THE BIOMANUFACTURING PROGRAMME AT LEIRIA - WordPress… · 2013-07-20 · www. cdr-sp.ipleiria.pt...
Transcript of THE BIOMANUFACTURING PROGRAMME AT LEIRIA - WordPress… · 2013-07-20 · www. cdr-sp.ipleiria.pt...
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Paulo Jorge Bártolo
THE BIOMANUFACTURING PROGRAMME AT LEIRIA
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Regional Innovation Scoreboard (2012)
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The Centre is located at Marinha Grande in a two floor building of 3320 m2.
The Centre for Rapid and Sustainable Product Development was created in May, 17, 2007 by the President of the Polytechnic Institute of Leiria. It is a Research Centre of the Polytechnic Institute of Leiria (IPL) rated as excellent by the most recent evaluation of Research Units carried out by the Portuguese Foundation for Science and Technology (FCT).
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FUTURE BUILDING
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KEY RESEARCH THEMES:
Advanced materials – materials with novel functionalities enhancing
new manufacturing technologies and processes and the opportunities to manufacture entirely new materials-based technologies and products. SMART MATERIALS, SUSTAINABLE POLYMERS, FUNCTIONAL GRADED STRUCTURES
Emerging technologies – production technologies to exploit the potential of emerging technologies (in particular bio- and multi-scale technologies); leveraging simulation and modelling techniques to address manufacturing challenges; flexible, rapidly responsive production systems for customized manufacturing.
Sustainable manufacturing and manufacturing of green technologies
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GLOBAL FUNDING
0
1000000
2000000
3000000
4000000
5000000
6000000
7000000
8000000
9000000
10000000 2013
2012 2011
2010
2009
2008
2007
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Martinho, P., Bártolo, P.J., Pouzada, A., Rapid Prototyping Journal, 5, 71, 2009
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Biomanufacturing - the use of additive technologies, biodegradable and biocompatible materials, cells, growth factors, etc., to produce biological structures for tissue engineering applications
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BIOMEDICAL IMPLANTS I
Medical implants are devices placed either inside or on the surface of the body to accomplish some particular function, such as to replace, assist or enhance the functionality of some biological structure(s).
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BIOMEDICAL IMPLANTS II
BIO
MED
ICA
L IM
PLA
NTS
Body External implants
Body internal permanent implants
Body internal temporary implants
Hip implants Knee implants
Retinal implants Dental implants
Spinal fusion constructs
Scaffolds Degradable screw and plates
Drug delivery systems Spinal fusion constructs
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BIOMEDICAL IMPLANTS III
Body External implants
Hand
Eye
Foot
Leg
Arm
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BIOMEDICAL IMPLANTS IV
Body External implants
Manabu OKUI – CDRSP & Tokyo Institute of Technology
WALKING INTEGRATIVE REHABILITATION DEVICE
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BIOMEDICAL IMPLANTS V
Body Internal permanent implants
IBEROAMERICAN NETWORK ON BIOMANUFACTURING : MATERIALS, PROCESSES AND SIMULATION (BIOFAB) - ~200 researchers - 18 Universities and Research
Institutes - Collaborative research - Courses - Training
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BIOMEDICAL IMPLANTS VI
Body Internal permanent implants
Bucomaxillofacial prosthesis. Beyond aesthetic purposes, this prosthesis showed functional results, improving patient speech and feeding.
Implants can be functional or cosmetic. Source: CTI, Brazil
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BIOMEDICAL IMPLANTS VII
Naso-orbitary reconstruction. Tumor resection - esthesioneuroblastoma
Surgery time reduced in about 40% Better quality of the surgery due to the fit and shape of the implant
Body Internal permanent implants
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BIOMEDICAL IMPLANTS VIII
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BIOMEDICAL IMPLANTS IX
Biocompatibility. Both raw and processed materials should interact positively with the host environment without eliciting adverse host tissue responses. Biodegradability. Scaffolds must degrade into non-toxic products with a controlled degradation rate that matches the regeneration rate of the native tissue.
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BIODEGRADABLE IMPLANTS X
Source: Holzapfel et al, Advanced Drug
Delivery Reviews, 2012
Appropriate porosity, pore size and pore shape. Generally, a high level of porosity is required (> 90%) because it increases the surface area, enabling high cell seeding efficiency, migration and proliferation, as well as neovascularisation. Pore interconnectivity (100% interconnected network of internal channels are required) is also a critical parameter in terms of cell viability and tissue regeneration, maximizing the diffusion and exchange of nutrients and the eliminations of waste.
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BIODEGRADABLE IMPLANTS XI
Source: Zhang et al, Advanced Drug Delivery Reviews, 2012
Bioactive. Scaffolds should be bioactive, promoting and guiding cell proliferation, differentiation and tissue growth. This can be achieved by adding growth factors and functionalizing the scaffold with proteins or adhesion-specific peptide sequences.
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BIOMEDICAL IMPLANTS XII
Cells
Signals Scaffold
Cell-scaffold interactions
Cell-cell interactions
Tissue Engineering main pillars:
Physicochemical interaction (shear, pH, O2, CO2, temperature
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BIOMEDICAL IMPLANTS XIII
Hydrogels
Polymers Hydrogels
Polymers Polymer/ceramic
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BIOMANUFACTURING @ CDRSP I
MATERIAL CHARACTERISATION MANUFACTURING PROCESSES
COMPUTER MODELLING & SIMULATION
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BIOMANUFACTURING @ CDRSP II
COMPUTER MODELLING & SIMULATION 4D Modelling Approach
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BIOMANUFACTURING @ CDRSP III
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BIOMANUFACTURING @ CDRSP IV
ADDITIVE PROCESSES NON-ADDITIVE
PROCESSES
HYBRID PROCESSES
EXTRUSION-BASED PROCESSES
STEREOLITHOGRAPHIC PROCESSES
ELECTROSPINNING
PRINTING PROCESSES
PLASMA TREATMENT/COATING
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BIOMANUFACTURING @ CDRSP VI
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BIOMANUFACTURING @ CDRSP VII
Melt-electrospun sPCL189 at a temperature of 160 ◦C (Vapp =25 kV, EFR = 3.75±0.08 mL h−1): (a) focused deposition by ‘static’ melt-ES; (b) uniform 3D fibre collection by computer-aided melt-ES (extruder translation velocity 500 mm min−1).
SEMmicrograph of melt-electrospun scaffolds produced with Vapp =25 kV and SRS = 29.0 rpm: (a) linear PCL produced with Tproc =160 ◦C; (b) sPCL64 produced with Tproc =125 ◦C; (c) sPCL189 produced with Tproc =160 ◦C; (d) sPCL189 melt-electrospun fibres collected in a coiled form following the deposition pattern (Tproc =160 ◦C) – the arrow indicates the deposition direction. Inset images show details of the fibre surface topography obtained; scale bar 10 μm.
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VAT-PHOTOPOLYMERISATION I
PEGDMA Dextran PEGDMA/HA PEGDMA/Dextran UP systems
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VAT-PHOTOPOLYMERISATION II
PEG
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VAT-PHOTOPOLYMERISATION III
PEG
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VAT-PHOTOPOLYMERISATION IV
Ethylene glycol dimethacrylate
Triethylene glycol dimethacrylate
The HEMA was copolymerized with 1-vinyl-2-pyrrolidinone using two differents crosslinks agents
1-Vinyl-2-Pyrrolidinone HEMA
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VAT-PHOTOPOLYMERISATION V
HEMA with 0.5wt% of PI
HEMA with 1wt% of PI HEMA with 2wt% of PI
HEMA with 3wt% PI
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VAT-PHOTOPOLYMERISATION VI
Irradiation time (s)
No
rma
lize
d w
ate
r a
bso
rptio
n
0,0
0,2
0,4
0,6
0,8
1,0
1,2
1,4
1,6
1,8
2,0
15 25 35 45 60
T= 15 s T= 60 s
PEGDA
Dextran
Influence of the irradiation time on the water absorption of PEGDMA constructs built by STLG system (left). Macroscopic images of the constructs produced using 15s and 60s of irradiation time, after immersion into distilled water during 24 hours (right). Scale bar represents 0.5mm.
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VAT-PHOTOPOLYMERISATION VII
Energy (KeV)
0 1 2 3 4 5 6 7 8
Counts
0
2000
4000
6000
8000
10000
12000
14000
16000
Energy (KeV)
0 1 2 3 4 5 6 7 8
Counts
0
2000
4000
6000
8000
10000
12000
14000
16000
Ca
Ca
P
C
O
a) b)
EDS analysis and SEM micrographs of the PEGDMA (a) and PEGDMA/HA hydrogel scaffolds (b).
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VAT-PHOTOPOLYMERISATION VIII
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VAT-PHOTOPOLYMERISATION IX
Curing time (s)
0 100 200 300 400 500
Fra
cti
on
al co
nvers
ion
0.0
0.2
0.4
0.6
0.8
1.0
50 wt% of styrene
60 wt% of styrene
70 wt% of styrene
37 wt% of styrene
Coral-like structure
Flake-and-pore structure
Flake-type structure
Curing time (s)
0 100 200 300 400 500
Fra
cti
on
al
co
nve
rsio
n
0.0
0.2
0.4
0.6
0.8
1.0
70 wt% of styrene
37 wt% of styrene
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VAT-PHOTOPOLYMERISATION XI
COMPOSITE STEREOLITHOGRAPHY Magnetic field
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VAT-PHOTOPOLYMERISATION X
Cláudio Ferreira, MSc work
a) b)
c)
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VAT-PHOTOPOLYMERISATION XIII
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EXTRUSION-BASED PROCESSES I
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EXTRUSION-BASED PROCESSES II
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EXTRUSION-BASED PROCESSES III
PCL PCL/PLA PCL/HA PCL/TCP PCL/graphene PCL/PHB PLA Alginate Alginate/aloe vera Alginate/agar Alginate/chitosane
Material characterisation: DSC, FTIR, NMR, GPC, AFM
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EXTRUSION-BASED PROCESSES IV
FIBROBLASTS
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EXTRUSION-BASED PROCESSES V
Saos-2 cells
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EXTRUSION-BASED PROCESSES VI
Human mesenchymal stem cells (hMSCs)
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EXTRUSION-BASED PROCESSES VII
Tendogenesis/
LigamentogenesisMarrow Stroma
Transitory
FibroblastTransitory
Stromal Cell
Osteogenesis Chondrogenesis Myogenesis
MSC Proliferation
Transitory
Osteoblast
Transitory
Chondrocyte Myoblast
Mesenchymal Stem Cell (MSC)
Myoblast Fusion
Unique
Micro-niche
ChondrocyteOsteoblast
Other
Bo
ne
Ma
rro
w/P
eri
os
teu
mM
es
en
ch
ym
al
Tis
su
e
Proliferation
Commitment
Lineage
Progression
Differentiation
Maturation
OsteocyteHypertrophic
ChondrocyteMyotube
Stromal
Cells
T/L
Fibroblast
Adipocytes,
Dermal and
Other Cells
BONE CARTILAGE MUSCLE MARROWTENDON/
LIGAMENT
CONNECTIVE
TISSUE
THE MESENGENIC PROCESS
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EXTRUSION-BASED PROCESSES VIII
Extruded PCL scaffolds were plasma modified with a C2H4/N2 deposition (plasma deposited ethylene: nitrogen, PdE:Ncoating) followed by H2 post treatment (PdE:N/H2 coating).
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EXTRUSION-BASED PROCESSES IX
Surface Modification Plasma Argon (Ar)
Power 100 W
Time 30 s
Frequency 0,6
Graft polymerization with Acrylic Acid (Aac)
PCL/Collagen
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EXTRUSION-BASED PROCESSES X
PCL/Collagen Incorporation of collagen
0
0,01
0,02
0,03
0,04
0,05
0,06
0,07
0,08
Day 7 Day 14
Ab
sorb
ance
PCL scaffold 0/90º PCL-col scaffold 0/90º
Human Fibroblasts (3T3)
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EXTRUSION-BASED PROCESSES XI
Scaffolds PCL/HA (75:25)
Scaffold PCL/HA Scaffold PCL
Video 2
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EXTRUSION-BASED PROCESSES XII
PCL
PCL/HA
Video 3
Scaffold PCL/Bioglass (70/30)
Video 1
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EXTRUSION-BASED PROCESSES XIII
HYDROZONE – BIOACTIVATED HIERARCHICAL HYDROGELS AS ZONAL IMPLANTS FOR ARTICULAR CARTILAGE REGENERATION. Global funding: ~12MEuros; CDRSP funding: ~1 MEuros
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EXTRUSION-BASED PROCESSES XIV
Physiological operating temperature (4ºC-37ºC) will be ensured, preserving temperature sensitive and bioactive components. Additionally, na irradiation system composed of a light source and na optical fibre attached to the extrusion heads will be implemented. This system will work as a secondary cross-linking mechanism of the extruded hydrogels, increasing the structural stability of the constructs.
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EXTRUSION-BASED PROCESSES XV
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EXTRUSION-BASED PROCESSES XVI
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EXTRUSION-BASED PROCESSES XVII
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EXTRUSION-BASED PROCESSES XVIII
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EXTRUSION-BASED PROCESSES XIX
1 - Dermal skin graft with more flexible hydrogel, entrapping fibroblasts 2 - Epidermal skin graft with stiffer hydrogel, entrapping keratinocytes
3 1
1 2
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EXTRUSION-BASED PROCESSES XX
Film
Thic
kness
in
cre
ase
(%
)
0
50
100
150
200
250
300
350
AG AGA5 AGA15 AGA25
In vitro degradation – 10 weeks in SBF solution
Degradation period (week)
0 1 2 3 4 5 6 7 8 9 10
Wei
ght
loss
(%
)
0
2
4
6
8
10
12
14
16
18
20
Film AG
Film AGA5
Film AGA15
Film AGA25
a)
Degradation period (week)
0 1 2 3 4 5 6 7 8 9 10W
ater
abso
rpti
on (
%)
0
200
400
600
800
1000
1200
1400
1600
b)
Time
Water molecules
IN
Aloe vera
OUT
Cleavable bond
Carbohydrate Polymers, In Press
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MULTI-MATERIAL LASER SINTERING FOR THE PRODUCTION OF FUNCTIONAL GRADED STRUCTURES
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MULTI-MATERIAL LASER SINTERING FOR THE PRODUCTION OF FUNCTIONAL GRADED STRUCTURES
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MULTI-MATERIAL LASER SINTERING FOR THE PRODUCTION OF FUNCTIONAL GRADED STRUCTURES
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MULTI-MATERIAL LASER SINTERING FOR THE PRODUCTION OF FUNCTIONAL GRADED STRUCTURES
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FUTURE CHALLENGES I
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FUTURE CHALLENGES II
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FUTURE CHALLENGES III
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SkelGEN - ESTABLISHMENT OF A CROSS CONTINENT CONSORTIUM FOR ENHANCING REGENERATIVE MEDICINE IN SKELETAL TISSUES.
FUTURE CHALLENGES IV
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Thank you!
The first Portuguese AM machine