Downloaded from orbit.dtu.dk on: Sep 25, 2021 Developing 3D microstructures for tissue engineering Mohanty, Soumyaranjan Publication date: 2016 Document Version Publisher's PDF, also known as Version of record Link back to DTU Orbit Citation (APA): Mohanty, S. (2016). Developing 3D microstructures for tissue engineering. DTU Nanotech. General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. Users may download and print one copy of any publication from the public portal for the purpose of private study or research. You may not further distribute the material or use it for any profit-making activity or commercial gain You may freely distribute the URL identifying the publication in the public portal If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Developing 3D microstructures for tissue engineering Soumyaranjan Mohanty PhD Thesis March 2016 Developing 3D microstructures for tissue engineering Soumyaranjan Mohanty PhD Thesis January 2016 Developing 3D microstructures for tissue engineering By Soumyaranjan Mohanty DISSERTATION Submitted in partial fulfillment of the requirements for the degree of Doctor of Philoso- phy in Electronics, Communication and Space Science in the department of Micro-and Nanotechnology of Technological University of Denmark January 2016 ©2015 TECHNICAL UNIVERSITY OF DENMARK. All rights reserved. Supervised by: Associate Professor Anders Wolff Professor Jenny Emnéus Associate Professor Martin Dufva Contents Summary ............................................................................................................................ 1 Resumé .............................................................................................................................. 3 Acknowledgments ............................................................................................................. 5 List of abbreviations .......................................................................................................... 6 List of Publications and Submitted Manuscripts ............................................................... 8 1. CHAPTER 1. Introduction: Scope of the Thesis ....................................................... 11 1.1 Various extracorporeal liver support systems .................................................. 11 1.2 Sources of hepatocytes for large scale EBALs ................................................ 13 1.3 EU project - NanoBio4Trans ........................................................................... 15 1.4 Motivation and Objective................................................................................. 16 1.5 Thesis Outline .................................................................................................. 18 1.6 References ........................................................................................................ 20 2. CHAPTER 2. State of the art: Scaffold Design Parameter and Fabrication ............. 24 2.1 Introduction ...................................................................................................... 24 2.2 Scaffold fabrication methods ........................................................................... 25 2.2.1 Random Porous Scaffold Fabrication: Using Conventional Methods . 26 2.2.2 Structured porous scaffold fabrication: using solid free-form fabrication / rapid prototyping (RP) ..................................................... 29 2.2.3 Next generation smart scaffolds: integration of multiple fabrication methods ................................................................................................ 38 2.3 Conclusions ...................................................................................................... 46 2.4 References ........................................................................................................ 46 3. CHAPTER 3. Fabrication of Scalable and Structured Tissue Engineering Scaffolds Using Water Dissolvable Sacrificial 3D Printed Moulds .......................................... 55 3.1 Introduction ...................................................................................................... 55 3.2 Materials and Methods ..................................................................................... 57 3.3 Results .............................................................................................................. 64 3.4 Discussion ........................................................................................................ 72 3.5 Conclusions ...................................................................................................... 75 3.6 References ........................................................................................................ 75 4. CHAPTER 4. Fabrication of scalable tissue engineering scaffolds with dual-pore microarchitecture by combining 3D printing and particle leaching .......................... 83 4.1 Introduction ...................................................................................................... 83 4.2 Materials and method ....................................................................................... 85 4.2.1 Mould fabrication ................................................................................. 85 4.2.2 Characterizations of scaffolds .............................................................. 87 4.2.3 Cell culture and assays in scaffolds ..................................................... 88 4.3 Results and discussion ..................................................................................... 91 4.3.1 Scaffold fabrication .............................................................................. 91 4.3.2 Scaffold characterization...................................................................... 95 4.3.3 Cell proliferation and viability ............................................................. 98 4.4 Conclusions .................................................................................................... 104 4.5 References ...................................................................................................... 105 5. CHAPTER 5. A 3D printed silicone-hydrogel scaffold with enhanced physicochemical properties ..................................................................................... 114 5.1 Introduction .................................................................................................... 114 5.2 Experimental Section ..................................................................................... 116 5.2.1 Materials ............................................................................................ 116 5.2.2 Fabrication of 3D elastomer scaffolds ............................................... 117 5.2.3 Fabrication of IPNs ............................................................................ 117 5.2.4 Water uptake kinetics ......................................................................... 118 5.2.5 Surface characterization ..................................................................... 118 5.2.6 Mechanical properties ........................................................................ 119 5.2.7 In-vitro cell studies ............................................................................ 119 5.2.8 Doxycycline drug release studies ....................................................... 121 5.2.9 Induction of gene expression of Hela Tet-On cells through doxycycline release from IPNs ............................................................................... 121 5.2.10 Statistical analysis .............................................................................. 122 5.3 Results and Discussion .................................................................................. 123 5.3.1 Fabrication of 3D IPN scaffolds ........................................................ 123 5.3.2 Physicochemical characteristics and mechanical properties of 2D IPNs.124 5.3.2 In-vitro cell studies on pHEMA-co-PEGMEA 2D and 3D IPN materials ............................................................................................. 126 5.3.3 DOX release studies from IPNs ......................................................... 131 5.4 Conclusions .................................................................................................... 133 5.5 Associated Content ........................................................................................ 134 5.6 References ...................................................................................................... 135 6. CHAPTER 6: Application of 3D porous scaffolds in the NanoBio4Trans project for perfusion bioreactor array with integrated sensors and differentiation of iPSC into hepatocytes. ............................................................................................................. 141 6.1 Perfusion bioreactor array for tissue engineering applications ...................... 141 6.2 Impedance based sensor integration and monitoring of cell growth in BAL support system ............................................................................................... 143 6.3 Assessment