Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free...

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Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing

Transcript of Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free...

Page 1: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Yong HuangDepartment of Mechanical

Engineering

Clemson University, Clemson, SC

Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing

Page 2: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Table of Contents Introduction Background Objective Research Tasks

Tube Inkjetting Tube Laser Printing

Conclusions Future Work

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Page 3: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Table of Contents Introduction Background Objective Research Tasks

Tube Inkjetting Tube Laser Printing

Conclusions Future Work

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Page 4: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Background Inkjet-based

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[Boland, 2007]

Inkjetting of CaCl2 into 2% alginic acid to make a branched structure

[Nakamura, 2008]

Inkjetting 0.8% sodium alginate into

2% CaCl2 to make a tube with the help of gravity

Page 5: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

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[Skardal, 2010]

Printing of agarose and 3T3 cell-containing PEGTA filaments to make a tube

[Norotte, 2009]

Printing of agarose and smooth muscle cell (SMC) cylinders to make a tube

Background Filament-based

Page 6: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Background Laser-based

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Laser printing of alternating layers of fibroblasts (green) and keratinocytes (red) to form a grid structure (Scale bar = 500 µm)

[Koch, 2012]

Page 7: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Challenges

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How to scale-up scaffold-free bioprint complex structures such as overhang structures?

How to evaluate and mitigate the post-printing cell damage?

Page 8: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Table of Contents Introduction Background Objective Research Tasks

Tube Inkjetting Tube Laser Printing

Conclusions Future Work

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Page 9: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Objective

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Needs in cellular tube bioprintingVascularization as a main technological barrier for building

3D organs

Tube printing - a logical initial step towards vascularization

Objective

To scaffold-free fabricate viable cellular tubes using inkjet and laser printing

Page 10: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Table of Contents Introduction Background Objective Research Tasks

Tube Inkjetting Tube Laser Printing

Conclusions Future Work

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Page 11: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Table of Contents Introduction Background Objective Research Tasks

Tube Inkjetting Tube Laser Printing

Conclusions Future Work

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Page 12: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

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Liquid levelFirst layerSecond layer

3T3 cell suspension

Tube Inkjetting Experimental setup

Page 13: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Tube Inkjetting - Zig-zag tube

Fabrication process: bottom part, overhang part and top part

Wall thickness: 150~200 µm

Tube diameter: 3 mm and length: 10 mm

Zigzag tube fabrication

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Page 14: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Tube Inkjetting - Cell viability test

(a) Printed tube, (b) tube surface and (c) cell viability test after liquefying

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(a)

Cells

(b)

Live Cell

Dead Cell

After liquefying

(c)

Page 15: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Cell viability immediately after printing: > 85% Cell viability after 3 days: > 80% Cell viability comparable to that of the control

Tube Inkjetting - Cell viability

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0 24 48 7270

75

80

85

90

95

Cel

l via

bilit

y %

Post-printingControl

(Hours)

Page 16: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Tube Inkjetting - Other structures

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Pyramid Cone

Page 17: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Table of Contents Introduction Background Objective Research Tasks

Tube Inkjetting Tube Laser Printing

Conclusions Future Work

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Page 18: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Tube laser printing - Experimental setup

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ArF laser

Pulsed laser beam

Ribbon

Forming jet/droplet

Quartz support

Coating

Container

Optical table

Laser-induced vapor/plasma

pocket

3-axis stages

Moving platform

Page 19: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Tube laser printing - Experimental setup

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Direct-writing height = 1 mm

ArF laser

Pulsed laser beam

Ribbon

Forming jet/droplet

Quartz support

Coating

Container

Optical table

Laser-induced vapor/plasma

pocket

3-axis stages

Moving platform

Page 20: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Tube laser printing - results

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Top viewSide view

Φ=3 mm

Page 21: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Table of Contents Introduction Background Objective Research Tasks

Tube Inkjetting Tube Laser Printing

Conclusions Future Work

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Page 22: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Overhang structures can be scaffold-free fabricated

Viable cellular tubes (3T3) can be printed

ConclusionsInkjetting

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Tubes can be scaffold-free fabricated using LIFT-based laser printing technology

Highly viscous materials (alginate) can be laser printed into well-defined tubular structures

Laser printing

Page 23: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Table of Contents Introduction Background Objective Research Tasks

Tube Inkjetting Tube Laser Printing

Conclusions Future Work

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Page 24: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Laser printing of 3D cellular tubes

Printing of adipose-derived stem cell tubes

Future WorkFabrication

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Apoptosis/necrosis test, cell proliferation capacity, and phenotype variation

Post-printing fusion of tubular tissue

Mechanical property measurement of fabricated tubes

Post-printing evaluation

Page 25: Yong Huang Department of Mechanical Engineering Clemson University, Clemson, SC Scaffold-free Alginate Tube Fabrication using Inkjet and Laser printing.

Thanks

and questions?

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