa

University of Pisa, Italy, via Risorgimento 35, 56126 Pisa, Italy

Tuning of the properties at a molecular level of the nanostructured surface of a polymer film may lead to breakthroughs in the field

of marine biofouling where interfacial interactions, operating within a few nanometers of a surface, are critical. For example, larvae

of invertebrates and spores of algae are highly selective in their preferences for certain surfaces. Several interfacial properties of

the surface have been shown to act either as cues that moderate initial settlement of the fouling organism, or as factors that

determine adhesion strength.1

Recent trends to combat marine biofouling by means of non-biocidal coatings include the use of hybrid polymer nanocomposites,

biomimetic zwitterionic polymers, polymer-analogue platforms and phase-segregated siloxane polyurethane copolymers.1 All these

‘green’ technologies share the objective to prevent biofouling through manipulation of the physicochemical and/or materials

properties of the coating, so that the organism either perceives the surface as unconducive to settlement or the intermolecular

interaction forces between the surface and the polymeric adhesives produced by the fouling organism are weakened, promoting

adhesive failure.

In our approach2–4

we devised novel low elastic modulus and low surface energy coatings by the homogeneous dispersion of

variously engineered surface-active fluorinated polymers in a polymer matrix. We show that the surface morphological, topological

and compositional nano- to micro-scale complexities of the surface-segregated polymer film add synergistically to enhance release

of marine biofouling organisms in laboratory bioassays and field test trials. In particular, incorporation of amphiphilic polymers into

an elastomeric polymer matrix results in distinct biological performances against micro- and macro-organisms with contrasting

tendencies to interact with the substratum.

References

1- J.A. Callow, M.E. Callow, Nature Commun., 244 (2011).

2- E. Martinelli, M. K. Sarvothaman, G. Galli, M. E. Pettitt, M. E. Callow, J. A. Callow, S. L. Conlan, S. A. Clare, A. B. Sugiharto, C.

Davies, D. Williams, Biofouling, 28, 571 (2012).

3- M. Atlar, B. Ünal, U.O. Ünal, G. Politis, E. Martinelli, G. Galli, C. Davies, D. Williams, Biofouling, 29, 39 (2013).

4- B. R. Yasani, E. Martinelli, G. Galli, A. Glisenti, S. Mieszkin, M. E. Callow, J. A. Callow, Biofouling, (2014)

http://dx.doi.org/10.1080/08927014.2013.878864.

27

Paul Molino

Intelligent Polymer Research Institute, ARC Centre of Excellence for Electromaterials Science, University of Wollongong,

Wollongong, NSW 2522, Australia

Dr Paul Molino completed his PhD in Botany at the University of Melbourne, Australia in 2008. He is currently a Vice Chancellor

Research Fellow at the Intelligent Polymer Research Institute (IPRI), ARC Centre of Excellence for Electromaterials Science, at the

University of Wollongong, Australia. His research interests include studying fundamental biomolecular and cellular interactions

with nanostructured materials using a range of microscopy techniques and microgravimetric surface sensing techniques. He is

interested in the design of biomaterials to promote beneficial protein and cellular interactions, as well as developing ultra-low

fouling materials and coatings for both biomedical and environmental applications.

28

MICROBIAL BIOFOULING SLIME LAYERS: BIOLOGY, CHARACTERISATION AND THE

DEVELOPMENT OF SLIME RESISTANT COATINGS

Paul Molino

Intelligent Polymer Research Institute, ARC Centre of Excellence for Electromaterials Science, University of Wollongong,

Wollongong, NSW 2522, Australia

Microbial slime layers, almost exclusively composed of bacteria and diatoms, plague many modern fouling resistant coating

systems, forming a biological layer on the coating surface that provides an attractive surface for colonisation by secondary fouling

organisms. Biofouling diatoms can engage a number of mechanisms to adhere and persist on both toxic and environmentally

friendly antifouling coatings, enabling them to be the dominant group in these slime layers that alone are a major problem for

several maritime based industries. Here I will discuss the fundamental biology of biofouling diatoms, including the adhesion

strategies that make them such successful biofouling organisms on modern fouling release marine coatings. Additionally I will

present some modern characterisation techniques that are being used to provide insights into the nature of diatom based

adhesives and cellular interactions with surfaces, and how this is guiding the development of slime resistant coatings.

29

Phillip B. Messersmith

Departments of Biomedical Engineering, Materials Science and Engineering and Chemical and Biological Engineering

Chemistry of Life Processes Institute, Institute for Bionanotechnology in Medicine, and Robert H. Lurie Comprehensive Cancer

Center, Northwestern University, Evanston, IL, USA

Phillip B. Messersmith is the Erastus O. Haven Professor at Northwestern University. He is appointed in the Departments of

Biomedical Engineering, Materials Science and Engineering and Chemical and Biological Engineering. He earned his B.S. degree in

life sciences in 1985 from the University of Illinois at Urbana, M.S degree in bioengineering from Clemson University, and his Ph.D.

degree in materials science and engineering in 1993 from the University of Illinois at Urbana. Dr. Messersmith was a postdoctoral

fellow at Cornell University from 1993-1994 and a faculty member at the University of Illinois at Chicago from 1994-1997. His

awards and honors include a MERIT award from the National Institutes of Health, the Langmuir Lecture Award from the American

Chemical Society, and the 2013 Clemson Award for Basic Research from the Society for Biomaterials. Dr. Messersmith is a fellow of

the American Institute for Medical and Biological Engineering, the Royal Society of Chemistry, and the International Union of

Societies of Biomaterials Science and Engineering. Dr. Messersmith is co-Editor in Chief of the Royal Society of Chemistry journal

Biomaterials Science, and is a member of the editorial/advisory boards of Soft Matter, Langmuir, Materials Horizons,

Nanomedicine, Biointerphases, Biomedical Materials, and Bioinspired, Biomimetic and Nanobiomaterials. His research interests

include studies of biological adhesives, including mussel adhesive proteins and gecko adhesives, the design of biomimetic adhesive

polymers and polymer composites, development of novel biomaterials for regenerative medicine, and antifouling polymer surfaces.

30

POLYPHENOLIC ADHESIVES AND COATINGS: INSPIRATION FROM MUSSELS, TEA,

WINE, AND CHOCOLATE

Phillip B. Messersmith

Departments of Biomedical Engineering, Materials Science and Engineering and Chemical and Biological Engineering

Chemistry of Life Processes Institute, Institute for Bionanotechnology in Medicine, and Robert H. Lurie Comprehensive Cancer

Center, Northwestern University, Evanston, IL, USA

Polyphenols are found in both plant and animal tissues, where they serve a variety of functions including mechanical adhesion,

structural support, pigmentation, radiation protection, and chemical defense. In animals, polyphenols are found in the adhesive

proteins secreted by sessile marine organisms. In mussels, the adhesive proteins are known to contain high levels of 3,4-dihydroxy-

L-alanine (DOPA), an amino acid that is believed to be important in adhesion to substrates. In plants, polyphenolic compounds

containing benzenediol (catechol) and/or benzenetriol (gallol) functional groups are widely distributed secondary metabolites with

a variety of biochemical and physical functions. Consumption of foods and beverages rich in polyphenols are claimed to be

beneficial to one’s health.

This talk will focus on selected biological polyphenols that are rich in catechol or gallol functional groups, with the goal of

developing novel materials inspired by biological polyphenols. In the case of mussel-inspired biomaterials, we are interested in

understanding the molecular and mechanochemical aspects of mussel adhesion, and in developing biomimetic polymer hydrogels

and coatings from synthetic catechol containing polymers. These biologically inspired materials have a variety of functional uses,

including tissue repair, drug delivery and antifouling coatings. In the case of plant polyphenols, we recently reported the formation

of thin adherent polymerized films on substrates immersed in aqueous extracts of tea, coffee beans, cacao beans and grapes.

Deposition is facile on a variety of solid, porous and nanoparticulate substrates composed of metals, ceramics and polymers. In

addition to possessing inherent antibacterial and antioxidant properties, the deposited polyphenol films serve as versatile ‘primers’

facilitating secondary modifications of the primer coating such as metallization and covalent grafting of biomolecules and synthetic

polymers. These secondary modifications can be exploited for a variety of practical applications, including antibacterial,

antioxidant and fouling resistant coatings on medical devices, metal deposition, plasmonic tuning and surface functionalization of

nanoparticles,

31

Willi Schwotzer

Psetta GmbH

Willi Schwotzer is a chemist by training. For many years he was CTO (Chief Technology Officer) of Collano. This Swiss group of

companies is active in R&D, production and marketing of industrial adhesives for various applications. After retiring in 2013 he

founded Psetta GmbH, a consulting firm for knowledge communication (www.psetta.ch). He also is co-chairman of S-WIN, the

Swiss Wood Innovation Network (www.s-win.ch).

32

IN SEARCH OF INDUSTRIAL APPLICATIONS FOR BIOLOGICAL AND BIOMIMETIC

ADHESIVES

Willi Schwotzer

Psetta GmbH

Cost Action TD0906 revealed a fascinating wealth of biological adhesives and adhesive bonding mechanisms. Their high

performance level raises hope for their industrial use in technically challenging applications. However, in reality their commercial

contribution to the total adhesive market is very small by any definition. How come?

In order to understand this apparent paradox, a closer look at main differences between biological and man-made adhesives is

required. Specifically, three areas have to be scrutinized:

The principles behind the evolution and development of biological and man-made adhesives, respectively.

The scale in which biological and man-made adhesives are produced and applied.

The requirements for the (thermodynamic) stability of adhesives depending on the intended application.

Examples of commercial applications of biological and biomimetic adhesives principles indicate that such systems can be only

successful if one or more of the following conditions are overriding the cost issue:

Repeated bonding/de-bonding on demand

Bonding to wet surfaces

Adhesion to disparate substrates

Easy-to-use, dry and non-stick systems

Production of the biomimetic adhesive by methods of industrial mass production

Such conditions are most likely met in the following fields of application:

Adhesives for use during surgery

Adhesives for targeted drug delivery

Films, foils and webs for temporary fixations (viz., with bonding/de-bonding properties)

In summary it is safe to assume that biological and biomimetic adhesives are geared for a use as high-end product in niche markets.

Such markets are generally very attractive because of the high added value.

33

ORAL

PRESENTATIONS

34

Panel 1

Bioinspired Adhesive Designs, Formulations and Applications

35

OP-01 LEARNING FROM PROTEIN DESIGN ON BIOLOGICAL ADHESIVES

Kei Kamino

Natl. Inst. Tech. Eval. & Tokyo Univ. Sci.

Attachment of materials in water is challenging. If the technology is practical, innovation in manufacturing processes, medical

technologies, and our daily life will be far beyond our expectation. Mechanisms of attachment in water are actually complex.

Sessile organisms ranging from microbes to hard and soft animals and plants have developed diversified ways to tightly and

continuously attach to several material surfaces. These biological adhesives are excellent models from which to learn how to

artificially attach materials in water and to obtain information that will be useful to develop general theories in the interface

sciences. Thus, we continue to study them, particularly to characterize the adhesive’s native proteins. Hopefully, combination of

chemical, biological and physical characterization of the native adhesives with material design for the practice would be significant.

However, proteins in the biological adhesives and synthetic simple polymers are miles apart actually. This author think that protein

based materials such as modular proteins may arrange a match between them. In this paper, barnacle adhesive proteins / peptides

and peptides in mussel foot proteins and fungal cellulose binding module (CBM) were linked via simple linker sequence. Bacterial

recombinant forms of the proteins were purified in physiological conditions and the proteins were characterized with ToF-MS, GFC,

QCM-D, and AFM. The modules used in this study included Megabalanus rosa cement protein(Mrcp)-20kDa, Mrcp-19kDa, amyloid-

like peptide in Mrcp-52kDa, repetitive unit in mussel fp1 and fp2, and fungal CBM.

36

OP-02 BIOADHESION MEETS FUNCTIONAL GENOMICS IN FLATWORMS

Peter Ladurner

Institute of Zoology, University of Innsbruck, Austria

Background-Aim: Many organisms use adhesives to attach to substrates but the molecular nature of these glues is not well

understood. Our current efforts are aiming to unravel the molecular basis of biological adhesion of the free-living flatworm

Macrostomum lignano (Platyhelminthes). The adhesive organs of M. lignano consist of a duo-gland system comprising three cell

types, an adhesive gland, a releasing gland, and a modified epidermal cell called anchor cell. M. lignano possesses about 130

adhesive organs, which enable the worm to adhere to and release very rapidly from the substrate in seawater.

Results-Methods: We analysed in detail the morphology of the adhesive organs using light- and electron microscopy. Next, we

generated a tail-specific transcriptome using differential gene expression resulting in an adhesion related candidate gene list of

about 250 genes. In the following step we analysed the spatial expression of 239 of these genes by a whole mount in situ

hybridization screen. Using messenger RNA knock-down by RNA interference we analysed the biological function of a subset of

these genes. Polyclonal antibodies were generated against selected proteins.

Conclusion: Our experiments corroborate that a molecular biology approach allows the identification of adhesion related proteins

and can contribute to understand the underlying adhesive mechanism.

Supported by FWF grant P25404-B25 and COST Action TD0906

M_lignano_adhesive_organ_cross section ELMI

Electron microscopical cross section of an adhesive organ of the flatworm Macrostomum lignano. A collar of 22 microvilli with a

dense actin core surrounds the neck of an adhesive gland with a large electron-dense adhesive vesicle, and a neck of a releasing

gland cell containing smaller electron lucent vesicles.

37

OP-03 WATER-BORNE, WATER-IMMISCIBLE ADHESIVES INSPIRED BY SANDCASTLE

WORMS: SEALING FETAL MEMBRANES AFTER IN UTERO SURGERY

Russell J. Stewart1, Sarbjit Kaur

1, Lovepreet Mann

2, Ramesha Papanna

2, Kenneth J. Moise

2

1Department of Bioengineering, University of Utah, Salt Lake City, Utah, USA 2Texas Fetal Center, The University of Texas, Houston, Texas, USA

Background: There remains room for improvement of tissue adhesives. In an approach inspired by marine sandcastle worms,

water-borne adhesives were formed by associative macro phase separation (complex coacervation) of synthetic copolyelectrolytes

(co-PEs) that mimic adhesive proteins of natural sandcastle worm glue. The injectable adhesive complex coacervates can be applied

to and bond fully-submerged substrates.

Methods: Polymethacrylate co-PE bioanalogs were synthesized by free radical co-polymerization. Methacrylate groups were

conjugated to the co-PEs for covalent crosslinking (curing). The adhesives were tested for sealing fetal membranes (FM) defects in

pregnant mini-pigs. Tissue scaffold plugs were inserted into trocar created defects and sealed with chemically cured adhesive. FMs

were retrieved after 3 wks for histology.

Results: Curing through methacryloyl side chains was rapid and efficient by both chemical and photo initiation. The final modulus

and bond strength of cured adhesives could be modulated over a wide range by any of several means: concentration or intensity of

initiator, mol% of methacryoyl side chains, inclusion of solid fillers, and by second polymer networks within the coacervated co-PE

network. The adhesives adhere strongly to several types of common biomaterials. Crosslinked adhesives swelled <1% over 30 days

in physiological saline. With adhesive, the plugs were present in FM defects after 3 wks. Without adhesive the plugs were

dislodged. There were no adverse effects on fetus or mother.

Conclusion: Adhesive complex coacervates show promise as a means to seal human FM defects after fetal surgeries, which may

allow in utero fetal interventions for additional indications in the future.

38

OP-04 FRICTION UNDER WET CONDITIONS: HOW TREE AND TORRENT FROGS AVOID

SLIPPING IN THEIR ARBOREAL OR WATERY ENVIRONMENTS

Jon Barnes1, Dirk Drotlef

2, Walter Federle

3

1Centre for Cell Engineering, University of Glasgow, Scotland, UK 2Max-Planck-Institut fuer Polymerforschung, Mainz, Germany 3Department of Zoology, University of Cambridge, UK

Aided by their specialised toe pads, tree and torrent frogs combine good adhesion and friction with effortless detachment while

climbing in their wet environments. They are an excellent model for reversible and multi-use adhesives that could adhere in wet

conditions. The dominant mechanism of adhesion is capillarity, there being a thin layer of fluid separating the frog’s pad from the

substrate. Adding fluid usually leads to a reduction in friction. How then do these frogs maintain good friction under such

conditions?

Measurements of forces generated by individual toe pads of intact frogs show that, in spite of the fluid layer, toe pads generate

static friction. This is demonstrated by the build-up of force at the onset of sliding and the presence of remaining shear forces two

minutes after sliding stopped. Measured shear forces are too large to be explained by the deformation of the fluid film’s meniscus.

Instead, they are due to the formation of dry contacts between pad and substrate. Measurements of the thickness of the fluid layer

under the pad using interference reflection microscopy support this hypothesis, mean fluid layer thickness in the central regions of

each epithelial cell being <5nm.

Toe pads have a multilayered structure, the outer layer consisting of hexagonal cells separated by narrow channels. The surface of

these cells is covered by a dense array of nanopillars which underlie the observed static friction. Here we review the structure and

physical properties of these nanopillars, analysing their importance in the dynamics of toe pad function.

Toe pad structure in a tree frog

a, White's tree frog; b, adhesive toe pad; c, toepad epithelial cells;, d,e, SEM and TEM of nanopillars that cover the epithelial cells.

39

OP-05 INSPIRED BY ALGAE: STUDIES OF ALGINATE / PHENOL BIOMIMETIC ADHESIVES

Havazelet Bianco Peled, Ronit Bitton, Yoav Rozen

Department of Chemical Engineering, Technion - Israel Institute of Technology, Haifa, Israel

Background-Aim: Adhesives that functions well under moist conditions could facilitate many surgical procedures. A source of

inspiration for their design are algae, known in remarkable adherence to wet surfaces. Our previous studies revealed that the glue

secreted by brown algae is composed of alginate gel and polymeric phenols. The overall aim of the current research is to lay the

basis for the development of a new family of bioinspired sealants based on alginate and monomeric phenolic compounds.

Materials-Methods: adhesive formulations were prepared from mixtures of alginate, divalent ions and phenols. Adhesion was

assessed using lap-shear and tensile assays as well as custom-made apparatuses. Gel point was determined using rheology and

nanostructure was evaluated using SAXS.

Results (quantitative and / or statistical data): Adhesion strength showed a strong dependence on the source of the calcium cross-

linker, the alginate G-content, the molecular weight of the alginate and the viscosity. Phenols altered both the gel point and the

adhesion however did not affect the Young modulus or the nanostructure of the cured adhesive.

Discussion: Cohesion strength is directly related to the properties of the cross-linked alginate while the main contribution of the

phenols is manifested in the sealant–substrate interaction. Evidence of the importance of phenol structure is presented.

Conclusion: The adherence of the biomimetic adhesives is mainly due to mechanical interlocking. Preliminary evidence suggests

that the size and the arrangement of hydroxyl groups on the benzene ring might affect sealant performance.

40

OP-06 MECHANICALLY SWITCHABLE ELASTOMERIC MICROFIBRILLAR ADHESIVE SURFACES

Veikko Sariola, Metin Sitti

Carnegie Mellon University

Background-Aim: Our goal is to develop adhesive surfaces that can be controllably switched off for easy release. Here we propose

a surface that consists of two components: a thin non-adhesive mesh and elastomeric pillars, covered with a gecko-inspired

microfibrillar adhesive, extending through the holes in the mesh. The switching is achieved by retracting the pillars.

Materials-Methods: The pillars and the microfibrillar adhesive were cast using polyurethane. The non-adhesive mesh was laser cut

from an acetal sheet. Motorized stages controlled the relative distance of the two surfaces. The surfaces were characterized by

loading and unloading on glass substrates.

Results: The pull-off pressure of the adhesive was 15 kPa. With the adhesive switched off, no pull-off force could be observed

within the resolution of our load cell. From this, we estimated switching ratios of at least 2000-to-1. We demonstrated that the

surface can be used for picking and placing silicon chips, kapton sheets and various other parts.

Discussion: The switching ratio is likely higher, because we could not measure the tiny forces when the adhesive was turned off.

The surface is not limited to geckoadhesives: any reusable adhesive coating can be used. The surface can only release parts that are

larger than one mesh hole (currently 2.6 mm).

Conclusion: The switching ratios are among the highest ever reported, the switching method is easy and reliable, and the surface

can be used for robotic pick-and-place applications. Future work will focus on miniaturization of the structures and integrated

actuation mechanisms.

Schematics and images of the mechanically switchable structured adhesive surface

Schematics and images of the mechanically switchable structured adhesive surface. a) Illustration of the switching principle: I:

microscopic dry microfibrillar adhesive, II: macroscopic elastomer pillars, III: non-adhesive mesh. The adhesive is ON when the pillars

are extended through the holes in the mesh. b) By retracting the pillars, the adhesive can be turned OFF. c) Photograph of the real

surface. Labeling is same as before. d) SEM micrograph of the dry microfibrillar adhesive. The microfibers have mushroom shaped

tips.

41

Characterization of the switchable adhesive surface

Characterization of the switchable adhesive surface. Pull-off force measured in 12 cycles of adhesion ON and 12 cycles, where the

adhesive is first preloaded against a substrate and then switched OFF. The switching ratio is at least 2000-to-1 and switching is very

reliable and repeatable.

Veikko Sariola was supported by Jenny and Antti Wihuri Foundation, Walter Ahlström foundation, and the Academy of Finland

(grant 268685), and Metin Sitti was supported by the NSF CMMI-1130520 grant.

42

OP-07 A HYBRID POLYMER BASED ON MUSSEL ADHESIVE PROTEINS FOR DENTAL

IMPLANTOLOGY - BIOCLOU

Klaus Rischka1, Shahram Ghanaati

3, Michael Mularczyk

4, Maria Kozielec

1, Belma Saldamli

5, Robert Sader

2

1Fraunhofer Institute for Manufacturing Technology and Advanced Materials (Ifam), Bremen, Germany 2Department of Oral and Maxillofacial Surgery, University of Frankfurt, Germany 3Institute of Pathology, University of Mainz, Germany 4Federal Institute for Materials Research and Testing (Mpa), Technical University of Darmstadt, Germany 5Clinic for Orthopedics and Sports Orthopedics, Technical University of Munich, Germany

Background-Aim: Preserving the gum’s natural barrier function at the point of intervention is of vital importance in preventing

bacterial penetration, which can cause local infection and subsequent bone loss, and is therefore the deciding factor for the success

and long term stability of a dental implant.

In contrast to the natural teeth, the biological width around tooth implants is loosely attached and does not have direct contact to

the biological material.

The goal was the development of a biocompatible bonding agent based on the mussel adhesive protein Mefp-1 for the fixation of

dental implants.

Materials-Methods: The adhesion Mefp-1-peptides were synthesized by solid-phase-peptide-synthesis and combined with a

polymer afterwards.

The biocompatibility and the bonding properties of the adhesive were determined in a key experiment. The in vitro tests were

performed according to DIN EN ISO 10993 with the animal fibroblast cell line L-929 and primary human fibroblasts. In vitro studies

were performed on a subcutane implantation model on CD-1 mice. In order to determine tensile strength, pig gingivas were

bonded to titanium with the target biomaterial.

Additionally the adsorption properties of the peptides were evaluated by XPS and QCM-D.

Conclusion: It can be stated that this biomaterial shows excellent biocompatibility with a very low manifestation or induction of

inflammatory cell components. Furthermore, the peptide modification of the biomaterial improved bonding properties in the

model

43

Cytotoxicity of a DOPA-containing peptide

Vitality of primary human fibroblasts against a DOPA-containing peptide

XPS-Measurement

XPS measurements of a Ti-platelet following immobilisation of the Mefp-1-decapeptides from the MOPS buffer (after 24 hours, c = 1

mg/ml)

44

OP-08 COMBINATORIAL ANALYSIS OF NANOSTRUCTURED MULTILAYERS FILMS USING

MUSSEL ADHESIVE INSPIRED POLYMER

Ana I. Neto, Natália L. Vanconcelos, Sara M. Oliveira, João F. Mano

3B 's Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, AvePark, 4806-90 Taipas, Guimarães, Portugal; ICVS/3B

's PT Government Associate Laboratory, Braga/Guimarães, Portugal

Background-Aim: In a marine environment, specific proteins are secreted by mussels and used as a bioglue to stick to a surface

allowing generate irreversible bonding. Inspired by the structure and properties of mussel adhesive proteins, layer-by-layer (LbL)

coatings based on polymers that contain catechol groups were developed.

Materials-Methods: We used dopamine-modified hyaluronic acid (HA-DN) prepared by carbodiimide chemistry to form thin and

surface adherent dopamine films. The multilayer films were developed by electrostatic interactions using chitosan (CHT) as

polycation and HA-DN, HA-4DN, HA and alginate (ALG) as polyanions. The formation of these films was investigated in-situ by

quartz crystal microbalance with dissipation monitoring (QCM-D). Many combinations of the marine inspired biomaterials were

analyzed in a high-throughput way. Films with different number of layers were constructed and individually disposed on isolated

transparent spots, patterned onto biomimetic superhydrophobic substrates. The coatings with different number of layers were

characterized by atomic force microscopy and scanning electron microscopy.

Results-Discussion: The adhesion properties of the resulting films in the chips were measured showing that the nanostructured

films of the conjugates promote a better adhesion when compared with CHT/HA and CHT/ALG films. Individual spots containing

different number of layers were modified by including drops of protein and crosslinking reagent and in vitro tests using two distinct

cell sources were conducted.

Conclusion: The results showed an enhanced cell adhesion for the biomimetic films that contain catechol groups, demonstrating

their potential to be used in different biomedical applications, including tissue engineering.

45

OP-09 BIOINSPIRED CATECHOL-TERMINATED SELF-ASSEMBLED MONOLAYERS WITH

ENHANCED ADHESION PROPERTIES

Mireia Guardingo1, Elena Bellido

1, Jordi Faraudo

2, Josep Sedó

1, Albert Verdaguer

1, Felix Busqué

3, Daniel Ruiz Molina

1

1ICN2-Institut Catala de Nanociencia i Nanotecnologia. Campus UAB, 08193 Bellaterra (Barcelona), Spain; CSIC-Consejo Superior de Investigaciones

Cientificas. ICN2 Building, Campus UAB, 08193 Bellaterra (Barcelona), Spain 2Institut de Ciència de Materials de Barcelona (CSIC). Campus UAB, 08193 Cerdanyola del Vallès (Barcelona), Spain 3Universitat Autònoma de Barcelona. Campus UAB, 08193 Cerdanyola del Vallès (Barcelona), Spain

Background-Aim: The role of the catechol moiety in the adhesive properties of mussel proteins and related synthetic materials has

been extensively studied over the last years but still remains elusive. Here, a simplified model approach is presented based on a

self-assembled monolayer of upward-facing catechols thiol-bound to epitaxial gold substrates.

Materials-Methods: SAMs of 4-(6’-mercaptohexyl)catechol were obtained on gold substrates. Afterwards, an Atomic Force

Miscroscope was used to locally measure the adhesion by means of Force-distance curves.

Results (quantitative and / or statistical data): The catechol-modified surfaces showed an average adhesion force of 45 nN. The

interaction proved to be slightly stronger, more reproducible and less statistically disperse than a reference polydopamine coating.

Further studies revealed the influence of the surface roughness on the adhesive properties of the SAM. The interaction of the AFM

tip with a catechol-modified polycrystalline gold substrate was not measurable but these substrates were able to assemble

magnetic nanoparticles.

Finally, the influence of the catechol group on the formation and quality of the SAM was explored both theoretically and

experimentally using direct-write AFM lithography.

Discussion: The superior chemical and topographical homogeneity of the SAM with respect to a polydopamine coating is

considered the main reason for its enhanced adhesion. Also, the SAM proved to be more robust than the polydopamine coating

due to its covalent bonding to the surface.

Conclusion: SAMs of catechol are shown to be an excellent and effective alternative to polydopamine yielding more homogenous

substrates with enhanced adhesive properties.

46

Histogram

Histogram obtained from the adhesion measurements on a PDA coating (solid purple bars) and a catechol-terminated SAM (striped

black bars). The monolayer shows diminished dispersion in the results and slightly higher adhesion force due to its homogeneity and

high density of adhesive moieties.

Mercaptohexyl-catechol

47

OP-10 USE OF BIOMIMETIC HEXAGONAL SURFACE TEXTURE IN FRICTION AGAINST

LUBRICATED SKIN

Alexey Tsipenyuk, Michael Varenberg

Dept. of Mechanical Engineering, Technion, Haifa, Israel

Smooth contact pads evolved in insects, amphibians and mammals to enhance the attachment abilities of the animal’s feet are

often dressed with surface micropatterns of different shapes that act in the presence of a fluid secretion. One of the most striking

surface patterns observed in contact pads of these animals is based on a hexagonal texture, which is recognized as a friction-

oriented feature capable of suppressing both stick-slip and hydroplaning whilst enabling friction tuning. Here we compare this

design of natural friction surfaces to textures developed for working in similar conditions in disposable safety razors. When slid

against lubricated human skin, the hexagonal surface texture is capable of generating about twice the friction of its technical

competitors, which is related to it being much more effective at channeling of the lubricant fluid out of the contact zone. The

draining channels shape and contact area fraction are found to be the most important geometrical parameters governing the fluid

drainage rate.

48

OP-11 LEVAN-BASED ADHESIVE SURFACES FOR BIOMEDICAL APPLICATIONS

Merve Erginer1, Esra C. Mutlu

1, Mehmet S. Eroğlu

2, Ebru Toksoy Öner

1

1Marmara University, Department of Bioengineering, Istanbul, Turkey 2Marmara University, Department of Chemical Engineering, Istanbul, Turkey

Levan is a homopolymer of fructose with many unique properties like high solubility in oil and water, good biocompatibility and

film-forming ability. One of the outstanding properties of this polysaccharide is its strong bioadhesivity 1,2

. Hydroxyl groups in its

structure form adhesive bonds with various substrates2. Levan-based bioactive surfaces hold great importance for their potential

uses in Biotechnology sector, especially for Tissue Engineering and Biomedical applications. In fact, a recent literature analysis on

microbial exopolysaccharides attributed levan together with xanthan, curdlan and pullulan as the most promising polysaccharides

for various industrial sectors attributing levan with a high commercialization potential. Hence there is a growing body of interest in

research associated with levan and its applications. Currently, levan is not only consumed as a functional food but it is also used as

a natural ingredient in commercial personal care products. Recently, halophilic bacterium Halomonas smyrnensis AAD6T

has been

reported as a high-level levan producer extremophile for the first time by our research group3. Further research on the potential

use of levan by Halomonas sp. as a bioflocculating agent, its suitability for peptide and protein based drug nanocarrier systems, its

thin films deposited by laser technologies, levan-based adhesive surfaces obtained by Layer-by-layer (LbL) technology and

comprehensive characterization of its ternary composites were reported. This talk will give a general perspective on the importance

of levan as a biological glue.

References

1- Kang, S.A. , Jang, K.H. , Seo, J.W. , Kim, K.H. , Kim, Y.H. , Rairakhwada, D. , Seo, M.Y. , Lee, J.O. , Ha, S.D. , Kim, C.H., Rhee, S.K. ,

Levan: Applications and Perspectives, Microbial Production of Biopolymers and Polymer Precursors: Applications and Perspectives,

ed: Bernd H.A. Rehm, Caister Academic Pres, (2009). Pp:145-161

2- Combie, J. Properties of Levan and Potential Medical Uses, Polysaccharides for Drug Delivery and Pharmaceutical Applications,

June 22, (2006) Vol.934, 263-269.

3- Poli, A. , Kazak, H. , Gurleyendag, B. , Tommonaro, G. , Pieretti, G. , Oner, E.T. , Nicolaus, B. High Level Synthesis of Levan by a

Novel Halomonas Species Growing on Defined Media , Carbohydr. Pol. , 78 (2009) 651-657.

The financial support provided by Tubitak through projects 111M232 and 112M330 is gratefully acknowledged.

49

Panel 2

Chemical and Structural Characterization

of Natural Adhesives

50

OP-12

INTEGRATION OF SURFACE PLASMON RESONANCE IMAGING IN A TIRF

MICROSCOPE, FOR REAL-TIME, LABEL-FREE IMAGING OF BIOADHESION

PROCESSES

Thomas Ederth, Roni Nugraha, Samira Barhemat, Wetra Yandi

Division of Molecular Physics, IFM, Linköping University, SE-581 83 Linköping, Sweden

Background: Marine organisms use a multitude of strategies, mechanisms and chemistries for attachment to solid surfaces. Efforts

to identify and understand processes involved in temporary and permanent adhesion are motivated by the need for non-toxic

antifouling coatings or exploitation of the excellent underwater performance of marine adhesives in biomedical or technological

applications. Many organisms have planktonic dispersal stages, and good microscopy techniques are essential for studies of their

adhesion.

Methods: Imaging Surface Plasmon Resonance (iSPR) permits real-time, label-free and highly sensitive detection of distributed

surface interaction or adsorption events. We have integrated an iSPR facility into a Total Internal Reflection Fluorescence (TIRF)

microscope, thus greatly expanding the suite of imaging methods available to complement iSPR data.

Results-Discussion: Beyond integration of iSPR with other imaging techniques (phase contrast, darkfield, epifluorescence, TIRF,

etc), this integration of two inherently surface-sensitive methods permits parallell monitoring of exploratory behaviour, imaging of

surface interactions or deposition of material, as well as quantification of deposited amounts, localization and quantification of

adhesive deposits, and subsequent in-situ identification using e.g. fluorescent antibodies, or correlation with ex-

situ characterization using infrared/Raman microscpectroscopy, or imaging XPS.

We demonstrate the capability of this setup using marine model fouling organisms.

Conclusion: This combination of methods will greatly facilitate detailed in-situ studies of the establishment of adhesive joints, and

provide tools for investigating, for example, how temporary adhesives are used to probe surface properties during exploratory

behaviour, or to establish correlations between surface physicochemical properties and quantitative and qualitative properties of

adhesive deposits.

51

OP-13

FAST-TRACK DISCOVERY OF BIOADHESIVES USING A COMBINED HIGH

THROUGHPUT TRANSCRITPTOMICS AND PROTEOMICS APPROACH

Ali Miserez1, Paul Guerette

1, Shawn Hoon

2, Yiqi Seow

2, Clarinda Sutanto

1, Fong T Wong

2

1Nanyang Technological University 2Agency for Science, Technology, and Research (A*Star), Singapore

While many protein-based bioadhesives have been discovered in the past decade, there remains many more to be elucidated.

However, obtaining the primary sequences of uncharacterised adhesives is time-consuming, involving lengthy steps in molecular

cloning and proteomics, thus slowing down new bioadhesive discovery. Therefore, there is an interest to accelerate the discovery

of unknown bioadhesive systems.

Recent efforts to rapidly sequence protein-based adhesives using next generation RNA-seq in conjunction with high-throughput

proteomics will be presented (1). As a proof of concept, the talk will focus on mussel adhesive proteins from the Asian green mussel

(Perna viridis), a major biofouler across the Asia-Pacific region. P. Viridis RNA library of the mussel foot was generated using an

"Illumina GA IIx" system, followed by transcriptome assembly using the "Trinity software". The transcriptome library was then

probed using proteomics data from three complementary methods. First, internal sequences of purified P. Viridis Foot Proteins

(PVFP) were obtained by LC/MS-MS and de novo sequencing. Second, N-terminal sequences was obtained from P. viridis footprint

samples by Edman sequencing. Third, PVFPs of the adhesive plaques were probed by Maldi-Tof.

This high-throughput platform rapidly led to the complete identification of all PVFP adhesive proteins, which showed distinct

molecular features in comparison to the Mytilus system. The approach also allowed to identify tyrosinase enzymes involved in the

post-translational modification of Tyrosine residues into adhesive DOPA.

(1) P. Guerette et al., Accelerating the design of biomimetic materials by integrating RNA-seq with proteomics and materials science, Nature

Biotechnology, 31, 908–915 (2013).

This work is supported by the Singapore National Research Foundation (NRF) and by the Maritime Port Authority (MPA) of

Singapore under the umbrella of the Energy Research Institute at NTU (ERI@N).

52

OP-14 IDENTIFICATION OF NOVEL MARINE PHENOLOXYDASES BY HIGH THROUGHPUT

RNA SEQUENCING

Barbara Maldonado1, Elise Hennebert

2, Patrick Flammang

2, Cecile Van De Weerdt

1

1Molecular Biology and Genetic Engineering Laboratory, GIGA, University of Liège, Liège, Belgium 2Laboratoire de Biologie des Organisms Marines et Biomimétisme, Université de Mons, Mons, Belgium

Marine adhesive proteins are very attractive as potential biomedical and environmentally friendly adhesives thanks to their strong

adhesion to different materials in aqueous media, biocompatibility and biodegradability.

Several adhesive proteins have been identified from the byssus of the blue mussel Mytilus edulis. The adhesive properties are

conferred by post-translational modifications of tyrosine residues into 3,4 dihydroxyphenylalanine (DOPA) by a

monophenoloxydase and the transformation of DOPA residues into DOPA quinone by a diphenoloxydase. However, the enzymes

responsible of these modifications are still unknown.

This study intends to investigate the phenoloxydases responsible of the transformation of tyrosine residues into DOPA and DOPA

quinone in mussels (Mytilus edulis) and tube-worms (Sabellaria alveolata).

To achieve our goals, we performed high throughput RNA sequencing of the adhesive organs of both model organisms. Similarity

search against our two transcriptomes identified five transcripts ofM. edulis and three of S. alveolata that encode proteins

analogous to tyrosinases from a variety of species. Similarity was largely localized in the conserved central domain that contains

two copper-binding sites Cu(A) and Cu(B), the active sites for oxygenase and oxidase function. Expression of the transcripts in their

respective adhesive organ was experimentally confirmed by reverse-transcription PCR (RT-PCR) and sequencing. We will also

determine the localization of these proteins within the cells responsible of the secretion of the adhesive proteins by doing in situ

hybridization (ISH). This will permit to confirm their role in adhesion in M. edulis and S. alveolata.

53

OP-15 COMPARATIVE STUDY OF THE ATTACHMENT OF FUCALES SEAWEEDS FROM

TEMPERATE REGIONS OF THE SOUTHERN HEMISPHERE

Simone Dimartino1, David Savory

2, A. James Mcquillan

2

1Department of Chemical and Process Engineering & Biomolecular Interaction Centre, University of Canterbury, Christchurch, New Zealand 2Department of Chemistry, University of Otago, Dunedin, New Zealand

Background: In this work, Durvillaea antarctica and Hormosira banksii were considered as ideal candidates to investigate the strong

permanent glue produced by macroalgae. In fact, both species are closely related and reproduce sexually. Yet, D. antarctica strives

in wave exposed conditions, while H. banksii only survives in sheltered areas.

Aim: A comparative study of the secretions produced by the two kelp species was conducted to reveal the adhesion mechanism

and the chemical functionalities responsible for seaweed attachment.

Materials-Methods: Attenuated Total Reflection - Fourier Transform Infrared (ATR-FTIR) spectroscopy was used to detect in situ

and real time the chemical functional groups, conformational changes and adsorption reactions involved during seaweed

attachment. Furthermore, the secretion mechanism of the adhesive was studied over time by means of optical microscopy and

Cryo-SEM.

Results-Discussion: Experiments were performed with fresh eggs, sperm and fertilized zygotes while settling onto the ATR active

surface. Physiological conditions similar to that of the natural environment were maintained during the tests, thus allowing for an

appropriate growth of the plants. The IR spectra reveal the production of proteinaceous and carbohydrate based adhesive

molecules.

The microscopic images highlight the production and spread of the adhesive materials, as well as the formation and evolution of

the zygote adhesive pad.

Conclusion: Production of the adhesive components differs between the two seaweed species studied, pointing out differences in

the adhesion chemistry and mechanism. A new model on the formulation and structure of the glue produced by Fucales seaweed is

also proposed.

54

OP-16 INVESTIGATION OF THE ADHESION OF LEVAN VIA MOLECULAR DYNAMICS

SIMULATIONS

Gulcin Cem1, Binnaz Coskunkan

1, Seyda Bucak

1, Deniz Turgut

2, Deniz Rende

2, Nihat Baysal, Rahmi Ozisik

2,

K. Yalcin Arga3, Ebru Toksoy Oner

3,

1Department of Chemical Engineering, Yeditepe University, Istanbul 34755, Turkey 2Department of Materials Science and Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA 3Department of Bioengineering, Marmara University, Istanbul 34722, Turkey

Background: Levan is a biopolymer consisting of β–D–fructofuranose units with β(2–6) linkages between fructose rings. Preliminary

experimental studies showed that levan has excellent adhesive properties, which would present various opportunities in many

applications such as wound healing. Understanding of the adhesive properties at the molecular scale could provide further

opportunities to improve and tailor the properties of levan and even target applications in nanoelectronics.

Molecular Dynamics (MD) simulations were used to investigate the static, dynamic, and adhesive properties of levan chains on

graphene. Graphene surface chemistry was changed to mimic experimental conditions.

55

OP-17 THE PROJECTILE SLIME OF VELVET WORMS – STRUCTURAL CHARACTERIZATION OF

THE BIOLOGICAL ADHESIVE IN ONYCHOPHORA

Alexander Baer1, Matthew J. Harrington

2, Stephan Schmidt

3, Georg Mayer

1

1Animal Evolution and Development, Institute of Biology, University of Leipzig, Talstraße 33, D-04103 Leipzig, Germany 2Dept. of Biomaterials, Max Planck Institute of Colloids and Interfaces, Am Mühlenberg 1, D-14476 Potsdam, Germany 3Biophysical Chemistry, Institute for Biochemistry, University of Leipzig, Johannisallee 21-23, D-04103 Leipzig, Germany

Background-Aims: Onychophorans use an adhesive, protein-based slime for prey capture and defence, which is ejected via a pair of

modified limbs to immobilize their prey. The slime shows four major characteristics as potent biological adhesive, (i) fast wetting (ii)

universal adhesiveness (even under water); (iii) mechanical locking; and (iv) high tensile strength after curing. To gain further

insights into the chemical and structural mechanisms underlying these properties, we analyzed the composition and architecture of

the slime and adhesive fibers of the onychophoran Euperipatoides rowelli at the microscale, nanoscale, and molecular level.

Results-Discussion: Our results at the microscale level indicate that the crude slime is an unstructured mix of proteins. However,

after mechanical agitation, the slime forms sticky solid fibers. Importantly, as observed by optical and polarized light microscopy,

fibers consist of a core region comprised of oriented fibrils with thicknesses on the order of a few micrometers and a thin adhesive

coating that is apparently unstructured. In spite of the fibrous appearance of the core, orientation is not evident at the nano- and

molecular scale as determined by X-ray diffraction and polarized Raman spectroscopy methods. On the other hand, DLS and AFM

imaging in the sub-micron regime revealed that thread-forming proteins in the crude slime may possibly exist as micelle-like

globules with low size distribution (100–500 nm) that form micro-fibrous aggregates after experiencing mechanical stress.

Conclusion: These tentative findings on the onychophoran slime suggest a novel mechanism that achieves a protein-based bio-

adhesive with surprisingly high performance.

Australian onychophoran species Euperipatoides rowelli

56

OP-18

INVESTIGATIONS ON THE PROTEIN FRACTION OF CUVIERIAN TUBULE ADHESIVE

MATERIAL

M. Demeuldre1, R. Wattiez

2, P. Ladurner

3, E. Hennebert

1, P. Flammang

1

1Biology of marine organisms and Biomimetics, University of Mons, Mons, Belgium 2Proteomics and Microbiology, University of Mons, Mons, Belgium 3Institute of Zoology, University of Innsbruck, Austria

Background: Cuvierian tubules are white caeca used as a defence system in some species of sea cucumbers, all belonging to the

family Holothuriidae. When attacked by a predator, the animal expels tubules which elongate and become sticky upon contact with

a surface. The adhesive material produced by Cuvierian tubules is made up of an inorganic fraction accounting for 10 % of the dry

weight and an organic fraction composed by 54 % proteins and 36 % carbohydrates. It has been proved that following a trypsin

treatment (cutting proteins into peptides), the adhesiveness of the tubules is decreased.

Materials-Methods: On the one hand, proteins were extracted from the adhesive material produced by the tubules. Mass

spectrometry analyses were performed on theses proteins. On the other hand, mRNAs were extracted from whole tubules and an

Illumina transcriptome was generated.

Results: Mass spectrometry analyses yielded a list of all the proteins detected in the tubule adhesive material. Some of these

proteins were identified in the databases and correspond to intracellular proteins (presumably contaminants in our case). Some

other proteins were not identified and are therefore particularly interesting, especially if they are quite abundant in the adhesive

material. Peptides corresponding to these unknown proteins were blasted against the transcriptome to obtain the complete

primary sequence of these proteins presumably involved in adhesion.

Discussion: After the elaboration of a list of candidate proteins, putative adhesive proteins must be tested by in situ hybridization

to localize their expression in the tubule and therefore confirm their function.

57

Panel 3

Molecular Mechanisms of Bio-Adhesion

58

OP-19 CLONING, EXPRESSION AND PURIFICATION OF NECTIN, A SEA URCHIN ADHESIVE

PROTEIN

Duarte Toubarro1, Analuce Gouveia

1, Catarina Alcarva

2, Raquel Mesquita Ribeiro

2, Nelson Simões

1, Gonçalo Conde Da Costa

2,

Carlos Cordeiro2, Romana Santos

3

1Centro Investigação Recursos Naturais do Centro de Biotecnologia dos Açores, Departamento de Biologia, Universidade dos Açores, Ponta Delgada,

Portugal 2Centro de Química e Bioquímica, Departamento de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa, Lisboa, Portugal 3Unidade de Investigação em Ciências Orais e Biomédicas, Faculdade de Medicina Dentária da Universidade de Lisboa, Portugal

Background: The sea urchin Paracentrotus lividus is a common inhabitant of wave-swept shores relying on adhesive organs, tube

feet, to produce an adhesive secretion, with which they attach, strongly but temporarily, to the substrate. The adhesive

composition is still poorly known, but recently, the proteome of feet discs (that enclose the adhesive precursors), pinpointed some

adhesive proteins, such as Nectin – known to be involved in P. lividus embryo’s cell/substrate adhesion.

Materials-Methods: To access the adhesive potential, Nectin cDNA was obtained from tube feet discs, cloned and sequenced. The

full-length Nectin cDNA (2950bp) comprising 6 galactose-binding domains and a fragment containing only 4 domains (2019bp) were

produced, inserted into pGEX fusion vector, expressed in E. coli (C41 and R2) and purified by affinity-chromatography. Expression

levels of Nectin were also evaluated by RT-PCR in different tissues.

Results: The Nectin gene sequence obtained from feet discs has 90% of identity with the sequence annotated for embryo but with

15 mutated amino acids. This Nectin gene variant seems to be specifically expressed in discs, whereas other tissues preserved the

variant expressed in the embryo. Genome analysis reveals traces of at least one gene duplication event. Nectin expression and

purification is being optimized and the recombinant protein tested for its adhesiveness.

Conclusion: We provide the first evidences that P. lividus adults are also using the adhesive abilities of Nectin for substrate

attachment, although through a new variant of this protein.

59

OP-20 HOMOLOGY BUT LOW SIMILARITY IN ADHESIVE PROTEINS OF STALKED AND

ACORN BARNACLES

Anne Marie Power1, Jaimie Leigh Jonker

1, Florence Abram

1, Ana Varela Coelho

2, Ingo Grunwald

3

1School of Natural Sciences, National University of Ireland, Galway, Ireland 2Instituto de Tecnologia Química e Biológica, New University of Lisbon, Av. da República, Estação Agronómica Nacional, 2780-157 Oeiras, Portugal 3Fraunhofer Institute for Advanced Biomaterials Research, Wiener Str. 12, 28359 Bremen, Germany

Background-Aim: Information about a range of species within the barnacles may be instructive to identify conserved adhesive

domains. Our aim was to add to extensive information from the acorn barnacles (order Sessilia) by providing the first protein

analysis of the adhesive of L. anatifera (order Lepadiformes).

Materials-Methods: The proteins of L. anatifera were investigated using 1D SDS-PAGE and mass spectrometry. Polyclonal

antibodies isolated from acorn barnacle species were tested on L. anatifera adhesive glands and gene similarities across barnacle

groups were examined using alignments.

Results (quantitative and / or statistical data): 12 protein bands were evident after cement solubilisation in L. anatifera, with

intense bands at ~30, 70, 90 and 110 kD. 78 peptides of 7-16 amino acids in length were sequenced de novo from these bands.

None of the peptides matched published or unpublished transcriptome sequences, but there was some similarity between L.

anatifera and closely-related Dosima fascicularis. Antibodies from two acorn barnacle proteins (ab-cp-52k and ab-cp-68k) showed

cross-reactivity in the adhesive glands of L. anatifera. Sequence alignment clearly showed homologues in stalked barnacles for the

19 kD and 100 kD proteins in acorn barnacles.

Discussion: Several of the proteins described in acorn barnacles appear to have homologous proteins in stalked barnacles; cp-19k

and cp-100k are also found in P. pollicipes, while cp-52k and cp-68k appear to be expressed in L. anatifera adhesive tissues.

However whilst there is homology, sequence similarity in amino acid and gene sequences tended to decline as taxonomic distance

increased.

60

OP-21 TOUGH COATING PROTEINS: SUBTLE SEQUENCE VARIATION MODULATES

COHESION

Saurabh Das1, Dusty R Miller

2, Yair Kaufman

1, Maryte Gylys

3, Jacob N Israelachvili

4, Herbert J Waite

5

1Department of Chemical Engineering, University of California, Santa Barbara, California 93106, USA 2Biomolecular Science and Engineering, University of California, Santa Barbara, California 93106, USA 3Department of Chemistry and Biochemistry, University of California, Santa Barbara, California 93106, USA 4Department of Chemical Engineering; Materials Research Laboratory, University of California, Santa Barbara, California 93106, USA 5Biomolecular Science and Engineering; Department of Chemistry and Biochemistry; Materials Research Laboratory, University of California, Santa

Barbara, California 93106, USA

Mussel foot protein-1(mfp-1) is a coating protein that forms the major constituent of the protective cuticle covering all exposed

portions of the byssus that anchors the marine mussels to mineral surfaces in harsh intertidal zone. The reversible interaction

between the DOPA moieties in the protein and Fe3+ in the cuticle is thought to be responsible for its ability to accommodate core

strains of up to 120% in M. californianus. We have investigated the cohesive interactions between mcfp-1 films deposited on mica

surface at various bulk protein deposition concentrations. Our results show that the cohesive force of interaction between the

organic films of mcfp-1 is maximum for protein deposited at 50 μg/ml. Increasing the deposition concentration decreased the

interaction energy significantly showing that increasing the amount of adhesive does not always increase adhesion. Addition of

Fe3+ did not change the cohesion energy between the mcfp-1 films significantly at pH 5.5. However, iron mediates the bridging

between the mcfp-1 films at the physiologically relevant pH of 7.5, when most of the mussel foot proteins lose their ability to

adhere reversibly. This preservation of cohesion between the protein films at high pH is through the formation of multivalent

bonds between Fe3+ and DOPA which prevents the oxidation of DOPA to DOPA-quinone. Considering the cuticle of M.

Californianus can withstand twice the strain of those in M. edulis bolsters the argument that the innate attraction found in mcfp-1

films may be important for the engineering of strain tolerant composite coatings for biomedical applications.

61

OP-22 CATION-PI INTERACTION: NEW INSIGHT FOR UNDERWATER ADHESION

Dong Soo Hwang

School of environmental science and engineering, Pohang university of science and technology, Pohang, South Korea

Background-Aim: Cation-pi interactions are one of the most important noncovalent interactions in biological systems. cation-pi

interaction that is widely present between an electron-rich pi system (e.g., Phe, Tyr, Trp) and adjacent cations (e.g., Na+, K+) or

species containing positive charges (e.g., Lys, Arg) in the physiological conditions of the living organisms

Recent studies on adhesion mechanism of mussel adhesive proteins by using an Surface forces apparatus (SFA) suggest Cation-pi

interaction as an important underwater adhesion mechanism.

Materials-Methods: In this report, the mechanical nature of cation-pi interactions in aqueous media was probed directly using a

surface forces apparatus (SFA) for the first time.

Results (quantitative and / or statistical data): Strong and reversible underwater cohesions between two opposing mfp-1 films

which do not have any positively charged amino acid residues in testing pH were measured even with passivation of DOPA. Cation-

pi interaction, which act between an electron-rich π system (egs. Tyr, Dopa, Phe, Trp) and adjacent cations (egs.,n-terminus amine,

Lys, Arg, His), was suggested as a main contributor to the cohesive nature of two opposed mfp-1 layers. SFA measurements and

theoretical simulations also confirm that cation-pi interactionsin aqueous media depend on the type and concentration of

positively charged ions (Lys+, NH4+, K+, Li+, Na+) and type of aromatic side groups (indole, phenol, benzene) present in solution.

Discussion: Cation-pi interaction is a noncovalent bonding which is comparable in strength to hydrogen bonding and electrostatic

bond in aqueous solutions.

Conclusion: Cation-pi interaction is a key contributor for underwater adhesion.

Aqueous cation-pi interaction schemes

62

OP-23 A NEW NUMERICAL CODE FOR HIERARCHICAL PEELING SIMULATIONS

Lucas Brely1, Federico Bosia

1, Nicola M Pugno

2

1Department of Physics, University of Torino, Italy 2Department of Civil, Environmental and Mechanical Engineering, Università di Trento, Italy

Background-Aim: Adhesion of geckos or other insects to surfaces has been widely studied in recent years, including their ability to

adapt to both rough and smooth surfaces, achieve smart detachment through control of contact angles, and activate self cleaning.

Bioinspired synthetic fibrillar dry adhesives have been developed mimicking these biological structures, but much remains to be

done in the optimization their performance.

To achieve a better understanding of adhesion mechanisms and how they can be optimized, we developed a novel numerical code

to simulate adhesion and peeling of multiple fibrillar structures of arbitrary geometry, including hierarchical architectures.

Materials-Methods: Results (quantitative and / or statistical data): We first validated the numerical code by comparing results with

known results using the theory of multiple peeling, which extends the energy-based single peeling theory of Kendall, including large

deformations and pre-stretching. Then, we we studied the adhesion of model structures with single and multiple contacts, in

simple and increasingly complex geometries, with the aim of determining optimal configurations.

Discussion-Conclusion: Model validation provides good confidence in the predictivity of the code. The influence of complex

configurations is discussed and the beneficial effects of hierarchical structures demonstrated.

63

OP-24 WHEN THE GOING GETS ROUGH... THE EFFECT OF SURFACE ROUGHNESS ON THE

ATTACHMENT ABILITIES OF TREE FROGS

Niall Crawford, Thomas Endlein, Mathis Riehle, W. Jon. P. Barnes

Centre for Cell Engineering, Institute of Molecular, Cell and Systems Biology, University of Glasgow, Glasgow, U.K.

Background: Tree frogs use specialised toe pads to stick using wet adhesion, producing strong adhesive and friction forces when

tested on smooth surfaces. However, how do frogs cope with surfaces of varying roughness they would naturally encounter?

Methods: The attachment performance of tree frogs (Litoria caerulea) against rough surfaces was tested by measuring 1) the slip

and fall angles of whole animal on a tilting apparatus; 1) single toe pad forces using a custom-built 2-D force transducer; and 3) the

conformation of the soft pad tissue against asperities using interference reflection microscopy (IRM).

Results: Small scale roughness (0.5 to 30 microns) produced optimum friction in the whole animal experiments. On rougher

surfaces (30 to 325 microns), the frogs struggled to stick and subsequently slipped and fell off at lower angles. Measurements on

single toe pads confirmed these results. Preliminary data obtained using IRM showed that pads conform closely around asperities

smaller than 50 microns, with the adhesive fluid filling gaps.

Discussion: Similar to insects and geckos, tree frogs show varying performance on different length scales of roughness. We believe

that parameters like the pad’s softness to conform, the resistance to interlock and the ability of the fluid to fill cavities create a

complex pattern of attachment performance.

Conclusion: By using soft toe pads and an adhesive fluid, tree frogs can adhere even to rough surfaces. Surprisingly, they struggle to

stick to certain rough surfaces.

Understanding the mechanism behind this behaviour would benefit the development of 'smart' adhesives.

64

OP-25 WET BIOADHESION IN TREE FROGS

Farzaneh Kaveh1, Ciro Semprebon

2, Michael Kappl

1, Hans Jürgen Butt

1

1Max Planck Institue for Polymer Research 2Max Planck Institute for Dynamic and Self-Organization

Background-Aim: In nature, many animals are able to attach to surfaces by using their special adhesive pads such as geckos and

tree frogs. By mimicking the reversible adhesion in animals we could benefit it in our daily life by using reversible plasters and or

having tires with better traction in wet conditions. Tree frogs are able to adhere strongly and release easily from surfaces even in

wet conditions. The hexagonally patterned structure on their toe pads help tree frogs to have strong reversible adhesion.

Materials-Methods: We have fabricated soft, elastic PDMS microstructures consisting of hexagonal pillars that mimic these toe

pads. Pillar structures with different terminal shapes and hydrophilic or hydrophobic surface were studied. Adhesion forces for

single pillars were measured by atomic force microscope using specially designed colloidal probes.

Results (quantitative and / or statistical data): To separate the different contributions of surface forces, we have carried out

experiments in presence and absence of an intervening liquid layer and at different separation speeds. The system was simulated

too.

Discussion: Our experiments approved that hydrodynamic force does not play a role in wet adhesion in case of walking in tree

frogs. Good agreement have seen between experimental and simulation curves explained by capillary theory.

Conclusion: Adhesion force under dry condition is higher on T-shape pillar than on Normal shape and least on concave cups. In a

wet condition, the adhesion force depends on the amount of liquid that wets the surface.

Figure

Direct comparison between experimental and simulation curve.

65

OP-26 ONE SOLUTION FOR ALL CASES: ADHESIVE ABILITY OF THE LEAF BEETLE’S FEET

Naoe Hosoda1, Stanislav N Gorb

2

1Hybrid Materials Unit, National Institute for Materials Science, Namiki 1-1, Tsukuba, Ibaraki, Japan 2Zoological Institute of the University of Kiel, Am Botanischen Garten 1–9, Kiel, Germany

Background-Aim: A design which enables easy disassembly is an important requirement for environment- friendly products, and

reversible interconnection techniques are the key technology for this purpose. The aim of our research is development of reversible

interconnection by biomimetic approaches. We investigated adhesive ability of the feet of the leaf beetle Gastrophisa viridula as a

model species. The effects of substrate topography and wettability on the adhesive ability were investigated. Furthermore, we

investigated the adhesive ability of beetle’s feet underwater.

Materials-Methods: Beetles G. viridula were collected from their host plant Rumex obtusifolius. Various substrates with different

wettability and different surface roughness were prepared. The substrate roughness was described and quantified as height (rms)

and curvature of surface asperities. Individual beetles were connected to the force sensor and traction force generated by each

beetle was measured in the air and underwater.

Results-Discussion: The adhesive ability of the beetle’s feet was strongly reduced at substrate roughness of about 100 nm (rms)

due to minimization of the contact area between single hairs of the foot and substrate. The leaf beetle was less influenced by the

surface wettability in the air. For the first time, we revealed and experimentally studied adhesion ability of the beetle using air

bubbles underwater.

This work was supported by JSPS Grant-in-Aid for Scientific Research on Innovative Areas Grant Number 24120005.

66

OP-27 STICKY UNDERWATER CADDISFLY SILK: ELASTIC, METAL-ION DEPENDENT, DOUBLE

NETWORK FIBERS

Russell J. Stewart, Nicholas N. Ashton

Department of Bioengineering, University of Utah, Salt Lake City, Utah, USA

Background: Caddisflies are aquatic insects related to terrestrial moths and butterflies. Like their terrestrial cousins, aquatic

caddisfly larvae spin silk from a pair of modified salivary glands. The adhesive silk is used to assemble a variety of structures

underwater, including composite body armor and food harvesting nets. Caddisfly H-fibroins contain (pSX)n motifs, where pS is

phosphoserine and n = 2-6.

Methods: Silk fiber molecular structure and mechanical properties were correlated using a combination of ATR-FTIR and single

fiber micromechanical testing. Peroxidase and dityrosine crosslinks were identified in silk fibers by LC-MS. A peroxidase gene was

identified in the assembled silk gland transcriptome.

Results: Native fibers contained a 0.9:1 Ca to P ratio, and much smaller ratios of other metal ions. Mechanically, single native fibers

are highly extensible, display a distinct yield point, force plateau, and load cycle hysteresis. Metal ion exchange with Na EDTA

decreased β-sheet content, shifted the FTIR phosphate peaks from a Ca2+-complexed form to uncomplexed, and decreased the

initial modulus, toughness, and failure stress to ~1% of native fibers. Re-addition of Ca2+ restored the β-sheet content, complexed

phosphate, initial modulus, and load cycle hysteresis. Exchange with other metal ions had pronounced effects on mechanical

properties. The sigmodal pH dependence of the mechanical properties superimposed the FTIR pH titration curve of the phosphates.

Conclusions: (pSX)n motifs may form Ca2+-stabilized β-domains functionally analogous to β-domains of moth silks, and impart

strength and energy-dissipating qualities to underwater caddisfly silk fibers. Peroxidase catalyzed dityrosine crosslinks create a

weak elastic second network.

67

Panel 4

Bioadhesion: Prevention

68

OP-28 WHERE ADHESION FAILS: SLIPPERY SURFACES OF INSECT-TRAPPING PITCHER

PLANTS

Ulrike Bauer1, Holger Florian Bohn

2, Walter Federle

3

1School of Biological Sciences, University of Bristol, Bristol, UK 2Plant Biomechanics Group Freiburg, Botanic Garden, Faculty of Biology, University of Freiburg, Freiburg, Germany 3Department of Zoology, University of Cambridge, Cambridge, UK

Background-Aim: Insects affect human life and economy more than any other animal class. While the diligence of pollinators is

essential for our food supply, disease-transmitting insects and agricultural pests pose serious challenges. Some plants have evolved

amazingly effective insect-repellent surfaces that have the potential to inspire the bio-engineering of physically protected crop

plants or the design of anti-adhesive surface coatings.

Materials-Methods: We performed running experiments and friction force measurements with ants to characterize the function

and properties of the slippery trap surfaces of carnivorous pitcher plants.

Results (quantitative and / or statistical data): Unrelated Asian Nepenthes and South-American Heliamphora pitcher plants have

evolved strikingly similar mechanisms despite a radically different surface structure. The corrugated upper pitcher rim

of Nepenthes and the hairy inner wall of Heliamphora pitchers are both highly wettable. Water spreads and forms thin films on

these surfaces, rendering them extremely slippery for insects. In contrast, Nepenthes gracilis has a uniquely 'fine-tuned' micro-

rough wax crystal surface which provides access to nectar-foraging ants but is slippery enough for them to become dislodged by

the impact of rain drops.

Discussion: Our results show that pitcher plant trapping mechanisms are more diverse that previously assumed. Pitcher plants

employ two fundamentally different anti-adhesive mechanisms: 'insect aquaplaning' on superhydrophilic, water-lubricated

surfaces, and contact area reducing surface micro-roughness. Functional convergence in unrelated taxa has the potential to

highlight key properties of anti-adhesive plant surfaces.

Conclusion: Pitcher plants constitute an ideal model system to study natural anti-adhesive surfaces.

69

Figure 1

Unrelated Nepenthes (A-C) and Heliamphora (D-E) pitcher plants have evolved strikingly similar mechanisms to trap insects. The

upper rim (peristome) of Nepenthes pitchers (A) is readily wettable (B) due to its specialized surface microstructure (C). The inner

wall of Heliamphora pitchers (D) is covered with a dense carpet of hydrophilic trichomes (E). Both surfaces become extremely

slippery when wet. (Scale bars: 100µm)

Figure 2

Nepenthes gracilis has evolved an 'inverted springboard' trapping mechanism. The underside of the pitcher lid is covered with a

glaucous wax crystal layer (A, C) and highly productive nectaries (B) to attract insects. The unique pillar-like microstructure (D-E)

provides enough grip for insects to access the nectar but reduces adhesion sufficiently for them to become dislodged by the impact

of rain drops. (Scale bars: 5 µm)

70

OP-29 STUDY THE INTERACTION OF CYPRID TEMPORARY ADHESIVE PROTEINS AND

SURFACES USING AFM

Shifeng Guo1, Dominik Janczewski

1, Reddy Sreenivasa Puniredd

1, Xiaoying Zhu

1, Serina Siew Chen Lee

2, Serena Lay Ming Teo

2,

Julius Gyula Vancso3

1Institute of materials research and engineering A*STAR (Agency for Science, Technology and Research), 3 research link Singapore 117602 2Tropical marine science institute, National University of Singapore, 18 Kent ridge road Singapore 119227 3Institute of chemical and engineering sciences A*STAR, 1, pesek road, jurong island, Singapore 627833; MESA+ institute for nanotechnology,

materials science and technology of polymers, University of Twente, P.O. box 217, 7500 AE Enschede, The Netherlands

Background-Aim: Cyprid footprint proteins (FPs), which are temporary protein adhesives secreted when the larvae explored

surfaces, play significant role on the barnacle induced marine biofouling.

Materials-Methods: Atomic force microscopy (AFM) was used to study the interaction of the temporary adhesive proteins and

surfaces of different wettability and charge. The FP proteins were immobilized on an AFM colloidal probe and used as a force probe

to study the interaction of proteins with the various surfaces based on force spectroscopy method.

Results (quantitative and / or statistical data): The morphology of FPs was characterized in seawater condition and showed

significant larger sizes on the hydrophobic surfaces. However, the overall volume of proteins secreted on the surfaces showed no

significant. The FP proteins had a stronger interaction with the hydrophobic surface of 21± 2 nN, which is much larger than the

value measured on hydrophilic surface of 7.2 ± 1 nN. Also FP proteins showed greatest adhesion on the anionic surfaces than the

neutral and cationic surfaces.

Discussion-Conclusion: Cyprid FP proteins undergo conformational change when interact with surfaces of different wettability and

adhere more strongly on the hydrophobic surfaces. Also, the proteins are verified to be positively charged and showed stronger

adhesion with the negatively charged surfaces. Combined with the settlement data, it may be concluded that surface charge, other

than the surface wettability, should be more considered on the design of antifouling surfaces.

The authors are grateful to the Agency for Science, Technology and Research (A*STAR) for providing financial support under the

Innovative Marine Antifouling Solutions (IMAS) program.

71

Panel 5

Catechol-based Adhesives

72

OP-30 MUSSEL-INSPIRED ADHESIVE INTERFACES FOR BIOMEDICAL APPLICATIONS

Hakan Ceylan, Samet Kocabey, Hilal Unal Gulsuner, Ayse Begum Tekinay, Mustafa Ozgur Guler

Bilkent University, National Nanotechnology Research Center

Engineering cell-biomaterial interface with instructive coatings have become an appealing strategy to stimulate tissue regeneration

and improve material biocompatibility. Guiding cellular activities, such as adhesion, proliferation, and differentiation, at this

interface is often essential for both accelerated healing and the long term clinical success of the surgical intervention. In the

biological milieu, however, stability of coatings on the material surface is limited under highly abrasive biological conditions.

Analogously, various organisms, such as mussels, adapted to living in the seashores, suffer from the harsh physical and chemical

instabilities. To remain sessile, mussels synthesize a highly complex, spatiotemporally evolving glue containing high amount of 3,4-

dihydroxy-L-phenylalanine (DOPA). Herein, we imitate this mechanism by incorporating it to self-assembling peptide amphiphile

building blocks. Self-assembly of these molecules with other peptide amphiphiles carrying biofunctional ligands, such as REDV,

DGEA, and KRSR, form nanofibrous adhesive networks, which closely mimic the structure and biochemical properties of the native

extracellular matrix. We demonstrate the general applicability of this supramolecular bioadhesive by applying as bioactive

cardiovascular stent coating, where endothelial cell adhesion and survival is favored over the smooth muscle cells, which is the

main cause of re-closure of the arteries (restenosis). We further develop an orthopedic/dental implant coating where mineral-

depositing cells are selectively promoted over soft tissue forming fibroblasts. Mussel-mimetic nanofibers can deposit bone-like

hydroxyapatite in body fluid mimetic conditions. This paves the way to develop a second generation orthopedic/dental implant,

which efficiently mediates differentiation of human mesenchymal stem cells into functional osteoblasts.

Mussel-inspired surface biofunctionalization with peptide nanofibers

73

OP-31 CROSSLINKING KINETICS OF CATECHOLAMINE DERIVATIVES

Julieta Paez, Cristina Serrano, Aránzazu Del Campo

Max-Planck-Institut für Polymerforschung. Ackermannweg 10, 55128, Mainz, Germany

Mussels secrete DOPA-containing adhesive proteins that play an important role in their underwater adhesion properties. Aiming at

mimicking mussel performance, cathecol based systems have been the subject of intensive studies. The chemical environment of

the catechol group is known to play an important role in the crosslinking of DOPA modified materials. Here, we report on the

preparation of adhesive precursors based on a branched PEG core derivatized with different catecholamine end groups (DOPA,

norepinephrine, dopamine, Cl-dopamine and NO2-dopamine). These polymers crosslink and form stable gels in oxidative

conditions. The effect of the catecholamine chemical environment on the oxidation, crosslinking degree and gelation kinetics was

studied by rheology, NMR, and UV-Vis spectroscopy. We show that these systems are promising as surgical adhesives.

74

OP-32 FUNCTIONAL HYBRID SURFACES AND INTERFACES BASED ON BIOINSPIRED

CATECHOLS

Daniel Ruiz Molina1, Javier Saiz Poseu

1, Beatriz Garcia

2, Josep Sedo

1, Jordi Hernando

3, Felix Busque

3

1Institut Catala de Nanociènica i Nanotecnologia, Campus UAB, 08193, Cerdanyola del Valles,Spain 2Fundación Privada ASCAMM, Unidad de Nanotecnología, ParcTecnològic del Vallès, Av. UniversitatAutònoma, 23 - 08290 Cerdanyola del Vallès,

Spain 3Chemistry Department, Universitat Autònoma de Barcelona, Campus UAB 08193, Cerdanyola del Vallès, Spain

An ever-increasing number of bioinspired catechol-based polymers (mainly polydopamine) have been reported and shown to

constitute powerful tools for the fabrication of hybrid materials as water-resistant adhesives, protective layers, and primers for

functional adlayers and nanoscale coatings, among others. [1]

Herein we report a new synthetic approach for the fabrication of such catechol-based materials with adherent properties. The

mechanism is based on a simple polymerization process in the presence of ammonia [2], in a way reminiscent of melanization

reactions. The initial molecule of choice was a catechol bearing a long alkyl chain. The resulting material after polymerization,

spontaneously self-assembles in the form of nanoparticles that easily stick to different surfaces and materials. As an application,

the NPs have been successfully implemented for water remediation of heavy metal ions and are being explored nowadays for drug

delivery.

On the other side, when this material is placed in non-polar solvents such as hexane, the NPs are dissolved and the polymer is used

to obtain coatings thanks to their adhesive properties. Coatings are effective on a representative variety of substrates, both at the

nano-/macroscale, without any pretreatment or interphase modification. Whereas a long alkyl chain affords coatings with a

persistent hydrophobic character, this methodology can be extended to several other catechols with different ring pendant groups,

providing additional surface functionalities such as oleophobic, anti-fouling or anti-bacterial activities [3].

[1] Adv. Mater. 2013, 25, 653.

[2] Adv. Mater. 2013, 25, 2066.

[3] Small 2014, DOI: 10.1002/smll.201302406

Multifunctional catechol-based hybrid materials

75

Panel 6

Industrial Challenges in Adhesion

76

OP-33 NATURE OF THE PROBLEMS RELATED TO E. COLI ADHESION ON UNIFORM FILMS

PREPARED AT LOW TEMPERATURE HINDERING BACTERIAL INACTIVATION:

CRITICAL ISSUES

John Kiwi

S. Rtimi, C. Pulgarin, J. Kiwi, Ecole Polytechnique Fédérale de Lausanne(EPFL), Station 6,CH 1015, Lausanne, Switzerland

Background-Aim: Preparation of supported semiconductor-metal uniform films at low temperatures showing fast inactivation

kinetics(minute range), mechanistic aspects and practical hindrance of biofilm formation and airborne bacterial density

Materials-Methods: sputtering of substrates below 120 oC deposits semiconductor-metal films leading to bacterial inactivation in

the dark and under light. Bacterial inactivation kinetics and surface properties has been evaluated for the nitrides, metal-nitrides

and TiO2/Cu films.

Results-Discussion: Due to the poor adhesion of metals to low thermal resistant substrates prepared from colloids, we have used

sputtering techniques in the magnetron chamber (100-150 oC) deposit uniform metal films. TiO2 photocatalyzed bacterial

inactivation proceeds with slow kinetics absorbing only 4-5% of the solar spectrum. Our innovative film composites consist of

metal-semiconductors increase: a) the bacterial inactivation kinetics due to the formation of metal-semiconductor surface layers,

b) the absorption of the TiO2 into the visible region and c) the metal particles adhesion on the substrates due to the added TiO2

interfacial layers. We have recently reported antibacterial films like: TiON, TiON-Ag(1-2), and TiO2/Cu(2), the surface properties and

bacterial inactivation kinetics.

Conclusion: Bacterial inactivation kinetics proceeds in the dark mediated by the composite films and is accelerated under light

(minute range) precluding biofilm formation on low thermal resistant surfaces.

References

(1) Sami Rtimi, Oualid. Baghriche, Rosendo Sanjines, Cesar Pulgarin, Michael Ben-Simon, Jean-Claude Lavanchy, John. Kiwi, J. Photochem. Photobiol.

A, 2013, 256, 52-63.

(2) Sami Rtimi, Oualid Baghriche, Cesar Pulgarin, Jean-Claude Lavanchy, John Kiwi, Surf. & Coat. Technol. 2013, 232, 804-813.

77

OP-34 ADDITION OF TUNG OIL DIOL FOR FACILITATING THE DEGRADATION OF

POLYURETHANE ADHESIVES

Jose Miguel Martin Martinez1, Pilar Carbonell Blasco

2

1Adhesion and Adhesives Laboratory,University of Alicante, Alicante, Spain 2Bioadhesives Medtech Solution (Adhbio), Elche, Spain

Background-Aim: Thermoplastic polyurethane adhesives have segmented structure due to the incompatibility between the hard

and the soft segments. Their phase morphology is determined by the raw materials used in polyurethane synthesis. Current

polyurethane adhesives are not biodegradable neither recyclable. In this study, it is proposed the addition of diol of tung oil as part

of the polyol or the chain extender for facilitating degradation of the polyurethane adhesives.

Materials-Methods: The prepolymer method was used in the synthesis of the polyurethane adhesives. NCO/OH ratio was 1.05, and

the reactants were MDI (4,4´-difenilmethanediisocyanate), polyadipate of 1,4-butanediol (Mw=2500), 1,4-butanediol, and tung oil

diol (Figure 1).

Results-Discussion: Addition of tung oil diol in the chain extender (25BD+75 TO) increases the cross-over between the elastic and

viscous modulus indicating the creation of new hard segments and increases degree of phase separation in the polyurethane.

However, in the polyurethane with tung oil diol in the polyol (85PAD+15TO) the moduli cross-over is not produced.

Thermoplastic polyurethanes were thermally degraded (80°C-95% relative humidity-7d) and their properties were studied. The

degradation of the polyurethane without tung oil increases the storage modulus while it is decreases in the polyurethanes

prepared with tung oil diol, in a greater extent when incorporated in the chain extender. Thermal degradation is produced in the

hard segments (Figure 2).

Conclusion: Addition of tung oil diol facilitates the degradation of polyurethane adhesives acting on the hard segments.

78

Figure 1

Scheme of the polyurethane adhesives synthesis containing tung oil diol in the poliol and in the chain extender.

Figure 2

Variation the storage (G´) and viscous (G’’) modulus as a function of the temperature for the polyurethanes with and without tung

oil diol after degradation.

Financial support by Spanish Research Agency MICINN (MAT2010-19904 project) is acknowledged.

79

OP-35 CROSS-LINKABLE LEVAN DERIVATIVES FOR BIODEGRADABLE ADHESIVES

Ralf Wyrwa1, Albrecht Berg

1, Ebru Toksoy Öner

2, Matthias Schnabelrauch

1

1INNOVENT e. V., Biomaterials Department, Jena, Germany 2Marmara University, Department of Bioengineering, Istanbul, Turkey

Background-Aim: Cross-linkable biomaterials have been successfully applied in different medical applications. Especially

(meth)acrylates are widely used in medicine, e. g. in dentistry or orthopaedic surgery. Recently cross-linkable biomaterials based on

polysaccharides and proteins have been intensively studied for their applicability in tissue engineering applications as well as tissue

adhesives. Regarding medical adhesives, there are urgent needs in trauma and reconstructive surgery. In spite of the high adhesion

strengths of (meth)acrylate based adhesives their inherent bioinert nature often affects biodegradation and consequently the

required replacement by endogenous tissue. Therefore our research is focused on the development of biodegradable adhesives

based on cross-linkable biopolymers including polysaccharides, glycosaminoglycans and proteins usable in tissue engineering

applications especially soft tissue repair.

Materials-Methods: Levan is a high molecular weight fructose-based biotechnologically available polysaccharide with a range of

interesting properties qualifying this molecule for applications in biomedicine. New levan derivatives containing methacrylate

groups attached by different chemical linkages were synthesized and characterized by conventional analytical techniques.

Results: The thermal or photochemical cross-linking of these new substances resulted in hydrogel-like materials. Measurements of

cytocompatibility, hydrolytic degradability, and adhesiveness revealed a broad range of properties and low cytotoxic effects.

Discussion: Features of the cross-linked methacrylated levan derivatives are promising for the development of clinically applicable

levan based adhesives.

Conclusion: Levan derivatives are introduced as a new class of biopolymer-based macromers broadening the spectrum of available

biocompatible adhesives. Further adaption to the requirements of specific medical indications is possible especially using

biopolymer composites.

80

OP-36 PREPARATION AND PROPERTIES OF ALGINATE MUCOADHESIVE FILMS FOR ORAL

CAVITY DRUG DELIVERY

Meir Haber, Irina Lir

Biota Ltd.

Background: Oral cavity accessibility provides a rout for convenient trans-mucosal drug delivery (MDD) directly to the bloodstream

via the highly permeable oral mucosa membrane.

MDD films are vehicle that provide exact dose, large contact area, and lengthy adherence at mucosal membrane, thus enabling

enhanced drug absorption.

Aim: Evaluate the influence of composition on preparation and properties of alginate MMD films.

Materials-Methods: Five viscosity grade alginates were studied. Films were prepared by aqueous solvent- evaporation technique.

Viscosity of alginate solutions was measured using Brookfield viscometer.

Films' swelling and dissolution, and effects of plasticizers and vitamin B on films properties and release kinetics were evaluated in

saline solution at 370C.

Mechanical properties were evaluated using INSTRON at cross-head speed of 0.1mm/sec. Human volunteers tested in–vivo

mucoadhesion, and oral palate residence time.

Results-Discussion: Alginates 5% (w/v) solutions viscosity range was from 50cP to over 20,000cP (semi-gel). Heating was required

for hydration and casting high viscosity grades.

All films possess strong adherence to oral palate. Oral palate residence time was significantly affected by both film thickness and

alginate grade.

Addition of plasticizers increased elasticity, decreased strength and residency, but not affected swelling. The results indicate that

alginate have broad viscosity range significantly influence solution viscosity, film swelling and oral mucosa residence time, but have

small effect on film mechanical properties.

Conclusion: Properties of alginate films MDD are determined by alginate grade, film composition and thickness.

Part of this work was supported by grant G5RD-CT-2001-00542 from the European Union.

81

POSTER

PRESENTATIONS

82

Applications of Biomimetic Adhesives

83

PP01-01 LEVAN COATED NANO PARTICLES FOR TUMOR TARGETING DELIVERY

Esra Mutlu1, Fatemeh Bahadori

2, Hande Kazak Sarilmiser

1, Ebru Toksoy Oner

1, Mehmet S. Eroglu

3

1Department of Bioengineering, Marmara University, Istanbul, Turkey 2Department of Pharmaceutical Biotechnology, Collage of Pharmacy, Bezmi Alem University, Istanbul, Turkey 3Department of Chemical Engineering, Marmara University, Istanbul, Turkey 4TUBITAK-UME, Chemistry Group Laboratories, Kocaeli, Turkey

Background / Aim: Microbial levan is a high molecular weight, water soluble, biodegradable, anti-inflammatory and blood plasma

extender polymer1. In addition to these remarkable properties, high cell adhesion property confers its promising use in drug

delivery applications2. Cancer cells and their microenvironments need much more sugar-induced energy compared to normal cells

in order to rearrange energy metabolism, proliferation and change their shape to increase extracellular adhesion ability. Due to its

polyfructan structure, levan is expected to be more preferred energy source by cancer cells compare with normal cells. In this

study, considering its such property, levan was used as a sensor in active nano-drug carriers for cancer theraphy.

Materials-Methods: Low molecular weight levan obtained from Halomonas AAD6 (6x106 g/mol), which was used as construction

material for nano particle shells, was prepared by acid hydrolysis (16.000 g/mol) and then functionalized. Paclitaxel encapsulated

core-nano particles were prepared, using functionalized lecithin and methacrylated poly(caprolacton) (PCL-MAC) as spacer groups,

and stabilized by UV radiation. Fluorescein sodium salt was introduced between shell and core structures of the nanoparticles to

monitor them in cell culture media.

Results (quantitative and / or statistical data): Fluorescent images showed a stable core fully covered by functionalized levan

(Figure 1). Size distribution of nano particles was between 50-350 nm. To obtain uniform distribution, nano-particles were

fractionated using ultracentrifugation, and their characteristic properties such as drug loading capacity, release kinetics and cellular

uptake behaviors were determined.

Fluorescent images of stable core fully covered by functionalized levan

84

PP01-02 MUSSEL-INSPIRED DYNAMIC CROSS-LINKING OF SELF-HEALING PEPTIDE

NANOFIBER NETWORK

Hakan Ceylan, Mustafa Urel, Ayse B Tekinay, Aykutlu Dana, Mustafa O Guler

Bilkent University, National Nanotechnology Research Center

Supramolecular hydrogels benefit from stimuli-responsive, reversible gelation and self-healing following a high shear load. On the

other hand, a general drawback of such non-covalent assemblies is their weak mechanical properties. In order to overcome an

analogous challenge, mussels have adapted to a pH-dependent iron crosslinking strategy, which provides the mussel adhesive with

exceptional stiffening and self-healing properties. The present study is inspired by the mussel curing strategy to establish multiple

iron coordination points inside the self-assembled peptide networks. We synthesize catechol-functionalized cationic peptide

amphiphiles, which can assemble into nanofibers at alkaline pH. The impact of peptide-iron complexation on the morphology and

secondary structure of the supramolecular nanofibers is characterized by scanning electron microscopy, circular dichroism and

Fourier transform infrared spectroscopy. Mechanical properties of the cross-linked networks are probed by small angle oscillatory

rheology. It is shown that iron complexation is orthogonal to the self-assembly and β-sheet-driven elongation of the nanofibers. On

the other hand, iron-coordinated organic-inorganic hybrid network demonstrates higher mechanical properties comparable to that

of covalent crosslinking. Strikingly, iron cross-linking does not inhibit intrinsic reversibility of supramolecular peptide polymers into

disassembled building blocks and the self-healing ability upon high shear load. The strategy described here could be extended to

improve viscoelastic properties of a wide range of supramolecular polymer networks.

Mussel-Inspired Dynamic Crosslinking of Supramolecular Networks

85

PP01-03 BIOADHESIVE AND BIOACTIVE METHACRYLATED NANOCOMPOSITE LEVAN-BASED

BEADS FOR BONE TISSUE APPLICATIONS

Álvaro J. Leite1, Joana M. Silva

1, Özlem Ateş

2, Mehmet S. Eroğlu

3, Ebru T. Öner

2, João F. Mano

1

13B´s Research Group – Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence

on Tissue Engineering and Regenerative Medicine. AvePark, 4806-909, Caldas das Taipas, Guimarães, Portugal. ICVS/3B´s PT Government Associate

Laboratory, Braga/Guimarães, Portugal 2Marmara University, Department of Bioengineering, 34722, Istanbul, Turkey 3Marmara University, Department of Chemical Engineering, 34722, Istanbul, Turkey

Background-Aim: In bone regeneration the lack of adhesion within the defect leads to the failure of the biomaterial. A fibrous

capsule that allows micromotion to occur is formed, causing pain to patients and space for infiltration of pathogens. We propose

the development of a bioactive nanocomposite able to be implanted in bone defects with good adhesion and capable to guide

bone regeneration.

Materials-Methods: Methacrylated levan beads at a concentration of 13% (w/v) were produced by photopolymerization over

superhydrophobic surfaces. Ternary bioactive glass nanoparticles (SiO2:CaO:P2O5 (mol.%=55:40:5)) were incorporated into the

beads to promote bioactivity. Cell cytotoxicity tests were conducted using a SaOs-2 cell line. The adhesive properties of the beads

were measured and compared to control spheres of alginate.

Results: Gelation of methacrylated levan was confirmed upon 15 min. SEM images showed the presence of apatite in the

composite beads after in vitro bioactivity studies. Live/dead studies showed that after 24hours SaOs-2 cells were successfully

encapsulated and remained alive. The lap shear strength for detachment of the levan beads between the glass slides was 1.09±0.21

kPa, while the same procedure for alginate was null.

Discussion: The incorporation of bioactive glass nanoparticles in methacrylated levan hydrogels allowed materials with both

bioadhesiveness and bioactivity, and also enabled cell encapsulation. Such hydrogels may have broad applications in minimally

invasive bone tissue engineering, avoiding current approaches dependent on the use of membranes and adhesives.

Conclusion: Bioadhesive and bioactive microparticles were successfully produced and show preliminary potential to be used in

bone tissue regeneration.

86

PP01-04 ENZYMATICALLY CROSSLINKED ALBUMIN BASED BIOADHESIVE

Guy Tomer, Nadav Bramson, Orahn Preiss Bloom

LifeBond Ltd.,Caesarea, Israel

Background: Microbial transglutaminase (mTG) protein crosslinking enzyme has an extensive safety record. The benefits of mTG for

bio-adhesive crosslinking are significant in that it is non-toxic and non-inflammatory, unlike previous crosslinkers that have been

used in albumin-based bioadhesives. Albumin is a polymer of choice for use in bioadhesives due to its long record of safety in tissue

adhesives. Albumin crosslinking by mTG has been impossible due to the albumin's globular structure. The purpose of this work was

to develop a biocompatible and mechanically robust bioadhesive based on mTG-crosslinking of albumin.

Materials-Methods: Bovine serum albumin was denatured by a combination of urea, reducing agent, and heat followed by addition

of mTG. To evaluate strength of adhesion, albumin was allowed to gel between two collagen membranes and tested in a lap shear

testing system. The mechanical properties of the bioadhesive were measured in tensile testing.

Results: mTG-catalyzed gelation of albumin was demonstrated in less than 10 minutes at 37°C using unique combinations of

denaturing treatments. Maximal adhesiveness strength was 10N, maximal strain was 92%, and modulus was 31.25 kPa.

Conclusion: Globular proteins are weak substrates for mTG. Denaturing is one method where amino acid side chains become

accessible to the enzyme. However, denaturation of albumin often results in an unstable solution that precipitates. We identified

several conditions in which denatured albumin remains stable in liquid state, a prerequisite for functioning as an effective mTG

substrate. The albumin-based bioadhesive's gelation rate, mechanical, and adhesiveness properties are suited for use in a clinical

setting.

87

PP01-05 IMPROVED UNDERWATER ADHESION OF MUSHROOM-SHAPED MICROSTRUCTURE

BY ITS AIR ENTRAPMENT ABILITY

Emre Kizilkan, Lars Heepe, Stanislav N. Gorb

Functional Morphology and Biomechanics, Zoological Institute, Kiel University, Am Botanischen, Garten 9, 24118, Kiel, Germany

Background-Aim: Mushroom-shaped adhesive microstructure which is actually inspired from a terrestrial leaf beetle has an about

25% higher pull-off forces under water compared to in air [1]. It was a surprising result because van der Waals forces are

hypothesized to be lower in water [1]. Recently, it was shown that a terrestrial leaf beetle Gastrophysa viridula is also able to attach

to surfaces submerged in water by applying air bubble trapped between the setae of its adhesive pad [2]. In the present work, we

compared pull-off forces of MSAMS, measured under water with a layer of entrapped air with those completely wetted by water.

Materials-Methods: The MSAMS is made of polyvinylsiloxane (PVS). The sample surface was covered by hexagonally distributed

individual MSAMSs of about 100 μm height terminated by a thin contact plate covering about 50% of the apparent contact area [3].

Pull-off forces were measured using a custom-made setup, similar to that reported in Ref. [4].

Results: Figure 1 shows the contact interface of MSAMS in contact with a smooth cover slip at dry and different wetting states.

In Figure 2, the measured pull-off forces on the MSAMS under water with air entrapment were on average about 5.5 times higher if

compared to those measured in the fully wetted state of the MSAMS. Furthermore, the air entrapped state of the MSAMS

exhibited about 2.5 times higher pull-off forces, if compared to the dry state.

Discussion: Based on the results obtained we assume that enhanced underwater adhesion may be explained by

MSAMSs in the region of entrapped air are virtually in dry condition due to de-wetting of the contact interface.

During pull-off, the volume of entrapped air increases due to elastic deformations of individual MSAMSs. Thus, the pressure

decrease in the entrapped air spaces lead to a global suction effect [5].

Conclusion: This work has shown that under water the MSAMS is able to entrap air inducing stronger underwater adhesion than in

both dry and fully wetted conditions.

This work was funded by the German Research Foundation (DFG) under the grant scheme SFB 677-C10.

[1] M. Varenberg, and S. N. Gorb, J. R. Soc.Interface 5, 383, 2008

[2] N. Hosoda and S.N. Gorb, Proc. R. Soc. B. 279(1745), 4236-4242, 2012

[3]S.N. Gorb, M. Varenberg, A. Peressadko, and J. Tuma, J. R. Soc. Interface 4, 271-275, 2007.

[4] M. Varenberg, and S. N. Gorb, J. R. Soc. Interface, 5, 785-789, 2008

[5] E. Kizilkan, L. Heepe, and S. N. Gorb, Biological and Biomimetic Adhesives; Challenges and Opportunities.pp 65-71. RSC Publishing,

Cambridge,UK, 2013

88

Figure 1

Contact between terminal lips of MSAMS and glass cover slip (a) in dry condition, (b) underwater air entrapped and (c) underwater

full-wetted state.

Figure2

Pull-off forces of the MSAMS measured at the different wetting states at different preloads.

89

Bioadhesion: Prevention

90

PP02-01 ANTI-ADHESIVE AND ANTIBIOFILM ACTIVITY OF CRANBERRY EXTRACT

ON CANDIDA SP.

Kübra Erkan1, Demet Erdönmez

1, Necdet Sağlam

2

1Hacettepe University Institute of Gradute Studies in Science Department of Biology/Biotechnology, Ankara, Turkey

2Hacettepe University Institute of Science, Division of Nanotechnology & Nanomedicine, Ankara, Turkey

Background / Aim: Candida is commensal yeast normally present in skin, oral or vaginal flora.Cranberry juice have many various

proporties. One of these proporties,cranberry comsuption show antibacterial effect against the urinary system pathogen.Our

resarch aimed that antibiofilm anda anti-adhesive activity of cranberry extract was investigated on some clinical isolate Candidasp.

Materials-Methods: Antibiofilm activity:Candida sp. isolates were obtained hospital.Candida sp. were inoculated Sabouraud

dextrose broth(SDB)and incubated at 370C for 24 h.Then overnight cultures were diluted with SDB medium to optical density of

OD600nm Mc Farland 0,5.Antibiofilm activity assays was performed 24 well polystyrene plate. Each well was added Candida sp.

culture, SDB medium and cranberry extract.These plates incubated at 370C for 48 h. After than, plates were washed three times in

sterile 1x PBS and dried at 650C. Crystal violet was used for staining.Spectrofotometric analyse was performed at 595λ nm.Anti-

adhesive activity: We tested Candida sp. adherence to polystyrene as previously described.Overnight cultures were harvested via

centrifugation, washed twice and resuspended in PBS.Cells were inoculated in PBS containing cranberry extraction.Plate were

incubated at 370C for 2 h.Plate were washed twice with PBS.XTT assay was used to measure the anti-adhesive activity. Results

(quantitative and / or statistical data):C.albicans biofilm inhibition percent was determinated as a %90,Anti-adhesive activity

measured OD490nm.

Discussion: Other Candida sp. was showedvarious biofilm inhibition and anti-adhesive activity ratio.It depend on molecular

proporties of speciesConclusion: We suggest that cranberry in Candida species inhibition of biofilm formation and anti-adhesive

activity.Cranberry can be used as drug supplement for strengthening of the human immune system.

91

PP02-02 INVESTIGATION OF ANTIBIOFILM ACTIVITY USING DIFFERENT SPIRULINA SP.

EXTRACTS

Minoo Pourhassan Shamchi1, Demet Erdönmez

1, Kübra Erkan

1, Necdet Sağlam

2, Hikmet Katırcıoğlu

3

1Hacettepe University Institute of Science Department of Biology/Biotechnology, Ankara, Turkey 2Hacettepe University Institute of Science, Division of Nanotechnology & Nanomedicine, Ankara, Turkey 3Gazi University, Faculty of Education, Department of Biology Education, 06500, Teknikokullar, Ankara, Turkey

Background-Aim: Microbial biofilms are responsible for several chronic diseases that are difficult to treat as a cystic fibrosis,

endocarditis, and cystitis. Microorganisms which occurs biofilm show much greater resistance to antibiotics than their free-living

counterparts. In this study antibiofilm activity of different Spirulina sp. extractions were investigated against Candida

albicans 10231.

Materials-Methods: C. albicans was inoculated in Sabouraud Dextrose Broth (SDB) and incubated at 37o C for 24 hours. Overnight

cultures were diluted by SDB to optical density of OD 600 nm Mc Farland 0.5. DifferentSpirulina sp. extracts were prepared.

Methanol, Dimethyl Sulfoxide (DMSO), Acetone and Chloroform used as solvants. Polystyrene 24- well plates were used for

antibiofilm activity assays. Each well filled with 800 µl SDB, 100 µl C. albicans 10231 and 100 µl of Spirulina sp. extractions.Plates

incubated at 37oC for 48 hours plates then discarded and nonadherant cells washed away using sterile 1×PBS three times and dried

at 65oC. Crystal Violet was used for staining. Samples were measured at 595ƛ nm by spectrophotometer.

Results (quantitative and / or statistical data): According to results Spirulina sp. extracts showed antibiofilm activity against C.

albicans 10231. We observed maximum inhibition percent of 90% in chloroform extract. This inhibition ratio followed by Methanol,

DMSO and acetone extracts 81%, 73% and 65% respectively.

Conclusion: This study confirmed that different Spirulina sp. extracts with antimicrobial and antioxidant activity could be used as a

promising antibiofilm agent.

92

Bioinspired Adhesive Designs

and Formulations

93

PP03-01 BIO-INSPIRED ORIENTATION DEPENDENT FRICTION

Longjian Xue1, Jagoba Iturri

1, W. Jon. P. Barnes

2, Michael Kappl


1, Aránzazu Del Campo

1

1Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128-Mainz, Germany 2Centre for Cell Engineering, University of Glasgow, Glasgow G12 8QQ, UK

Spatula-shaped contact elements are involved in most biological hairy adhesive systems, like in gecko’s attachment pads. The

anisotropic geometry is expected to provide directionality in the adhesive and frictional properties and facilitates switching

between strong attachment and effortless detachment during locomotion. Inspired by gecko’s toe pad design, we study friction of

polydimethylsiloxane (PDMS) micropillars terminated with symmetric (T-shaped) and asymmetric (spatula shaped) overhangs.

Friction forces upon shearing towards and against the spatular end were evaluated and compared with friction forces on T-shaped

pillars and pillars without overhangs. The shape of the friction curves and the values of the friction forces on spatula-terminated

pillars were orientation-dependent. Kinetic friction forces were enhanced when shearing against the spatular end, while static

friction was stronger in the direction towards spatular end. The overall friction force was higher in the direction against the spatula

end. The maximum value was limited by the mechanical stability of the overhangs during shear. The aspect ratio of the pillar had a

strong influence on the magnitude of the friction force and its contribution surpassed and masked that of the spatular tip for aspect

ratios >2.

94

PP03-02 BIO-INSPIRED ADHESIVE DESIGN FOR FLOODED CONDITIONS

Jagoba Iturri1, Longjian Xue

1, Michael Kappl

1, W. Jon. P. Barnes


1, Aránzazu Del Campo

1

1Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128-Mainz, Germany 2Centre for Cell Engineering, University of Glasgow, Glasgow G12 8QQ, UK

Tree- and torrent-frogs are able to adhere and walk onto wet and flooded surfaces respectively. The complex hierarchical

superstructure of their attachment pads, containing soft micro and nanopillars separated by narrow channels of defined

dimensions, is the key for their skills. Significant differences in the adhesive designs of tree and torrent frogs are found in nature,

which most probably correlate with differences in their adhesive performance, in particular in the presence of thick films of water.

We have prepared elastomer-based artificial mimics of tree- and torrent-frog attachment microstructures (regular and elongated

hexagonal patterns, respectively, separated by 1 m wide channels). We have studied the friction performance of the fabricated frog

mimics under different scenarios: dry, wet and flooded environments, varying surface energies, increasing preload forces or liquid

volumes, among others. Results show a strong dependence of friction performance on the pattern design. Torrent-frog toe pad

mimics show an enhanced performance in all the cases, independently to the presence of a thin film of water at the interface.

Elongated patterns present directional friction, which might be important implications for the detachment process.

95

PP03-03 CHOLESTEROL AS SYNTHETIC BUILDING BLOCKS FOR ARTIFICIAL GLYCOLILIPIDS

WITH CHARACTERISTIC CHEMICAL PROPERTIES

Fahima Ali Rachedi1, Fahima Ali Rachedi

2

1Département des Sciences, Centre Universitaire, 41000, Souk-Ahras, Algérie 2Laboratoire de Chimie Organique Appliquée, Université Badji-Mokhtar, BP 12, 23000, Annaba, Algérie

Cholesterol is one of the most widely distributed natural materials and has a unique chemical structure such as a steroid skeleton.

Many types of chemical transformations of cholesterol functional groups have been developed. There is an interest in the

derivatization of cholesterol and to introduce alkyl branched fatty acids into the molecule. The C3-hydroxy group allows the

preparation of various cholesteryl compounds such as cholesteryl esters or ethers. Such compounds have found applications in the

formulation of cosmetics and toiletries over the past few decades. An extraordinary interesting case is related to cholesteryl esters

and their use in gene therapy delivery systems. These results can be attributed to their potential for forming cell-mimic

membranes, because cholesterol is the most important building block of living cell membranes. In terms of organic synthesis,

cholesterol is a strategically useful material. A typical case is remote functionalization by chemical reactions or by biocatalysis. In

the future, cholesterol should be considered as a key compound, a building block for the construction of artificial lipid-like

membranes by self-assembly. Also, as cholesterol is one of the members of the fat and oil family, fat and oil chemists should study

and develop cholesterol chemistry even further.

The work presented here will focus on the synthesis of a new series of cholesteryl ethers containing alkyl branched acyl groups.

96

PP03-04 E-TUNED NANO- AND MICROHAIRS FOR BIO-INSPIRED ADHESIVES

Julia Syurik, Johanna Wolf, Budr Albakri, Marc Schneider, Hendrik Hölscher

Institute of Microstructure Technology, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany

The discovery of Gecko’s adhesion mechanism inspired an amount of works on biomimetic adhesives, where principles of

geometrical hierarchy and contact splitting are used. However, the understanding of hierarchy might be extended from geometrical

to functional hierarchy where a gradient of desired properties is presented. It was recently discovered that the legs of some insects

have microhairs that are strong, rigid and flexible at the same time due to a gradient (hierarchy) of their elastic modulus up to

several orders of magnitude. Such structures are extremely interesting prototypes to fabricate bio-inspired adhesives.

We offer an approach, where a gradient of elastic modulus is achieved by using porous polymers of controlled pores size. Porous

polymers belong to a large and important category of industrial plastics. Porosity can be introduced into a wide variety of polymers

(cross-linked elastomers, PS, PMMA etc.) during polymerization or post processing treatment in presence of technological gases.

Mechanical properties of porous polymers, including the elastic modulus, depend on density and size of the pores. The latter can

be controlled via processing parameters, and a gradient of processing parameters allows fabrication of the microhairs with a

tunable elastic modulus.

First, PMMA porous foil was prepared by saturation of a flat PMMA foil in CO2 with further temperature treatment. Second, insect-

inspired PMMA adhesives with porous microhairs were prepared via hot-embossing. The morphology and adhesion of the insect-

inspired PMMA adhesives was investigated.

97

PP03-05 BIOMIMETIC PHOSPHORYLATED CO-POLYPEPTIDES WITH CONTROLLED DEGREE OF

PHOSPHORYLATION

Vitali Lipik, Lihong Zhang, Ali Miserez

School of Materials Science and Engineering, Nanyang Technological University

Phosphorylated polypeptides represent promising biomimetic adhesives. However to date, large-scale synthesis of phosphorylated

polypeptides with controlled degrees of phosphorylation has not been achieved. Here, a co-polypeptide synthesis strategy based

on the Ring-Opening Polymerization (ROP) of N-carboxyanhydrides (NCAs), followed by controlled phosphorylation of serine (Ser)

residues, will be presented. The molecular design, including amino acid composition and molecular weight of polypeptides,

mimicked the intriguing phosphorylated protein Pc-3 secreted by the sandcastle tube worm Phragmatopoma californica. Three NCA

monomers were synthesized, namely Ser with free –OH groups, and Ser and Tyr with protected –OH groups, and subsequently

polymerized with various feeding ratios in order to obtain a broad range of final amino acid compositions. In the final step,

phosphorylation targeting free –OH groups of Ser was conducted. With this strategy, the degree of phosphorylation is governed by

the initial amount of unprotected –OH groups of the precursor Ser–NCA, and the final co-polypeptides contain relative amounts of

Tyr and pSer that can be tailored, yielding a composition and molecular weight (MW) that closely match those of Pc-3. This control

of phosphorylation leads to polypeptides exhibiting a wide range of zeta potential values between -20 and -50 mV. Using Dynamic

Light Scattering (DLS), Surface Plasmon Resonance (SPR), and Quartz Crystal Microbalance with Dissipation (QCM-D), we

demonstrate that these phosphorylated polypeptides exhibit affinity towards divalent ions such as Ca2+, thus opening the door for

their usage as scaffolds for mineralized tissue repair or as a major component of biocompatible adhesives.

This research is funded by the Singapore National Research Foundation (NRF).

98

PP03-06 THE OPTIMIZATION OF CULTURE CONDITION FOR THE HIGH EFFICIENCY

PRODUCTION OF A SELF-SILICIFIYING FUSION PROTEIN, GFP-R5, IN E. COLI

In Wong Song1, Byung Chun Kim

2, Hyojung Park

2, Byoung In Sang

1

1Department of Fuel cell and Hydrogen Technology, Hanyang University, 17 Hangdang-dong, Seongdong-Gu, Seoul 133-791, Republic of Korea 2Department of Chemical Engineering, Hanyang University, 17 Hangdang-dong, Seongdong-Gu, Seoul 133-791, Republic of Korea

Diatoms, which are aquatic, photosynthetic, unicellular algae, have gained much attention owing to their intricately and

hierarchically nanostructured silica cell walls. The silaffin polypeptides derived from Cylindrotheca fusiformis are major

components of the intricate silica cell wall. They are heavily post-translationally modified proteins responsible for silica deposition

at ambient temperature and pressure. Recently, it was discovered that an unmodified silaffin peptide, the R5 peptide

(H2NSSKKSGSYSGSKGSKRRIL-COOH) identified as a repeat unit of the silaffin polypeptide showed the same silica precipitation

activity when added to a silicic acid solution under ambient conditions. In previous research, the R5 peptide was successfully

applied to immobilize biomolecules in silica materials. In addition, they were fused with proteins for novel nanocomposite

materials and for immobilization. Recently, we constructed a fusion protein consisting of R5 peptide and glucose oxidase for

biosensors. Such a “smart glue” can potentially seal surface defects and dentinal tubules to reduce risk of tooth decay and dental

hypersensitivity. It would be interesting to explore whether limited expression of silicatein on these cells and subsequent

depositioning of biosilica directly on the cell surface could stimulate osteoblast mineralization even further, though there is an

obvious disadvantage in the continuous expression of silicatein and potential for unwanted biosilica formation

unless the precursor supply is removed. Like this, the silicification has been utilized in diverse industrial applications. It can also be

applied for the production of catalyst template, dental adhesives, electrode for battery or electrochemical systems. One of

silicification methods is using the polycationic R5 peptide produced by the diatom Cylindrotheca fusiformis. To extend the

biosilicification to various applications, the key protein of biosilicification, R5, should be produced with easy process. Therefore, in

this study, a GFP-R5 fusion protein containing R5 peptide can be transformed to self-silicified nano particles. To be focused on

enhancive production of GFP-R5 fused protein, it is useful way to increasing the efficiency of silicification. Concentration of IPTG

and temperature were important factors to be considered in order to produce GFP-R5 fused protein. In this study focused that two

factors affected on the production of GFP-R5 fused protein. Result of the experiment, the most production of GFP-R5 fused protein

was verified at 25 ℃, with 0.5 mM IPTG.

99

Catechol-Based Adhesives

100

PP04-01 REVERTING DOPA ADHESIVENESS

Julieta Paez, Oya Ustahüseyin, Aránzazu Del Campo

Max-Planck-Institut für Polymerforschung. Ackermannweg 10, 55128, Mainz, Germany

Mussel-inspired, DOPA functionalized biomaterials have the ability of crosslinking and adhering to different substrates under

physiological conditions. These have been applied over the last decade as biocompatible coatings, cell matrices or surgery glues. In

nature, variants of DOPA containing different substituents in the catechol unit are present. These small chemical variations confer

additional properties to catechol derived materials. For example, NO2-substitution of the catechol ring allows light-triggered

degradation of nitrocatechol crosslinked networks, which can be used to reverse their adhesiveness. Up to now the light doses

required for photodegradation are too high to be compatible with biological material.(1) We present new nitro-catechol derivatives

with higher photosensitivity that are compatible with embedded cells.

(1) Z. Shafiq, J. Cui, L. Pastor-Pérez, V. San Miguel, R. A. Gropeanu, C. Serrano, A. del Campo. Angewandte Chemie International Edition, 51, 4332–

4335 (2012).

101

Chemical and Structural Characterization

of Natural Adhesives

102

PP05-01 BIOADHESION IN ASCIDIANS

Ute Rothbächer

Department of Evolution and Developmental Biology, Zoological Institute, University Innsbruck, Austria

Background-Aim: Adhesive organs in ascidians are important for attachment of the swimming larva, trigger metamorphosis to

sessile juveniles and settlement of adults. The developmental origin of adhesive papillae is layed down very early in development

within the neuroectodermal domain of the 32 cell stage. An invariable cell lineage and advanced molecular tools including

sequenced genomes for both Ciona intestinalis and recently Phallusia mammillata, make it possible to analyse adhesive organ

formation and glue production from its beginnings, with cellular resolution and in a comparative fashion.

Materials-Methods: By gain-of-function (electroporation in ascidian zygotes) and loss-of function (Morpholino injection) we are

presently analyzing the influence of MAPK/Ets factor signalling at the early inductive phase and fate progression of palp precursors.

As these factors also play a role in metamorphosis triggered by adhesive papillae at the time of larva settlement, we apply these

functional approaches and tools as entry point to test for a number of genes identified in differential screens with adhesive organs

(effected or not in papillae-cut larvae, cell-division requiring events for tunic expansion and adhesion).

Results and Discussion: The ascidian model organism and its advantages will be presented, as well as an overview on what is

known in molecular terms about palp formation in ascidians. Furthermore, our data on initial palp precursor formation and the

involvement of an Ets factor network herein will be shown. The novel approaches to analyse gene networks and final products in

adhesive organs (palps, epidermis) will be presented and discussed.

103

PP05-02 ULTRASTRUCTURE OF TARSAL “JUMPING PADS” IN LEAFHOPPERS

Hanns Hagen Goetzke1, Oliver Betz

2, Walter Federle

1

1Department of Zoology, University of Cambridge, Cambridge, United Kingdom 2Institut für Evolution und Ökologie, Eberhard Karls Universität Tübingen, Tübingen, Germany

Background: Leafhoppers (Hemiptera: Cicadellidae) possess special soft attachment structures (platellae) on their hind legs' first

two tarsomeres which allow them to jump from smooth surfaces. Platellae produce large friction forces when pushed rapidly. This

study investigated the platellae's ultrastructure.

Materials- Methods: Platellae on Aphrodes sp. leafhoppers' hind legs were freeze-fractured and images recorded with a scanning

electron microscope (SEM). Hind legs were dehydrated in ethanol and critical point dried before SEM. Moreover, thin sections of

embedded platellae were prepared for light and transmission electron microscopy.

Results: Aphrodes sp. leafhoppers possess a row of 5-6 platellae on the first tarsomere's distal end and a row of 2-3 platellae on the

distal end of the second. Platellae protrude from the base of short and thick spines; platellae are 50-85 μm long and tubular with a

thick cuticular wall. On the ventral side, they showed a thick amorphous epicuticle and a procuticle with fine cuticular rods forming

a sponge-like structure, whereas on the dorsal side, the epicuticle was thinner and the procuticle was more sclerotized.

Discussion: The cuticle on the platellae's ventral side with its sponge-like ultrastructure is similar to cuticle of smooth pads of

insects, which can deform around asperities on rough substrates and generate large friction and adhesion forces on diverse

surfaces.

Conclusion: Leafhoppers' platellae are soft attachment structures allowing them to jump from smooth surfaces. Their sponge-like

cuticular ultrastructure is similar to that of smooth adhesive pads, suggesting that both pad types have been shaped by similar

selection pressures.

This research project was partially supported by a Short Term Scientific Mission to Hanns Hagen Goetzke (ECOST-STSM-TD0906-

250813-034958).

104

PP05-03 LOCALIZATION OF THE CELLULAR ORIGIN OF SEA STAR ADHESIVE PROTEINS

USING IN SITU HYBRIDIZATION

Elise Hennebert1, Birgit Lengerer

2, Mélanie Demeuldre

1, Peter Ladurner

2, Patrick Flammang

1

1Laboratory of Biology of Marine Organisms and Biomimetics, Research Institute for Biosciences, University of Mons, 7000 Mons, Belgium 2Institute of Zoology, University of Innsbruck, 6020 Innsbruck, Austria

Background: Sea stars adhere to various surfaces using an adhesive material secreted by their tube feet. Recently, proteins

extracted from this material were analyzed using mass spectrometry and the resulting data were compared to the translated tube

foot transcriptome. For each protein identified in the transcriptome, the emPAI, an index which estimates the relative abundance

of proteins in a mixture, was calculated. This allowed to identify the most abundant proteins. The aim of my stay in the Institute of

Zoology (University of Innsbruck, Austria) was to use the technique of in situ hybridization to confirm the role of some of these

proteins in sea star adhesion through their localization in the cells producing the adhesive material, the so-called adhesive cells.

Methods: The complementary DNA sequences coding for these proteins picked up in the tube foot transcriptome were used to

design specific oligonucleotide probes. These probes were used in whole tube feet in situ hybridization to label the corresponding

mRNA in the cells.

Results: For the 4 proteins investigated, the results show a specific labelling at the base of the tube foot disc, at the level of the

adhesive cell nuclei.

Discussion: These results clearly demonstrate that the proteins identified as major components in the adhesive material using mass

spectrometry are synthesized in the adhesive cells and therefore probably correspond to adhesive proteins.

Conclusion: In a near future, the technique of in situ hybridization will be developed in my host laboratory and will be applied to

the other major adhesive proteins.

105

PP05-04 IDENTIFICATION OF ADHESION CANDIDATE GENES IN FLATWORMS

Julia Wunderer, Birgit Lengerer, Robert Pjeta, Marcelo Rodrigues, Willi Salvenmoser, Peter Ladurner

Institute of Zoology and Center of Molecular Bioscience Innsbruck, University of Innsbruck, Technikerstr. 25, A-6020 Innsbruck, Austria

Background-Aim: Free living marine flatworms are able to attach temporarily to the substrate by means of specialized adhesive

organs. Macrostomum lignano is an emerging model organism for bioadhesion because of the availability of molecular tools, easy

culturing and knowledge of the morphology of the adhesive system. M. lignano possesses about 130 adhesive organs in the tail

plate allowing the animals to repeatedly adhere and release. Each adhesive organ comprises three different cell types: one

adhesive gland, one releasing gland and one modified epidermal cell, the anchor cell. The aim of the study is to identify genes and

further the proteins that are involved in the adhesive and releasing process.

Materials-Methods: Genes mainly expressed in the tail plate of M. lignano were screened by whole mount in situ hybridization.

Results and Discussion: In total 239 tail specific genes were screened. 29 genes (12%) showed an expression in the region of the

adhesive organs. Other tail-specific expression could be found in organs belonging to the reproductive system, namely the female

opening, the prostate gland and the stylet. Non tail-specific expression was obtained in the region of the gonads and the rhabdite

glands.

Gene knockdown by RNAi combined with bioinformatical analysis should enable to investigate the function of the candidate genes.

Conclusion: Molecular tools comprise a variety of methods to reveal molecules involved in temporary adhesion in M. lignano and

other organisms.

Supported by FWF grant P25404-B25 and COST Action TD0906

106

PP05-05 THE ELABORATE ADHESIVE SYSTEM OF PROSERIATE FLATWORMS

Robert Pjeta, Julia Wunderer, Birgit Lengerer, Marcelo Rodrigues, Willi Salvenmoser, Peter Ladurner

Institute of Zoology, CMBI, University of Innsbruck, Austria

Background-Aim: Many animals of the interstitial fauna are able to temporarily adhere by the use of specialized structures. Among

flatworms, Proseriates have developed a highly effective adhesive ability that allows them to live in high energy environments such

as exposed sandy beaches. In our investigations we aim to morphologically characterize the adhesive system of two proseriate

flatworm species, the Mediterranean Pseudomonocelis sp. and the Red Sea flatworm Minona ileanae.

Materials-Methods: The morphology of the adhesive system was examined by scanning (SEM)- and transmission (TEM) electron

microscopy using standard preparation methods. Samples were observed with a Zeiss Libra 120 (TEM) and a Zeiss DSM 950 (SEM).

Results and Discussion: Proseriate adhesion relies on a duo-gland system including adhesive gland cells, releasing gland cells and

modified epithelial cells. The necks of adhesive glands branch several times and build up a meshwork. Together with releasing

gland necks they protrude out of the epidermis via several individual epithelial cells and form cushion-shaped papillae.

Conclusion: Our work showed that proseriate adhesion is relying on a complex meshwork composed of adhesive gland cells. This,

in combination with cushion-shaped adhesive papillae possibly reflects on the higher adhesive ability of proseriates compared to

other flatworms. Further investigations regarding the molecular composition of the adhesive material are planned.

Supported by FWF grant P25404-B25, COST Action TD0906 and a PhD fellowship of the University of Innsbruck

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PP05-06 DIRECT SOLUBILIZATION OF PHOSPHORYLATED PROTEINS FROM LARVAL

ADHESIVE NET OF CADDISFLY, STENOPSYCHE MARMORATA

Kousaku Ohkawa1, Takaomi Nomura

2, Ryoichi Arai

2, Kimio Hirabayashi

2, Masuhiro Tsukada

2, Koji Abe

3

1Institute of High Polymer Research, Faculty of Textile Science and Technology, Shinshu University 2Division of Applied Biology, Faculty of Textile Science and Technology, Shinshu University 3Division of Chemistry and Materials, Faculty of Textile Science and Technology, Shinshu University

Background-Aim: We recently reported the purification and characterization of underwater silk/cement proteins from

caddisfly, Stenopsyche marmorata (Figure 1). The S. marmorata silk proteins (Smsps)-1 has a long-range periodic sequence,

including highly phosphorylated sites. The major purpose of this study is to solubilize silk/cement proteins directly from larval net

and to discuss the localization of Smsps-1-4.

Materials-Methods: Larval nets were minced in DW. Proteins were extracted with a Tris buffer containing 10 mM DTT and 1% SDS

(Ext1). The remaining proteins were solubilized with the same buffer containing 5 mM EDTA (Ext2). As-spun net and Ext2 residue

(Pt2) were subjected to SEM-EDX (Figure 2).

Results: Smsp-1&4 were found in Ex2 with no other proteins, indicating that Ca2+ mediates association of Smsp-1&4. The molar

percentages were (P, S, Ca) = (0.87%, 0.44%, 0.69%) and (5.00%, 3.45%, 0%) for as-spun net and Pt2, respectively.

Discussion: The presence of calcium in the as-spun net and the co-solubilization of Smsp-1&4 by EDTA indicate that Ca2+

mediates

association of two proteins. The immuno-staining using anti-pSer-antibody suggested that Smp-1&4 are only two phospho-

proteins, which work as cements and form silk fibers. The broken tube-like shape of Pt2 implies that the fiber-inner matrices are

composed of Smsp-1&4 and coated with Pt2.

Conclusion: The localization of Smsp-1&4 was indicated in this study. Previously, we reported the absence of DOPA in the silk gland

proteins of the caddisfly. Tricopetrans, hence, adopt the strategy different from mussels or polychaeta, which combines DOPA

and pSer.

This work was supported by Grants-in-Aid No. 22350103, No. 23651083 for KO, No. 22510028 for KH and No. 22580060 for MT from

MEXT and part was supported by the Japanese Association for Marine Biology (JAMBIO) No 23–73.

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Figure 1

Laboratory culture of caddisfly, S. marmorata.

Figure 2

SEM images of as-spun net and Pt2.

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PP05-07 BYSSUS PRECURSOR PROTEINS FROM ASIAN FRESHWATER MUSSEL, LIMNOPERNA

FORTUNEI

Kousaku Ohkawa1, Masakazu Hachisu

2, Takaomi Nomura

3

1Institute of High Polymer Research, Faculty of Textile Science and Technology, Shinshu University 2National Institute of Advanced Industrial Science and Technology, Hokkaido Center 3Division of Applied Biology, Faculty of Textile Science and Technology, Shinshu University

Background/Aim: Asian freshwater mussel, Limnoperna fortunei (Figure 1) adopts the adhesive organ similar to mussels. A byssus

precursor protein, L. fortunei foot protein (Lffp)-1 was found in 1999. Lffp-1 preserves the DOPA-containing decapeptide motif,

while has enriched electrolytic residues. The present work is conducted for searching other fp family to discuss its adaptation to

underwater adhesion in freshwater.

Materials-Methods: A cDNA library was constructed from L. fortunei foot mRNAs, and 192 ESTs were subjected to BLAST search in

UniProt to find homologues to other mussel species.

Results: Two clones were registered in DDBJ with accession codes, AB910939 and AB910940 (Figure 2). AB910939 (635 bp) encodes

repetitive regions homologous to fp-2s from Mtylus edulis (E, 2E-38) and M. garoprovincialis (6E-38), and is tentatively named Lffp-

2. The pI of Lffp-2 is 6.92, while those of Mgfp-2 and Mefp-2 are 9.21 and 9.16, respectively. AB910940 is a putative tyrosinase.

Discussion: The ionic strength in freshwater is lower than seawater, so that electrostatic interactions between Asp/Glu and Lys/Arg

is enhanced in freshwater. This is a profitable factor for protein aggregation to achieve rapid adhesion, while stronger ionic

strength in seawater can increase the inductive dipolar interaction between the hydrophobic residues in marine mussels’ fps.

Conclusion: The key components in the mussel adhesion is DOPA, which is conserved both in marine and freshwater mussels. In

addition to this, the glue proteins have been diversified probably with adapting the appropriate amino acids to express more stable

adhesive strength on various substrates.

This work was supported by Grants-in-Aid No. 22350103, No. 23651083 for KO, No. 22510028 for KH and No. 22580060 for MT from

MEXT and part was supported by the Japanese Association for Marine Biology (JAMBIO) No 23–73.

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Figure 1

Construction of cDNA library from L. fortunei foot.

Figure 2

Nucleotide sequence of AB910939 (Lffp-2)

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PP05-08 CHARACTERIZATION OF CARBOHYDRATES IN FLATWORM ADHESIVE ORGANS

Birgit Lengerer1, Marcelo Rodrigues

1, Elise Hennebert

2, Willi Salvenmoser

1, Patrick Flammang

2, Peter Ladurner

1

1Institute of Zoology, CMBI, University of Innsbruck, Austria 2Laboratory of Biology of Marine Organisms and Biomimetics, University of Mons, Belgium

Background-Aim: The marine flatworm Macrostomum lignano exhibits approximately 130 adhesive organs, to attach and stabilize

itself. The adhesion is temporary and the worm attaches and releases several times within a minute. Each adhesive organ consists

of three different cell types, two secretory glands and one modified epidermal cell. To identify carbohydrate components, we used

15 lectins to stain whole mount animals. These proteins bind specifically to oligosaccharidic motifs found in glycoproteins.

Material-Methods: After fixation, washing and blocking, 15 biotinylated lectins were applied to whole mount specimens. Bound

lectins were visualized using Texas-Red-conjugated streptavidin. Control reactions were performed by using the lectins saturated

with their inhibitory monosaccharide.

Results: Out of 15 tested lectins, 9 labelled different cell types of M. lignano, including epidermis, pharynx and different secretory

glands. Two lectins (PNA and RCA120), both recognizing glycoconjugates containing galactose residues, labelled the secretory

glands of the adhesive organs. This staining was inhibited by a pre-incubation of the two lectins with D-galactose.

Discussion: Glycosylation of proteins can play an essential role in the adhesives of permanently, transitorily and temporarily

attaching animals. We were able to identify glycoconjugates containing galactose residues in the secretory glands of M. lignano,

probably indicating the presence of at least one glycoprotein.

Conclusion: Our results enable the further investigation of the role of glycoproteins in flatworm adhesion.

Funding: Supported by FWF grant P25404-B25 and COST Action TD0906. BL is a recipient of a DOC Fellowship of the Austrian

Academy of Sciences at the University of Innsbruck.

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PP05-09 HIGH RESOLUTION ATOMIC FORCE MICROSCOPY OF NATIVE ENTOBDELLA SOLEAE

ADHESIVE PROTEINS

Alma Salibasic, Suzi P. Jarvis

Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin, Ireland

While amyloid, highly ordered cross-β sheet protein aggregate is frequently associated with neurodegenerative diseases such as

Alzheimer’s and Parkinson’s disease, it also plays a functional role in many natural materials including adhesives, biofilms and

hydrophobic coatings. Here, we study Entobdella soleae a protein-based adhesive of a parasitic marine flatworm This tissue

adhesive has specific attachment-detachment mechanism which works instantly and reversibly providing remarkable adherence to

wet surfaces.

We use single molecular approach to study and characterize the adhesive. We collected the secreted adhesive by manually

manipulating live parasites to extrude adhesive prints from their anterior end onto a clean glass slide. Further, the frequency

modulation atomic force microscopy (FM-AFM), a highly sensitive, true atomic scale resolution method was used for molecular

level imaging of footprints in liquid and finally the single molecular force spectroscopy was used to pull the molecule apart and

study intermolecular and intramolecular forces.

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PP05-10 ANALYSIS OF SACCHARIDES, OCCURRING ON TIBIA AND TARSAL SURFACES

OF GROMPHADORHINA PORTENTOSA

Beate Stahl1, Klaus Albert

2, Christian Zwiener

1

1University of Tuebingen, Environmental Analytical Chemistry, Center for Applied Geoscience 2University of Tuebingen, Institute of Organic Chemistry

Background-Aim: The carbohydrate composition of the adhesive tarsal secretion of cockroach Gromphadorhina-portentosa was

characterized and compared to the composition of the non-adhesive cuticula on tibia. For the adhesive system a fluid-film mediates

the contact between the tarsi and the substrate. It is presumed that this fluid is an oil-in-water emulsion and that carbohydrates

are physically affecting the surfactant properties.

Materials-Methods: The samples were taken from living cockroaches (Fig. 1), prepared and analysed with HPLC-ESI-MS/MS.

Samples aliquots were hydrolyzed to determine the bound saccharides. Derivitisation with phenyl-methyl-pyrazolone was used for

the LC-ESI-MS/MS analysis.

Results: The distribution of saccharides on tibia and tarsi is similar (chain lengths from 2 to 5). Monosaccharides have the highest

concentrations. Hydrolysis increased the concentration of monosaccharides to a differing extent for the tibia (Fig. 2) and tarsi.

Discussion: For both sample-types, the amount of the saccharides decreased with the increase of molecular weight. Hydrolysis

increased the concentration of monosaccharides more than what can be explained by the decrease of the oligosaccharides.

Therefore bound saccharides are expected to occur and differences between the tibia and tarsi were observed.

Conclusions: Differences in the carbohydrates were found when comparing the secretions of the adhesive tarsi and non-adhesive

tibia of the cockroach Gromphadorhina-portentosa.

The developed methods of sampling, sample preparation, and analytical measurement support a significant contribution to the

understanding of the distribution of saccharides on tibia and tarsi.

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Gromphadorhina portentosa

Sampling with glass-wool on a tibia of a living Gromphadorhina portentosa.

Tibia

Concentration of saccharides on tibia before and after hydrolysis. The increase of the monosaccharaides 1-hexose and 1-pentose

with hydrolysis hint to bound saccharides.

Acknowledgement: Project „Functional analysis of adhesive locomotion and catching pray structures“ is funded by the DFG.

115

PP05-11 NEW EVIDENCES THAT SEA URCHIN'S ADHESIVES SHARE COMMON PROTEINS

Raquel Mesquita Ribeiro1, Cristina Ribeiro Da Silva

1, Gonçalo Conde Da Costa

1, Carlos Cordeiro

1, Romana Santos

2

1Centro de Química e Bioquímica, Departamento de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa, Lisboa, Portugal 2Unidade de Investigação em Ciências Orais e Biomédicas, Faculdade de Medicina Dentária da Universidade de Lisboa, Portugal

Background-Aim: Sea urchins possess specialized adhesive organs - adoral tube feet, for strong but reversible attachment. These

are composed of an adhesive disc and an extensible stem, which have a similar tissue stratification, except that the disc encloses

the cells that contain the adhesive and de-adhesive secretions. Previous immunohistochemical data suggested that sea urchin

adhesives don’t share common epitopes and thus would be species-specific. To verify this hypothesis,the same antiserum, raised

against the adhesive of Sphaerechinus granularis, was used to search for antibody-cross reactivity not on histological sections, but

on western-blots of Paracentrotus lividus and Arbacia lixula tube feet discs protein extracts.

Methods: 1D and 2D electrophoresis, western-blotting, mass spectrometry.

Results: We demonstrate that although raised against S.g. adhesive, the antiserum specifically labels tube feet disc extracts from

this species that contain the adhesive precursors. When used on tube feet discs extracts from P.l. and A.l, positive labeling was

obtained in both species, but P.l., exhibited a higher number of common epitopes with S.g. adhesives. Common proteins were

identified by LC-MS/MS, confirming that sea urchins adhesives do share common traits and therefore are not species-specific.

Discussion: These apparently contradictory results might be explained by the fact that although histological sections are

permeabilized to allow antiserum penetration, little or no labeling might be observed if the number of common epitopes is

reduced. However, if proteins are denatured, as in western-blots, the chances of exposing more epitopes is enhanced, clearly

demonstrating the need to use complementary techniques to study biological adhesives.

116

PP05-12 ‘UN-STICKING’ A WHITE’S TREE FROG’S TOE PADS: A MECHANISM THAT ENSURES

EASY DETACHMENT DURING WALKING AND CLIMBING

Diana Samuel, Thomas Endlein, Jon Barnes, Mathis Riehle

Centre for Cell Engineering, University of Glasgow, Scotland UK

Tree frogs can adhere to a variety of surfaces using a temporary adhesive mechanism called ‘wet adhesion’. In keeping with the

principles governing temporary adhesives, this should enable them to both ‘stick’ their adhesive toe pads adequately and detach

them effortlessly. To investigate the latter, we have developed 2 systems: one allows the visualization of toe pad contact area

alone, whilst the other combines this with the simultaneous measurement of forces exerted by the pads along all 3 axes (normal,

fore-aft and lateral). These systems can be used during walking and climbing. Data acquired using the first system revealed that

White’s tree frogs (Litoria caerulea) are able to detach their toe pads rapidly; the speed of detachment is generally conserved –

irrespective of whether the frog is walking or climbing. This ability is due to the mode of detachment: peeling. Although there are

many ways in which their toe pads can be peeled from a surface – influenced somewhat by the angle of the toe with respect to the

body – they are generally peeled longitudinally, from the rear to the front. Using the second system, we were able to establish that

only a negligible amount of force is exerted during detachment, to overcome the adhesive force. In addition, detachment is often

preceded by a forwards-acting force, which may serve to initiate peeling. Therefore, the detachment mechanism utilized by this

tree frog – rapid peeling with minimal force – ensures the quick and (almost) effortless removal of its toe pads.

117

PP05-13 STATIC AND DYNAMICS PROPERTIES OF LEVAN AS A FUNCTION OF SALT

CONCENTRATION VIA MOLECULAR DYNAMICS SIMULATIONS

Binnaz Coskunkan1, Gulcin Cem

2, Seyda Bucak

3, Deniz Turgut

2, Deniz Rende

2, Nihat Baysal

2, Rahmi Ozisik

2, K. Yalcin Arga

3,

Ebru Toksoy Oner3

1Department of Chemical Engineering, Yeditepe University, Istanbul 34755, Turkey 2Department of Materials Science and Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA 3Department of Bioengineering, Marmara University, Istanbul 34722, Turkey

Background: Levan is a biopolymer consisting of β–D–fructofuranose units with β(2–6) linkages between fructose rings.

Investigation of the structure and behavior of levan in aqeous environments is necessary to understand its biological activity and its

potential use in various applications such as carbohydrate–derived drug release. The use of different in vivo and in vitro bioactivity

assays fail to relate the chemical structure and conformation of polysaccharides to their observed biological activity. However,

computational simulations provide the opportunity to investigate the behavior of levan chains, and therefore, enable the means to

control its function under various conditions.

Materials-Methods: In the current study, Molecular Dynamics (MD) simulations were used to investigate the static and dynamic

properties of levan chains in water as a function of salinity at 298 and 310 K, representing room and physiological temperatures,

respectively. The simulations were performed for a duration of 5 ns for 9 levan chains each having 12 fructose rings.

Results-Discussion: Investigation of certain static and dynamic properties revealed that the simulations equilibrated around 1 ns

and the levan chains showed Gaussian behavior. At long times, the chains exhibited Fickian behavior and the diffusion coefficients

decerased with incrteasing salt concentration. This behavior was attributed to electrostatic coupling effects. A tendency to

agglomeration was observed in the simulations.

Conclusions: Levan polysaccharides were successfully simulated via Molecular Dynamics simulations. The static properties were not

affected by salt concentration, however, dynamic properties were strongly coupled with salt concentration. Agglomeration of levan

chains were observed, however, longer simulations are needed to quantify this phenomena and its dependence on slat

concentration.

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PP05-14 VISCOELASTIC PROPERTIES OF POLYSACCHARIDE LEVAN VIA NANOINDENTATION

EXPERIMENTS

Brendan Lenz1, Kenneth Noll

1, Deniz Rende

1, Rahmi Ozisik

1, Ebru Toksoy Oner

2

1Department of Materials Science and Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA 2Department of Bioengineering, Marmara University, Istanbul 34722, Turkey

Background: Levan is a naturally occurring polysaccharide that is composed of β–D–fructofuranose units with β(2–6) linkages

between fructose rings. It is synthesized by the action of a secreted levansucrase (EC 2.4.1.10) that converts sucrose into the levan

externally (exopolysaccharide). Levan is a homopolysaccharide that is non–toxic, water soluble, and has anti-tumor activity and low

immunological response. Therefore, levan presents great potential to be used as a novel functional biopolymer in foods, feeds,

cosmetics, pharmaceutical and chemical industries. Despite these favorable properties, levan has a moderately low mechanical

properties and poor film forming capability.

Goal: Investigate the mechanical and viscoelastic properties of levan and the role of mannitol introduced into the fermentation

medium during levan synthesis on its properties.

Materials-Methods: In the current study, the agglomeration behavior of levan in water and in saline solutions was investigated at

298 and 310 K by dynamic light scattering and transmission electron microscopy (TEM). The viscoelastic properties of neat and

oxidized levan films with and without mannitol were studied via nanoindentation experiments in the quasi-static and dynamic

modes.

Results-Discussion: Experiments revealed that addition of mannitol to the fermentation medium substantially increased levan

production rate and the viscoelastic properties. For example, the relaxation coefficients obtained by fitting the loading portion of

the nanoindentation experiments showed a drastic increase with mannitol. The viscoelastic properties also depended on loading

rate, however, this dependency was weaker compared to the effect of mannitol.

Conclusions: The presence of mannitol in the fermentation medium drastically influenced the properties of levan.

119

Mechanical Testing of Adhesives and Adhesion

Theory

120

PP06-01 VISUALIZING THE JKR CONTACT FORMATION AND BREAKAGE USING HIGH SPEED

VIDEO RECORDING

Alexey Tsipenyuk1, Lars Heepe

2, Stanislav N. Gorb

2

1Department of Mechanical Engineering, Technion, Haifa, Israel 2Functional Morphology and Biomechanics, Zoological Institute, Kiel University, Kiel, German

Background-Aim: The JKR model, proposed by Johnson, Kendall and Roberts in 1971, which has been extensively applied to various

bioadhesion systems, is a standard for solving adhesive contact problems. Several experimental studies were performed to verify

the results of the JKR predictions. However, until now, no work has been conducted to monitor the change of the shape and size of

the contact area in real time. The goal for this work was to investigate and provide new information on the structure and dynamic

behavior of the hemispherical probe contacting a flat surface interface, using a high speed contact imaging system.

Materials-Methods: The deformable half-spherical probes were made out of polydimethylsiloxane (PDMS) polymer using a replica

method, from precision plane-convex lenses. Attachment and detachment behaviour was recorded using a high-speed camera

installed on an upright light microscope operated in epi-illumination mode. Video recordings were accompanied by force

measurements. Experiments were performed at different velocities and the video recorded at different optical magnifications.

Results-Discussion: The results obtained showed that with imaging speed of up to 180,000 fps it was possible to resolve the very

moment of detachment. The high-speed image sequences were analysed to quantify the dynamics of the detachment using the

extraction of crack fronts by imposing a simple threshold upon each frame. The local crack propagation velocities were calculated.

This work provided an important experiment for understanding JKR contact theory, which is widely applied to characterize various

biological and biologically-inspired adhesive systems.

This work was performed in the frame of a Short Term Scientific Mission of the COST Action TD0906 (Reference: ECOST-STSM-

TD0906-030713-031885)

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PP06-02 EFFECT OF DETACHMENT SPEED ON THE ADHESION OF SMOOTH FOOTPADS IN

STICK INSECTS

David Labonte, Walter Federle

Department of Zoology, University of Cambridge, Cambridge, UK

Background-Aim: Many insects secrete small amounts of fluid from their adhesive footpads, which may increase adhesion via

capillarity and viscosity. Viscous forces strongly increase with pull-off velocity, potentially hindering locomotion. How does

adhesion of insect pads depend on detachment speed?

Materials-Methods: We measured pull-off forces of individual adhesive pads of live stick insects for different detachment speeds,

using a custom-made 1D force transducer, while filming the contact area with high speed. In order to study the influence of the

secretion volume, experiments were additionally conducted for pads that had been depleted of pad secretion.

Results: Despite a five-hundredfold increase in detachment speed, peak adhesion increased only by less than one order of

magnitude, indicating a weak yet significant effect (ANCOVA, F1,288 = 170; p < 0.001). Adhesion increased for smaller secretion

volumes ( ANCOVA, F8,288 = 2.43; p = 0.015), but this increase was independent of detachment speed (ANCOVA, F8,288 = 1.06; p =

0.39).

Discussion-Conclusion: Our results confirm that the adhesion of insect pads increases with detachment speed. Whilst smaller fluid

volumes increased adhesion, indicating a significant contribution of capillary forces, the velocity-dependence was fluid volume-

independent. This indicates that viscous effects due to the fluid secretion are not the dominant factor driving the velocity-

dependence of adhesion. Instead, the velocity-dependence may be explained by a change in crack-propagation speed, resulting

from viscoelastic effects within the smooth pad’s soft cuticle.

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PP06-03 STICKY PADS - BIOMECHANICS OF FROG ADHESIVE PADS

Thomas Endlein, Diana Samuel, Niall Crawford, Jon Barnes

University of Glasgow, Centre for Cell Engineering, Institute for Cell, Molecular and Systems Biology, Joseph Black Building, University Avenue,

Glasgow G12 8QQ, United Kingdom

Many frogs are able to climb smooth surfaces by using adhesive pads on their feet. This poster highlights four different aspects of

their attachment abilities, which might enable the fabrication of artificial adhesive pads that work in a similar manner.

1) When tree frogs are challenged to cling onto overhanging surfaces, they use opposing feet to create large friction forces. By

doing so, they effectively prevent their pads from peeling which in turn enhances adhesion.

2) Torrent frogs are able to climb in very wet environments. When we compared tree frogs to torrent frogs on wet surfaces, the

torrent frogs showed greater attachment ability. Their superior adhesion is based on various factors, such as having a smaller body

size. They also have a slightly different pad morphology, which possibly allows for faster drainage of water from underneath their

pads.

3) Small tree frogs can hold their entire body weight upside down on a smooth surface, yet even with such sticky pads they can

easily walk on the same surface. To detach their pads quickly and efficiently, they use a peeling mechanism, which results in the

pull-off force being concentrated on a small part of the contact area only.

4) Being sticky means these pads can potentially foul quickly by attracting dirt particles. Here we show that a self-cleaning

mechanism, based on short shear movements of the pads against the substrate combined with a possible ‘ flushing’ effect of the

adhesive fluid, helps to keep the pads clean for good adhesion.

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PP06-04 NOVEL FLOW CHAMBER FOR THE DETERMINATION OF THE ATTACHMENT

STRENGTH OF MARINE ORGANISMS

Simone Dimartino1, Anton Mather

2, Suhas Nawada

1, Meir Haber

3

1Department of Chemical and Process Engineering, University of Canterbury, Christchurch, New Zealand; Biomolecular Interaction Centre,

University of Canterbury, Christchurch, New Zealand 2Department of Biological Sciences, University of Canterbury, Christchurch, New Zealand 3Biota Ltd., Haallon 473/5, Or Akiva 30600, Israel

Background: Bioadhesives secreted by marine macroalgae perform in challenging wet environments. This is especially true for

zygotes, which need to withstand washout and must rapidly secrete a permanent adhesive to secure them to substrates. However,

assessment of zygotes’ adhesive strength is challenging due to small adhesion area and minute quantities of adhesives produced.

Aim: In this study the adhesion strength of Hormosira banksii zygotes is assessed against different surfaces of biomedical relevance,

including a human skin model surface. This model surface simulates in detail both human skin composition and topography.

Materials-Methods: The skin model consists of a blend of cross linked gelatin and lipids, and models are assessed with and without

human skin topography. A drag force is generated on recently settled zygotes using a simple and inexpensive flow channel. The

cells are observed during operation through a Perspex top using a microscope with camera attachment.

Results-Discussion: The flow channel system designed allows testing in situ the adhesion performance of living zygotes. The flow

pattern and streamlines in the flow chamber are simulated using computational fluid dynamics and the drag force acting upon the

zygotes is determined. Cell wash out rate is correlated with drag force, and the adhesion force produced by the natural adhesive is

determined.

Conclusions: This work highlights both the use of a novel flow apparatus to test the adhesion strength of bioadhesives as well as

the potential use of macroalgae inspired adhesives in the biomedical industry.

Part of this work was supported by grant G5RD-CT-2001-00542 from the European Union.

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PP06-05 DETERMINATION OF THE YOUNG´S MODULUS OF THE EPICUTICULA OF THE

SMOOTH ADHESIVE ORGANS OF CARAUSIUS MOROSUS BY TENSILE TESTING

Michael Bennemann1, Stefan Backhaus

1, Ingo Scholz

1, Daesung Park

2, Joachim Mayer

2, Werner Baumgartner

3

1Department Cellular Neurobionics, Institute for Biology II, RWTH Aachen University, Worringerweg 3, 52074 Aachen, Germany 2Central Facility for Electron Microscopy, RWTH Aachen University, Ahornstraße 55, 52074 Aachen, Germany 3Institute of Biomedical Mechatronics, University of Linz, Altenbergerstr. 69, 4040 Linz, Austria

Background-Aim: Adhesive organs of insects like arolia allow these animals to climb on different substrates by creating high

adhesion forces. According to the Dahlquist criterion, arolia must be very soft exhibiting an effective Young's modulus of below 100

kPa to adhere well to different substrates. In previous studies the effective Young´s moduli of adhesive organs were determined

using indentation tests yielding their structure to be very soft indeed. However, arolia show a layered structure, thus the measured

values comprise the effective Young´s moduli of the whole organs.

Materials-Methods: We now developed a new approach to measure the Young´s modulus of the outermost layer of the arolium,

i.e. of the epicuticula, of the stick insect Carausius morosus. Due to the inner fibrous structure of the arolium tensile tests allow the

characterisation of the overlying epicuticula. In our tensile tests arolia of stick insects adhering on a latex membrane were

stretched by stretching the latex membrane while the elongation of the contact area between an arolium and the latex membrane

was recorded. The observed elongations were analysed using finite element simulations.

Results (quantitative and/or statistical data): The simulations yield Young´s moduli of the epicuticula of about a hundred

megapascal.

Discussion-Conclusion: Thus in arolia a very thin layer (~225 nm) of a rather stiff material which is less susceptible to abrasion and

contamination makes contact to the substrates while the inner fibrous structure of arolia is responsible for their softness.

125

PP06-06 CONTACT MECHANICS OF MUSHROOM-SHAPED ADHESIVE MICROSTRUCTURE:

THEORY AND EXPERIMENTS

Lars Heepe1, Giuseppe Carbone

2, Alexander Kovalev

1, Alexander Filippov

3, Stanislav Gorb

1

1Department Functional Morphology and Biomechanics, Zoological Institute of the University of Kiel, Am Botanischen Garten 1–9, Kiel, Germany 2TriboLAB - Dipartimento di Ingegneria Meccanica e Gestionale, Politecnico di Bari, Bari, Italy 3Donetsk Institute for Physics and Engineering of NASU, Donetsk, Ukraine

We studied both experimentally and theoretically the detachment behavior of Mushroom-Shaped Adhesive MicroStructures

(MSAMSs) under different load conditions. Experimental results not only support the homogeneous (optimized) interfacial stress

distribution proposed by recent theoretical model, but also the detachment dynamic is in agreement. The small influence of tilting

on measured pull-off forces is quantitatively well confirmed by numerical and theoretical calculations and was shown to be a direct

consequence of the optimized stress distribution in the contact interface between MSAMS and substrate. The results obtained

allow us to explain the advantage of the widely observed mushroom-shaped contact geometry in nature for long-term and

permanent adhesion

Figure 1

Normalized contact area of three MSAMSs as a function of time. Data were obtained from individual frames of high-speed video

recordings at 5400 frames/s. The very moment of detachment could not be properly resolved as indicated by the sharp transition

(dashed box). The decrease in contact area was almost entirely due to shrinkage without losing contact (see still images)

126

Figure 2

(a-c) MSAMSs in contact with the glass (dark area). (d-f) Color-coded spatiotemporal detachment map. Blue regions detached first,

red regions detached last, and areas sharing the same color detached at the same time. (g-i) Color-coded spatiotemporal crack

velocity map. Scale bar of 10 µm applies to all images.

127

PP06-07 PUSH-OUT BOND STRENGTH OF SELF-ADHESIVE RESIN CEMENTS

Gabriel Furtos1, Bogdan Baldea

2, Luminita Nica

2

1Raluca Ripan Institute of Research in Chemistry, Babes-Bolyai University, Cluj-Napoca, Romania 2Faculty of Dental Medicine, Victor Babes University of Medicine and Pharmacy Timisoara, Romania

Objective: The aim of the present study was to evaluate the retentive strength of two self-adhesive resin cements used for the

cementation of fiber posts into root canals.

Materials-Methods: Twenty extracted non-carious human maxillary central incisors, with fully developed apices extracted for

periodontal reason were used for this study. After endodontic treatment, glass fiber posts were cemented with two types of self-

adhesive resin cements (RelyX U200, 3M ESPE, and Maxcem Elite, Kerr). Four until six sections of 1 mm were cut perpendicular on

the post from each specimen using a diamond saw starting 1 mm coronal from the tip of the post. Post sections were tested on

push out bond strenght using universal testing machine. Mean values of push-out bond strength for each group and root region

(cervical, middle and apical) were calculated. Data were statistically analyzed with one-way ANOVA and Tukey’s test (p<0.05).

Failure modes were evaluated using optical microscopy and scanning electron micrography.

Results: Without being statistically significant, the bond strength of RelyX was higher (8.23±4.46 MPa) when compared to that of

Maxcem Elite (6.52±3.68 MPa). Significant differences (p<0.05) were observed between the apical and cervical regions. More

frequent (>60%) adhesive failures at the resin cement-dentine interface were observed.

Conclusions: The mean push-out bond strength of teeth samples containing RelyX U200 was higher than that observed for Maxcem

Elite. Increases in cement thickness in the cervical third may account for the lower push-out bond strength values obtained in

comparison to the middle and apical thirds in both groups.

Acknowledgements: The authors thank the COST Action TD0906 for meeting support. We are grateful to the National Research Plan

for financial support through the Project no. 165/2012 and no. 189/2012.

128

Molecular Mechanisms of Adhesion

129

PP07-01 BIOADHESION OF A FRESHWATER POLYP

Marcelo Rodrigues, Willi Salvenmoser, Bert Hobmayer, Peter Ladurner

Institute of Zoology and Center of Molecular Bioscience Innsbruck, University of Innsbruck, Technikerstr. 25, A-6020 Innsbruck, Austria

Background: A fundamental challenge in materials science is engineering durable adhesive bonds in a wet environment. Several

organisms are able to solve the problems of underwater adherence, but the nature of the biological molecules involved in adhesion

is not well known. The goal of our project is identifying these molecules involved in biological adhesion using as model organism

the cnidarian Hydra magnipapillata. This species harbours several features that render it as suited to study biological adhesion

including the strong adhesion in wet surfaces in freshwater through basal disc-secreting-mucous cells, the state-of-the-art

methodological toolbox and, the excellent knowledge of the biological features of this organism.

Methods-Results: Essential to our approach is to narrow down the number of genes that are involved in Hydra adhesion. To

achieve this goal, we had, i) characterized the morphology of Hydra basal disc by means of light and electron microscopy; ii) carried

out a differential transcriptome experiment which generated a highly basal disc-specific transcriptome database of which will be

selected about 300 transcripts. This list of candidate transcripts will be analysed by gene expression screen (by in-situ

hybridization), and functionality (by RNA interference). Based on phenotypic validation, genes will be selected to generate

polyclonal basal disc cells specific antibodies.

Conclusion: Using this iterative multidisciplinary approach, the adhesion´s mechanism can be experimentally studied, and

theoretically analysed. Expected outcomes will ultimately to process the appropriate protein/s into useful application that mimics

their adapted functions in the Hydra.

Supported by Marie-Curie FP7-PEOPLE-2013-IEF 626525 and COST Action TD0906

130

PP07-02 CHARACTERISATION OF A NATURAL TEMPORARY ADHESIVE BY FUNCTIONAL

GENOMICS

Lorraine McGill1, James Carolan

2, David Fitzpatrick

2, Thomas Keane

3, Suzanne Jarvis

1

1Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland 2Biology Department, National University of Ireland Maynooth, Maynooth, Co. Kildare, Ireland 3The Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1HH, UK

Background-Aim: The ectoparasite Entobdella soleae attaches to its fish host via a semi-permanent posterior haptor and temporary

secretion of a proteineaous adhesive from specialised glands known as adhesive pads. Amyloid fibrils provide the adhesive and

cohesive strength of this instantly and reversibly acting bioadhesive which is sufficiently strong to withstand the shear forces of

water currents. The aim of this project is to characterise the protein composition of the adhesive using functional genomics.

Materials-Methods: Isolated genomic DNA was sized using flow cytometry and used to generate paired-end and mate-paired

libraries which were sequenced on an Illumnia HiSeq 2000 (GATC Biotech). Genome assembly was conducted and a predicted

protein set was generated using AUGUSTUS. Predictive software identified potential amyloid precursor proteins.

Results (quantitative and / or statistical data): In total 680 million reads were generated and resulted in ~100,000 contigs and our

current assembly has an N50 of 113 bp and 20X coverage of the genome. The 1.3 GB genome is AT rich (>70%) and has a high

incidence of duplication.

Discussion: The E. soleae genome is AT rich, repetitive and large in relation to other platyhelminths. Sequencing coverage was low

but sufficient to generate a predicted protein set. This facilitated the mining for potential amyloid and adhesive proteins. It will

provide a dataset for future direct identification of collected adhesive through high-resolution mass spectrometry.

Conclusion: The E. soleae genome will provide an invaluable resource improving our understanding of the bioadhesives

underpinning this novel and intriguing interaction.

131

Other

132

PP08-01 EFFECTS OF CHRONIC POISONING INDUCED WITH SODIUM NITRATE IN RATS

TESTIS TISSUE WITH TREATMENT

Fatimah Qasim Mohammed1, Nadhem Ahmad Hasan

1, Enas Osamah Huseen

3

1Mosul University, College of Science, Department of Biology, Mosul, Iraq

This study was conducted to investigate the effects of vitamin C and Methylene Blue(MB)on histological changes in adult rats testis

which were treated with Sodium Nitrate(SN).The study included 24 adult white albino male rats aged between (2.5-3monthes)

distributed randomly into four group,each group included 6 animals.The groups were control group, SN group was treated with (

100 mg/kg b.w ) for 60 days, vit C group was treated with vit C ( 50 mg/kg b.w ) after an hour from the treatment with the SN ( 100

mg/kg b.w ) for 60 days and MB group was treated with MB (20 mg/kg b.w ) after an hour from the treatment with SN for 60 days.

The administration of SN ( 100 mg/kg b.w ) daily for 60 days into rats was found to cause congestion in blood vessels between the

seminiferous tubules, sperm absence in several cavities of seminiferous tubules.Additionally hypertrophy in spermatocytes with

presence of degeneration.While it is found hypertrophy in volume of tubules with oedema, degeneration and necrosis in

spermatocytes and sertoli cells,also it is found atrophy in some of tubules in the group treated with vit C after SN.The

administration of MB after SN has lead to shrinkage in volume of tubules, congestion, pyknesis of Sertoli cells nuclei and

accumulation of sperm in some cavities of seminiferous tubules.

133

PP08-02 INFLUENCE OF MOLECULAR WEIGHT ON STRUCTURE AND RHEOLOGICAL

CHARACTERISTICS OF MICROCRYSTALLINE CHITOSAN/POLYACRYLAMIDE

MIXTURES

Katarzyna Lewandowska, Alina Sionkowska

Nicolaus Copernicus University, Faculty of Chemistry, 7 Gagarin Street, 87-100 Toruń, Poland

Aim: The purpose of this study was to evaluate the physico –chemical properties of microcrystalline chitosan differing in molecular

weight with partially hydrolyzed polyacrylamide on the basis of rheological measurements and atomic force microscopy.

Materials-Methods: Flow measurements were carried out on solution of pure polymers: microcrystalline chitosan (MCCh)(with a

degree of deacetylation of 86% and Mv= 5x105 for MCCh I and Mv=9x105 for MCCh II) and polyacrylamide (PAM) (with a degree of

hydrolysis of 1% and Mv= 1x106) and their mixtures with various weight fractions of components. Rheological measurements were

conducted on a rotary viscometer Bohlin Visco 88 with concentric cylinder at different temperatures and shear rates.

The surface morphology of films were analyzed by using a commercial AFM a MultiMode Scanning Probe Microscope Nanoscope

IIIa (Digital Instruments/Veeco Metrology Group, Santa Barbara, CA) operating in the tapping mode in air.

Results-Discussion: The criterion of miscibility of solution mixtures based on the additivity rule of apparent shear viscosity has been

discussed. It was found that studied polymer solutions exhibited the non-Newtonian behaviour with shear-thinning and/or shear-

thickening areas. The positive deviations from the additivity rule have been observed in all cases of investigated mixtures. The

deviations of log ηa decrease with the increase of the temperature. The surface morphology characteristics of MCCh, PAM and

their mixtures films are different. The surface roughness increases in the MCCh/PAM mixtures.

Conclusion: All the above results suggest that modification in properties is a consequence of the strong interaction between the

polymeric components.

134

PP08-03 PREPARATION, STRUCTURAL STUDIES AND THERMAL PROPERTIES OF

COMPOUNDS BASED ON ASYMMETRIC LIGANDS

Imene Mebarka Benguechoua, Djamel Benbertal, Med Taouti

University of Laghouat; Algeria

The main theme of this work is based on the synthesis and characterization of materials based on inorganic ligands. These

compounds can be prepared by methods in solutions at low temperature and that could be used in several industrial and scientific

fields including nonlinear optics, for corrosion inhibiting, but also in medical imaging, biology and catalytic applications. Three

varieties of compounds were synthesized: AMoO4 with A = {Ca, Sr and Ba}, A2MoO4 with A={Ag} that appear by double

decomposition in aqueous solutions, while the third variety A2(MoO4)3 with A = {Al, La and Ce} was synthesized according to a

procedure followed by a heat treatment in order to obtain well-crystallized phases. Their crystal structures were confirmed from

data of X-ray diffraction by the powder and the different software used. The atoms of Ca, Sr, Ba, and La are in coordination 8

belonging to the scheelite structure and atoms of Ag and Al are in coordination 6 belonging to the wolframite structure. The heat

treatment of amorphous phases was possible to obtain well-crystallized phases and also to confirm the thermal stability of these

compounds at high temperatures that can exceed 1000 °C. The use of infrared spectroscopy (FTIR) allowed in a preliminary step to

confirm the existence of molybdats and to assert their wide transparency range extending up to 13μm what makes it possible to

use them in the optic field like laser materials, by doping them with luminescent elements

135

PP08-04 NEW MATERIAL FOR BIOMEDICAL APPLICATION MADE OF FISH COLLAGEN

Alina Sionkowska, Justyna Kozlowska

Faculty of Chemistry, Nicolaus Copernicus University, Poland

Background: Collagen, a major structural component of extracellular matrices (ECM), is known for its high cell adherence capacity.

The aim of this study was to determine the surface properties of thin collagen matrices - an excellent substrate for the culture of

many different cell types.

Materials-Methods: Collagen was isolated from fish scale of Esox lucius, a calcified tissue, through demineralization following

acetic acid treatment. The surface properties of collagen films were investigated using the technique of Atomic Force Microscopy

and by contact angle measurements using diiodometan and glycerol. The surface free energy was calculated using Owen Wendt

method.

Results: Using AFM technique we observe the fibrils formation. diameter of fibril is 55 nm. The RMS (root-mean-square) surface

roughness of collagen is 7,5 nm. The value of surface free energy is 37.54 mN/m and the polar component of surface free energy is

13.18 mN/m.

Discussion: Surface physicochemical properties play a significant role in biocompatibility since they govern cellular adhesion,

proliferation and performances. We obtained a roughness film characterized by low polar component of the surface free energy.

Conclusion: Results indicate that the topography of fish collagen film is suitable for interactions of cells with the material`s surface.

Financial support from the National Science Centre (NCN, Poland) Grant No. UMO-2012/05/N/ST8/02283 is gratefully

acknowledged.

136

PP08-05 SYNTHESIS OF HIGHER HOMOLOGOUS OF MOLECULES WITH BIOLOGICAL

INTERESTS

Fatima Rouda, Brahim Sabir, Abdelmejid Bahloul

Laboratory of Biomolecular and Organic Synthesis, Faculty of Sciences Ben Sik University

This is a part of the undertaken work in our laboratory [2] for the synthesis of heterocyclic components via dipolar cycloaddition-1,

3 with Diarynitilimine (DANI) [1]; which have revealed biological activities [3].

Thus, the objective of this paper is focused on the synthesis of higher homologue of molecules with biological interest.

As part of this work, we worked initially on cloves in carrying out the extraction of essential oil of two main components which are

eugenol and acetyleugenol.

In a second step, we performed the reaction of 1,3 dipolar cycloaddition with diarylnitrilimines (DANI) on eugenol and

acétyeugénol as dipolarophiles to synthesize new membered heterocyclic compounds such as pyrazolines or isoxazolines, which

were characterized by IR, 1H and 13C-NMR, and we studied the regiochemistry, and the stereochemistry of the cycloaddition

reaction.

References:

[1]R.Huisgen,Angew,Chim.1963,75,742-754;,Angew,Chim.INTERNE.ED.L’Agleterre.1963,14,633.

[2]A.Bahloul Doctorat és-sciences physiques.Faculté des sciences Ben M’sik.Casablanca1998

[3] A.Sban, A.Bahloul, M.Berrada, S.Kitane, K.Bougrin, M.Soufiaoui, Cycloaddition dipolaire-1,3,:Etude de la régioselectivité des nitrilimines

disubstituées et des arylnitriloxides avec l’isochroméne, journal de la Société chimique de Tunisie 4 (11) (2002) 1403-1049

137

PP08-06 NANOFUR: FROM HIERARCHICAL GECKO-BASED ADHESION TO FLUID ADHESION

Maryna N Kavalenka, Michael Röhrig, Andreas Hopf, Marc Schneider, Matthias Worgull, Hendrik Hölscher

Karlsruhe Institute of Technology, Institute of Microstructure Technology, Eggenstein-Leopoldshafen, Germany

Background-Aim: Fascinating properties of many surfaces occurring in nature originate from nano- and micro-hairs covering them.

Materials-Methods: Inspired by natural surfaces, we introduce a highly scalable low-cost fabrication technique for producing a

dense fur of high aspect ratio nanohairs out of polymer surfaces. This hot pulling technique is a modified hot embossing process in

which softened polymer is locally elongated with a heated sandblasted steel plate, creating densely packed nanohairs on the

polymer surface.

Results (quantitative and / or statistical data): Combination of hot embossing and hot pulling was used to fabricate a three-fold

hierarchy inspired by the adhesive toes of geckos. Additionally, by changing the hot pulling fabrication parameters the density and

surface of the nanofur can be varied, resulting in different water and oil droplet adhesion.

Discussion: Fabricated three-fold structures exhibit an improved adhesion/preload ratio and large work of adhesion. Moreover, the

as-fabricated nanofur surface is both superhydrophobic and superoleophilic and can selectively absorb oil from the oil/water

mixture. Plasma-treated nanofur surface exhibits low adhesion of oil underwater and is underwater superoleophobic.

Conclusion: Hot pulling technique was used to fabricate adhesive structures inspired by the toes of geckos, and structures with

different water and oil droplet adhesion. High and low adhesion of droplets is advantageous for different applications including

microfluidic devices, self-cleaning surfaces and materials for oil/water separation.

138

PP08-07 IN VITRO ANTIOXIDANT ACTIVITIES BY PHENOLIC EXTRACTS OF ZYGOPHYLLUM

ALBUM FROM SOUTH ALGERIA

Majda Benguechoua, Samira Nia, Khedidja Benarous, Ihsen Khecheba, Mohamed Yousfi

University of Laghouat

Phenolic compounds are bioactive molecules exhibiting a lot of scientific attention due to their multiple biological activities. In this

study we are interested in phenolic extracts of zygophyllum album from south algeria and in vitro study of their antioxidant.

The first part of this study concerns the extraction and quantification of phenolic compounds, flavonoïds and tannins with different

systems: Methanol / Eau; Acétone /Eau.

From the results, zygophyllum album is rich in total phenols ( ranging from 3.022±0.31 mg/g to 5.52±0.352mg/g in ethyl acetat

fractions and varies from 6.353±1.368 mg/g to 8.205± 0.844mg/g in the butanol fractions of gallic acid equivalent), they are also

rich in flavonoids (ranging from 0.258±0.129 mg/g to 0.395±0.197mg/g in ethyl acetat fractions and varies from 1.05±0.52 mg/g to

1.213± 0.0.617mg/g in the butanol fractions of quercetine equivalent ) and also rich in tannins ( ranging from 0.200±0.083 mg/g to

0.0.221±0.022mg/g in ethyl acetat fractions and varies from 1.05±0.108 mg/g to 1.99± 0.177mg/g in the butanol fractions of

catechine equivalent). In the second part, we studied antioxidant and radical scavenging activities using DPPH radical scavenging

and phosphomolybdenum methods. Results have shown that our extracts posses a moderate antioxidant activity in comparison

with standards

139

PP08-08 QUIET MALE BEETLES ATTACH STRONGLY TO FEMALE’S BACK

Dagmar Voigt1, Alexey Tsipenyuk

2, Michael Varenberg

2

1Institute for Botany, Technische Universitaet Dresden, D-01062 Dresden, Germany 2Department of Mechanical Engineering, Technion, Technion City, 32000 Haifa, Israel

Background-Aim: Tarsal adhesive setae of leaf beetles attract scientists’ interest since several decades. In particular, male-specific

structures, bearing discoid terminals, successfully inspired effective biomimetic adhesives. However, attachment forces of male

beetles have been mostly measured on artificial and few plant surfaces. The natural counterparts (female elytra), which are met

during copulation, were not considered so far.

Materials-Methods: Attachment abilities of living rosemary beetles (Chrysolina americana L.) were measured using the custom

tribometer capable of evaluating both adhesion and friction forces. A living female was firmly attached ventrally to a measuring

lever, and a living male dorsally to a moving table. Male head and legs were freely movable. Feet were brought in contact with

female elytra, and pulled-off while measuring attachment forces.

Results-Discussion: Forces depended on males’ motion, body and leg posture. The more tightly and quietly sat males on females’

elytra, the stronger they kept attached to them.

While pulling-off the motionless firmly attached males, adhesion force was estimated to be of up to 60 mN, corresponding to about

90 times the male body’s weight. These values support previous studies on maximum attachment forces of beetles caused by the

specifically evolved discoid-shaped male adhesive setae.

Our study provides further insight into the male leaf beetles' attachment to females' back during copulation and long-term mate

guarding, which is a fascinating but still not fully understood phenomenon. The results obtained may also be useful in designing

future biomimetic adhesives.

Financial support: ECOST-STSM-TD0906-171113-036395 „In situ SEM of biological adhesive contacts in action”

140

PP08-09 INSIGHTS INTO LONGHORN BEETLE ATTACHMENT

Dagmar Voigt1, 3

, Takuma Takanashi2, Kazuko Tsuchihara

2, Kenichi Yazaki

2, Katsuhiko Kuroda

2, Remi Tsubaki

2, Naoe Hosoda

3

1Institute for Botany, Technische Universitaet Dresden, Dresden, Germany 2Forestry and Forest Products Research Institute, Tsukuba, Ibaraki, Japan 3Interconnect Design Group, Hybrid Materials Unit, National Institute for Materials Science, Tsukuba, Ibaraki, Japan

Background-Aim: Most longhorn beetles impress with their considerable body size, long antennae, long legs bearing expanded

hairy tarsomeres and well developed paired claws. They have been observed to attach strongly to various surfaces. However, host

substrates and the attachment system were not considered in detailed studies so far. Exemplarily, plant surfaces and tarsi of

Japanese pine sawyer beetle (Monochamus alternatus Hope) were visualized in order to shed light on longhorn beetle attachment.

Materials-Methods: Light microscopic observations, cryo-scanning electron microscopic studies and measurements of free surface

energy of host substrates were carried out. Additionally, traction forces of male and female longhorn beetles on glass were

measured and set into the context.

Results-Discussion: Monochamus alternatus may attach to a range of surfaces, including pine needles, twig and stem bark as well

as female elytrae, which all vary in texture and free surface energy. Five tarsomeres are densely covered with in total about 23600

ribbon-shaped adhesive setae bearing oval spatula-shaped, tapered terminals. Terminals are dorsally equipped with 5 to 10

setulae. In contrast to numerous leaf beetles and ladybirds, no sexual dimorphism in shape of adhesive setae is observed. 3rd

tarsomere in both sexes appears clearly belobed. Adhesive setae occur anisotropically arranged. On flat glass, up to 102 mN pulling

force is generated, whereas up to 206 mN on glass tubes - considerably stronger pulling forces compared to those of other insects.

We suppose an adaptation of the tarsal attachment system of Monochamus alternatus to tubular structures.

Financial support: ECOST-STSM-TD0906-010314-040971 and JSPS Grant-in-Aid for Scientific Research on Innovative Areas Grant

Number 24120005 & 24120006

141

PP08-10 ASSESSMENT OF ANTIOXIDANT POTENTIAL, TOTAL PHENOLICS AND FLAVONOIDS

OF DIFFERENT SOLVENT FRACTIONS OF OUDNEYA AFRICANA AREAL PARTS

Samira Nia, Madjda Benguechoua, Khedidja Benarous, Mohammed Yousfi

Laboratoire des sciences fondamentales –university of amar télidji- BP 37G Laghouat Algeria

Objective: This study was conducted to investigate the antioxidant potential of aqueous acetone extract and its derived fractions

(dichloromethane, ethyl acetate and n-butanol) of Oudneya africana medicinal plant belonging to the brassicaceae family. This

specie is a desert plant found in Algerian Sahara and used in folk medicine.

Methods: Dried powder of the areal parts of oudneya africana was extracted with aqueous acetone (7/3 v/v) and the resultant was

fractionated with solvents having escalating polarity; dichloromethane, ethyl acetate and n-butanol. Total phenolic and total

flavonoid contents were estimated for the various fractions. These fractions were also subjected to in vitro assay to estimate the

scavenging activity for 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS).

Results-Discussion: The analysis results of phenolic extracts show clearly a high content of phenolic compounds in investigated

plant ranging from 3.34±0.28 to 17.52±0.68 mg/g of gallic acid equivalent and the total flavonoids content lie between 0.14±0.02

and 5.16±0.14 mg / g of quercitin equivalent, the n-butanol fraction possessed high amount of phenolics and flavonoids compared

with other fractions.

The oudneya africana extracts showed a pronounced scavenging activity on ABTS with IC50 values varied from 0.658±0.025 mg/ml

to 1.526±0.033 mg/ml where the ethyl acetate fraction present the best results comparatively to antioxidants taken as references.

Conclusion: These results are preliminary, for a better understanding of potential antioxidant; it would be interesting to

characterize the reaction intermediates and products. To do this, it is necessary to separate these compounds by chromatography

and to identify them.

142

PP08-11 FACILE AND STABLE GOLD NANOPARTICLE IMMOBILIZATION ON SURFACES

THROUGH MUSSEL ADHESIVE POLYPEPTIDES

Maya Bernmalm1, Mattias Berglin

2

1SP Technical Research Institute of Sweden, Chemistry, Materials and Surfaces, Borås, Sweden 2Chemistry and Molecular Biology, Gothenburg University, Gothenburg, Sweden

Gold nanoparticles (Au-NPs) have unique electronic, optical and catalytic properties. The integration of Au-NPs with well-defined

spatial distribution into thin films is particularly important for various applications, for example in tissue engineering, biological

sensing and in the preparation of electronic nanodevices. The general immobilization protocol has included the use of silanes and

dithiols onto flat glass and gold surfaces respectively. Currently the immobilization protocols on polymer surfaces are hampered by

either selecting the appropriate polymer or includes several modification steps including grafting of Au binding moieties. In this

study we evaluated the use of the mussel mytilus edulis foot protein 1 (mefp-1) as adhesive precursor for immobilization of Au-NPs

onto, glass, gold (Au) and polymer surfaces. The kinetic of Au-NPs immobilization was followed by Quartz crystal microbalance with

dissipation (QCM-D), the degree of immobilization (coverage) and spatial distribution was detected by scanning electron

microscopy (SEM). Stability of the immobilized Au-NPs on the surfaces was evaluated in high pH as well as treatment in protein

solution (fibrinogen). Mefp-1 demonstrates slightly slower binding kinetics but similar binding capacity compared with a

polycationic polypeptide (polylysine) on gold and glass surfaces. Mefp-1 is able to bind Au-NPs while polylysine is not able to bind

any Au-NPs on the evaluated polymer surfaces. Au-NPs immobilized on the mefp-1 layer show superior stability in both high pH and

treatment with fibrinogen compared with polylysine. The use of marine polypeptides opens up the possibility to bind Au-NPs on

complex geometries such as tubings and porous polymeric structures.

143

PP08-12 SCREENING OF QUORUM SENSING SIGNAL MOLECULES AND BIOFILM FORMATION

IN ESBL PRODUCING KLEBSIELLA SPP.

Demet Erdönmez1, Abbas Yousefi Rad

3, Nilüfer Aksöz

2

1Institute of Science, Department of Biology, Hacettepe University, Ankara 06800, Turkey 2Faculty of Science, Department of Biology, Hacettepe University, Ankara 06800, Turkey 3Koru Hospitals, Coordinators of The Clinical Laboratory, Ankara,06520,Turkey

Background: The extended-spectrum beta-lactamase (ESBL)-producing Klebsiella spp. cause worldwide problems in intensive care

units.Our study aimed to determinated Quorum sensing signal molecules from ESBL-producing Klebsiella spp. and biofilm

formation.Quorum sensing is a bacterial cell-cell communication process that relies on the production and detection of

extracellular signal molecules called autoinducers. Just as in many pathogens, quorum sensing system of Klebsiella spp. is

responsible for expression of virulence genes.This regulated expression of virulence genes is thought to give the bacteria selective

advantages over host defenses which is important for the pathogenesis of the organism.

Materials-Methods: 20 strains of ESBL positive Klebsiella spp. from different clinical specimens were identified by conventional

methods.ESBL producing Klebsiella spp. strains were detected by using “double disc sinergy method”.QS signal molecules were

identified by cross-validation test with using C.violaceum 026 and A.tumefaciens NTL1 bioreporter strains. QS signal molecules

combined with the biosensor strains were separated thin layer chromatography (TLC) from each other.

Results: According to results,it was found out that 15 of the Klebsiella spp strains produced the following miscellanous QS signal

molecules in varying amounts: C6-HSL and C8-HSL.

Conclusion: Using more than one strain biosensor at cross-validation test is a useful strategy to detect signaling molecules.The

variations in the amounts and types of signal molecules were observed to affect virulence properties.We were demonsrated clinical

isolate, Klebsiella spp were produced between C6-HSL and C8-AHL. This research will ensure better understanding of QS activity

of Klebsiella spp.and may enable the anti-QS as one of the alternative anti-infective therapy.

144

INDEX

145

A Abe Koji 107

Abram Florence 59

Aksöz Nilüfer 143

Albakri Budr 96

Albert Klaus 113

Alcarva Catarina 58

Ali Dhinojwala 13, 14

Ana I. Neto 44

Arai Ryoichi 107

Arga K. Yalcin 54, 117

Ashton Nicholas N. 66

Ateş Özlem 85

B Backhaus Stefan 124

Baer Alexander 55

Bahadori Fatemeh 83

Bahloul Abdelmejid 136

Baldea Bogdan 127

Barhemat Samira 50

Barnes W. Jon. P. 63, 93, 94, 116, 122

Bauer Ulrike 68

Baumgartner Werner 124

Baysal Nihat 54, 117

Belma Saldamli 42

Benarous Khedidja 138, 141

Benbertal Djamel 134

Benguechoua Imene Mebarka 134

Benguechoua Majda 138, 141

Bennemann Michael 124

Berg Albrecht 79

Berglin Mattias 142

Bernmalm Maya 142

Betz Oliver 103

Blasco Pilar Carbonell 77

Bloom Orahn Preiss 86

Bohn Holger Florian 68

Bosia Federico 62

Bramson Nadav 86

Brely Lucas 62

Bucak Seyda 54, 117

Busqué Felix 45, 74

Butt Hans Jürgen 64, 93, 94

C Carbone Giuseppe 125

Carolan James 130

Cem Gulcin 54, 117

Ceylan Hakan 72, 84

Coelho Ana Varela 59

Cordeiro Carlos 58, 115

Coskunkan Binnaz 54, 117

Crawford Niall 63, 122

D Da Costa Gonçalo Conde 58, 115

Da Silva Cristina Ribeiro 115

Dana Aykutlu 84

Das Saurabh 60

De Weerdt Cecile Van 52

Del Campo Aránzazu 73, 93, 94, 100

Demeuldre Mélanie 56, 104

Dimartino Simone 53, 123

Dirk Drotlef 3 8

E Ederth Thomas 50

Elena Bellido 45

Elisa Martinelli 25, 26

Elise Hennebert 17, 18

Endlein Thomas 63, 116, 122

Erdönmez Demet 90, 91, 143

Erginer Merve 48

Erkan Kübra 90, 91

Eroğlu Mehmet S. 48, 83, 85

F Federle Walter 68, 103, 121

Filippov Alexander 125

Fitzpatrick David 130

Flammang Patrick 52, 56, 104, 111

Furtos Gabriel 127

G Garcia Beatriz 74

Giancarlo Galli 26

Goetzke Hanns Hagen 103

Gorb Stanislav N. 65, 87, 120, 125

Gouveia Analuce 58

Grunwald Ingo 59

Guerette Paul 51

Guler Mustafa Ozgur 72, 84

Gulsuner Hilal Unal 72

Guo Shifeng 70

Gylys Maryte 60

146

H Haber Meir 80, 123

Hachisu Masakazu 109

Harrington Matthew J. 55

Hasan Nadhem Ahmad 132

Havazelet Bianco Peled 39

Heepe Lars 87, 120, 125

Hennebert Elise 52, 56, 104, 111

Herbert Waite 22

Hernando Jordi 74

Hirabayashi Kimio 107

Hobmayer Bert 129

Hölscher Hendrik 96, 137

Hoon Shawn 51

Hopf Andreas 137

Hosoda Naoe 65, 140

Huseen Enas Osamah 132

Hwang Dong Soo 61

I Israelachvili Jacob N. 60

Iturri Jagoba 93, 94

J Janczewski Dominik 70

Jarvis Suzanne 130

Jarvis Suzi P. 112

João F. Mano 44

Jon Barnes 38

Jonker Jaimie Leigh 59

Jordi Faraudo 45

K Kappl Michael 64, 93, 94

Katırcıoğlu Hikmet 91

Kaufman Yair 60

Kavalenka Maryna N. 137

Kaveh Farzaneh 64

Keane Thomas 130

Kei Kamino 35

Kenneth J. Moise 37

Khecheba Ihsen 138

Kim Byung Chun 98

Kiwi John 76

Kizilkan Emre 87

Klaus Rischka 42

Kocabey Samet 72

Kovalev Alexander 125

Kozlowska Justyna 135

Kuroda Katsuhiko 140

L Labonte David 121

Ladurner Peter 56, 104, 105, 106, 111, 129

Lee Serina Siew Chen 70

Leite Álvaro J. 85

Lengerer Birgit 104, 105, 106, 111

Lenz Brendan 118

Lewandowska Katarzyna 133

Lipik Vitali 97

Lir Irina 80

Lovepreet Mann

37

M Maldonado Barbara 52

Mano João F. 85

Marco d’Ischia 19, 20

Maria Kozielec 42

Marleen Kamperman 15, 16

Martinez Jose Miguel Martin 77

Mather Anton 123

Mayer Georg 55

Mayer Joachim 124

McGill Lorraine 130

Mcquillan A. James 53

Metin Sitti 40

Michael Mularczyk 42

Miller Dusty R. 60

Mireia Guardingo 45

Miserez Ali 51, 97

Mohammed Fatimah Qasim 132

Molina Daniel Ruiz 45, 74

Mutlu Esra C. 48, 83

N Natália L. Vanconcelos 44

Nawada Suhas 123

Nia Samira 138, 141

Nica Luminita 127

Nick Aldred 23, 24

Noll Kenneth 118

Nomura Takaomi 107, 109

Nugraha Roni 50

O Ohkawa Kousaku 107, 109

Oner Ebru Toksoy 48, 54, 79, 83, 85, 117, 118

Ozisik Rahmi 54, 117, 118

147

P Paez Julieta 73, 100

Park Daesung 124

Park Hyojung 98

Paul Molino 27, 28

Peter Ladurner 36

Phillip B. Messersmith 29, 30

Pjeta Robert 105, 106

Poseu Javier Saiz 74

Power Anne Marie 59

Pugno Nicola M. 62

Puniredd Reddy Sreenivasa 70

R Rachedi Fahima Ali 95

Rad Abbas Yousefi 143

Ramesha Papanna 37

Rende Deniz 54, 117, 118

Ribeiro Raquel Mesquita 58, 115

Riehle Mathis 63, 116

Robert Sader 42

Rodrigues Marcelo 105, 106, 111, 129

Röhrig Michael 137

Ronit Bitton 39

Rothbächer Ute 102

Rouda Fatima 136

Russell J. Stewart 37

S Sabir Brahim 136

Sağlam Necdet 90, 91

Salibasic Alma 112

Salvenmoser Willi 105, 106, 111, 129

Samuel Diana 116, 122

Sang Byoung In 98

Santos Romana 58, 115

Sara M. Oliveira 44

Sarbjit Kaur 37

Sarilmiser Hande Kazak 83

Savory David 53

Schmidt Stephan 55

Schnabelrauch Matthias 79

Schneider Marc 96, 137

Scholz Ingo 124

Sedó Josep 45, 74

Semprebon Ciro 64

Seow Yiqi 51

Serrano Cristina 73

Shahram Ghanaati 42

Shamchi Minoo Pourhassan 91

Silva Joana M. 85

Simões Nelson 58

Sionkowska Alina 133, 135

Song In Wong 98

Stahl Beate 113

Stewart Russell J. 66

Sutanto Clarinda 51

Syurik Julia 96

T Takanashi Takuma 140

Taouti Med 134

Tekinay Ayse Begum 72, 84

Teo Serena Lay Ming 70

Tomer Guy 86

Toubarro Duarte 58

Tsipenyuk Alexey 47, 120, 139

Tsubaki Remi 140

Tsuchihara Kazuko 140

Tsukada Masuhiro 107

Turgut Deniz 54, 117

U Urel Mustafa 84

Ustahüseyin Oya 100

V Vancso Julius Gyula 70

Varenberg Michael 47, 139

Veikko Sariola 40

Verdaguer Albert 45

Voigt Dagmar 139, 140

W Waite J. Herbert 21, 60

Walter Federle 38

Wattiez R. 56

Willi Schwotzer 31, 32

Wolf Johanna 96

Wong Fong T. 51

Worgull Matthias 137

Wunderer Julia 105, 106

Wyrwa Ralf 79

X Xue Longjian 93, 94

148

Y Yandi Wetra 50

Yazaki Kenichi 140

Yoav Rozen 39

Yousfi Mohammed 138, 141

Z Zhang Lihong 97

Zhu Xiaoying 70

Zwiener Christian 113

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