3d druck und health techcpd) cell-compatible printing process bioinktm ……... bioinktm...

28.08.2014 Health Tech Cluster Switzerland: 3D Druck und Health Tech

Additive Verfahren mit biologischen Materialien

Dr. Markus Rimann Zürcher Hochschule für Angewandte Wissenschaften (ZHAW) Fachgruppe Tissue Engineering

3D Druck und Health Tech


Background: 3D Cell Culture

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Why should we cultivate cells in three-dimensions (3D)? • Humans are 3D

Advantages of 3D cell culture • Cell behavior between 2D and 3D is completely different (Proliferation, differentiation, metabolism) • Different metabolic activity of 2D versus 3D, e.g. higher drug resistance of 3D • Cell communication between cells is decreased in 2D, but crucial.

Drawbacks of current 3D cell culture technologies • Limited control of cell distribution within a 3D system • Integration of blood vessels difficult (to control) (Tissue size limited, O2- and nutrient limitation)

=> Artificial organs difficult

Application in regenerative medicine and drug testing • Provides reliable data • Reduces animal experiments • Accelerates cost-intensive process of drug development

2D cell culture

3D cell culture

Organs on demand Regenerative medicine

Drug testing

High organ structure

complexity

Schematc representation of kidney structure (ww.arizonatransplantassociates .com)

Schematic representation of skin structure (www.intechopen.com)

28.08.2014 Health Tech Cluster Switzerland: 3D Druck und Health Tech 3

Background: Organs on demand

Principle: Layer-by-layer deposition of biological material to produce 3D organotypic in vitro models

Flexibility in 3D shape

Spatial control of cells, matrix and bioactive molecules (growth factors etc.)

Standardized, automated and reproducible process

High-throughput screening

Integration of quality control

Guillotin and Guillemot, 2011 3DDiscovery® bioprinter, regenHU (www.regenhu.com)

Bioprinting

Is the bioprinting technology the solution?


Standardization of a printing process for the manufacture of soft tissue models

Approved printing device

Cell-compatible bioink

Choice of a soft tissue model as proof of concept

Keratinocytes seeding

Skin

. . . . . . . . . . . . . . . . . . . . .

Fibroblasts in collagen I gel

Contraction

Keratinocytes on top of dermis

Air-lift

Keratinocytes differentiation

Contracted collagen I gel

3DDiscovery®

User-friendly software

Sterile printing conditions

Printing process in standard cell culture devices

3DDiscovery® printer, regenHU

BioInkTM

BioInkTM

cartridge, regenHU


ZHAW research


ZHAW research

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Bioprinter

• Sterile printing environment • Cell-compatible printheads • User-friendly software (CAD-program) • Flexible system to cope with fast development of this

technology

Bioprinting technologies (Malda et al., 2013, Adv. Mat.)


ZHAW research

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Bioink

• Printable matrix to arrange cells in 3D (universal bioink) • Cell-compatible, cell adhesion, cell migration • Stiffness adjustable (bone, soft tissue) • Fast polymerization (different modes) • Ready-to-use commercial product

Printed structures with bioink (Melchels et al., 2014, J. Mater. Chem. B.)

BioInk™

Dermis equivalent scheme

Human primary fibroblasts printed in BioInkTM

dermis equivalents. Models stained with MTT after 2 days of culture show cell viability and a clear printing pattern

The polymerization process of the matrix at 365nm ± 10nm doesn’t cause DNA damages. ELISA analysis for the detection of cyclobutane pyrimidine dimers (CPD)

Cell-compatible printing process

BioInkTM

BioInkTM ……...

Fibroblasts

ZHAW research



International research

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Complex tissue structures with “blood vessels” Kolesky, Truby, Gladman, Busbee, Homan, Lewis Wyss Institute for Biologically Inspired Engineering, Harvard University, USA (Adv. Mat., 2014)

Tissue production by extrusion technology (robotic dispensing, slide 6). Left: Principle of a printed construct (ECM: extracellular matrix), right top: Scheme of the printed structure, right bottom: Printed construct with differently colored cell types.

Different bioinks are printed together Different cell types are precisely deposited within a 3D structure Blood vessel printing is feasible (not in one step) Larger tissue constructs are printable because of vasculature integration



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Printed liver

Liver complexity (http://www.cosmiq.de/qa/show/2802267/hat-der-mensch-eine-leber-oder-zwei/)

Printed „liver“ harboring three different cell types. Organovo, San Diego, CA 92121, USA

Three different cell types were printed in the same construct Liver-specific functions were reported Liver complexity not achieved



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Printed kidney

Kidney complexity (http://www.jameda.de/gesundheits-lexikon/niere/)

Printed „kidney“. Atala, Wake Forest Institute for Regenerative Medicine, Winston Salem, NC 27157, USA,

Printed kidney shape harboring cells Kidney complexity not achieved Kidney functionality not achieved


Summary

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• Bioprinting has a huge potential in substance testing and regenerative medicine

• Technology is still in its infancy

Printing of simple structures (resolution?)

• Lack of standardization and reproducibility

Research groups are fabricating their own bioprinters Research groups are synthesizing their own bioinks

• Few commercially-available bioprinters

Technology at high level

• Few commercially-available bioinks

Cell-compatibility is important and difficult to achieve Thorough material characterization is necessary (stiffness, viscosity, degradability)

Bioink properties (Malda et al., 2013, Adv. Mat.)


Acknowledgements

Prof. Dr. Ursula Graf-Hausner Dr. Epifania Bono Helene Annaheim Matthias Bleisch Tissue Engineering Team

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Marc Thurner Michael Kuster Andreas Scheidegger

Danke für Ihre Aufmerksamkeit

3d druck und health techcpd) cell-compatible printing process bioinktm ……... bioinktm...

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