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Use of Poly Vinyl Alcohol (PVA) Cryogelation for Tissue Engineering: Composites, Scaffold Formation and Cell Encapsulation PDF

277 Pages·2016·7.02 MB·English
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Use of Poly Vinyl Alcohol (PVA) Cryogelation for Tissue Engineering: Composites, Scaffold Formation and Cell Encapsulation Nihal Engin Vrana (M. Sc. BTech) PhD 2009 Use of PVA Cryogelation for Tissue Engineering: Composites, Scaffold Formation and Cell Encapsulation By Nihal Engin Vrana (M.Sc. BTech) Thesis presented to Dublin City University in fulfilment of the requirements for the degree of Doctor of Philosophy Supervisors: Dr. Garrett B. McGuinness Professor Paul A. Cahill School of Mechanical and Manufacturing Engineering, Dublin City University, Ireland September 2009 Declaration I hereby certify that this material, which I now submit for assessment on the programme of study leading to the award of Doctor of Philosophy (Ph.D.) is entirely my own work, that I have exercised reasonable care to ensure that the work is original, and does not to the best of my knowledge breach any law of copyright, and has not been taken from the work of others save and to the extent that such work has been cited and acknowledged within the text of my work. Signed: ____________ (Nihal Engin Vrana) ID No.: ___________ Date: _______ I Acknowledgements First and foremost, I would like to thank my supervisors Dr.Garrett McGuinness and Prof. Dr. Paul Cahill for their help, advice and support over the last 3 years. I also want to acknowledge the help of Dr. Lisa Looney, Dr. Joseph Stokes, Dr. Dermot Brabazon and Dr. Triona Lally and Dr. Sumsun Naher in Mechanical and Manufacturing engineering department. Also, thanks to Dr Tony O’Grady and Prof. Elaine Kay for their help with the histology experiments. My technicians Jim Barry and Michael May were also very helpful when I needed parts from the workshop. My fellow PhD students, Diana Garcia Alonso Garcia, Szilvia Eosoly, Tamas Szucs, Irina Pascu, Marcin Lipowiecki, Alberto Colombo, Vittoria Flammini and especially Yurong Liu; thank you for your support and friendship. I hope I have helped you as much as you helped me. A big thanks to my colleagues in VHRC, Shaunta Guha, Mishan Britto, Paul Fitzpatrick, Tony Walsh, Anthony Guigan, Andrew, Maria Kileen and also Dr. Phil Cummins, Dr. Ronan Murphy and Dr. Sinead Loughran for their help with the instruments. I would like to thank my girlfriend, Alix Stanciu, for her support over the last year; especially for the last 3 months when she needed to deal with a grumpier version of me. It is like putting on weight; you never realize how boring you get and I really appreciate her patience and understanding. A nod of appreciation to my band of brothers who never ceases to support me no matter where they are. They have II always kept this world fresh for me with their strange projects and aspirations; it is wonderful to know that someone is rooting for you and it is equally wonderful to root for someone when things are not very bright for you. Finally, I would also want to thank my triumvirate of mothers, Fusun Sonar, Sevim Sonar and Funda Tafulcan, for their unwavering belief in me that always has made me to live up to it. A huge thanks to my late grandfather, Nihal Sonar too, for infusing with the idea of perseverance that I tap into more than I have ever imagined. There is really a way when there is a will. This research has been supported by a Marie Curie Early Stage Research Training Fellowship of the European Community's Sixth Framework Programme under contract number MEST-CT-2005-020621. III Publications and Presentations Publications 1) N.E. Vrana, Y. Liu, P.A. Cahill, G.B. McGuinness. Characterization of PVA (Polyvinyl Alcohol)/Chitosan hydrogels as Vascular Tissue Engineering Scaffolds Macromolecular Symposia 2008: 269: 106-110. 2) Y. Liu, N. E. Vrana, P.A. Cahill, G.B. McGuinness. Physically crosslinked combined cryogels of PVA and natural macromolecules: physical and endothelial cell culture characteristics Journal of Biomedical Materials Research Part B: Applied Biomaterials 2009: 90B (2): 492-502. 3) N.E. Vrana, A. O’Grady, E. Kay, P.A. Cahill, G.B. McGuinness. Cell Encapsulation within PVA-based Hydrogels via Freeze-Thawing: A One-step Scaffold Formation and Cell Storage Technique, Journal of Tissue Engineering and Regenerative Medicine 2009, in press. 4) N.E. Vrana, P.A. Cahill, G.B. McGuinness. Endothelialisation of PVA/Gelatin Hydogel Scaffolds: Effect of Turbulent Flow Induced Shear Stress, in preparation. 5) N.E. Vrana, P.A. Cahill, G.B. McGuinness. Characterisation of Vascular Smooth Muscle Cell Encapsulated PVA/Gelatin Cryogels and Their Endothelialisation, in preparation. IV Presentations 1) N.E. Vrana, P.A. Cahill, G.B. McGuinness 2007. Characterization of PVA/Chitosan Hydrogels as Vascular Tissue Engineering Scaffolds, 4th European Symposium on Biopolymers, Kusadasi, Turkey, October 2007. 2) N.E. Vrana, P.A. Cahill, G.B. McGuinness 2008. Effect of Composition of PVA- based Hydrogels on vascular cell behaviour, 14th Bioengineering Ireland Conference, Sligo, Ireland, January 2008. 3) N.E. Vrana, Paul Cahill, Garrett McGuinness 2008. Investigation of PVA based Hydrogels for Vascular Tissue Engineering, 11th Sir Bernard Crossland Symposium, Limerick, Ireland, March 2008. 4) N.E. Vrana, P. Cahill, G. McGuinness 2008. Behaviour of encapsulated vascular smooth muscle cells and endothelial cells on PVA/Gelatin cryogels, TERMIS-EU 2008, Porto, Portugal, June 2008. 5) N.E. Vrana, Y. Liu, P. Zorlutuna 2008. Effect of Blood-like nano/micro bioactive molecule loaded particle suspension on the behaviour of PVA-based hydrogels under pulsatile flow conditions, TERMIS-EU 2008, Porto, Portugal, June 2008. 6) N.E. Vrana, Y. Liu, P. Zorlutuna 2008. Controlling the vascular smooth muscle cell phenotype in photopolymerisable electrospun nanofibrous mats by microenvironmental mechanical changes, TERMIS-EU 2008, Porto, Portugal, June 2008. V 7) N.E. Vrana, P. Cahill, G. McGuinness 2009. Cell Encapsulation within PVA-based Hydrogels via Cryogelation:A One-Step Scaffold Formation and Storage Technique. 15th Bioengineering Ireland Conference, Limerick, Ireland, January 2009. 8) N.E. Vrana, P. Cahill, G. McGuinness 2009. Vascular Smooth Muscle Cell Encapsulation via PVA Cryogelation. TERMIS-World, Seoul, South Korea, August 2009. 9) N.E. Vrana, P. Cahill, G. McGuinness 2009. Endothelialisation of PVA-based Vascular Tissue Engineering Scaffolds under Turbulent Flow. ESB 2009, Lausanne, Switzerland,September 2009. VI Awards 1) Finalist in Sir Bernard Crossland first year research awards (2007) 2) Finalist in TERMIS SYIS- Project proposal competition with two projects (2008) VII Table of Contents Declaration ..................................................................................................................... I Acknowledgements ....................................................................................................... II Publications and Presentations .................................................................................... IV Awards ......................................................................................................................... VII List of Abbreviations ....................................................................................................... XIV Nomenclature ............................................................................................................. XVI List of Figures ............................................................................................................ XVIII List of tables.............................................................................................................. XXIV Glossary ......................................................................................................................... XXV Abstract ........................................................................................................................ XXVI CHAPTER 1 ......................................................................................................................... 1 1 Introduction .................................................................................................................... 1 1.1Objectives of the Research Project .......................................................................... 4 CHAPTER 2 ....................................................................................................................... 10 2 Literature Review ......................................................................................................... 10 2.1 Cardiovascular Diseases and Current Remedies ................................................... 10 2.2 Structure and Composition of Vascular Tissue ...................................................... 17 2.2.1 Endothelial Cells ............................................................................................. 19 2.2.2 Smooth Muscle Cells ...................................................................................... 23 2.3 Vascular Tissue Engineering .................................................................................. 27 2.3.1 Aims and Requirements of Vascular Tissue Engineering ............................... 27 VIII

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Thesis presented to Dublin City University in fulfilment of the requirements for the degree of Doctor of Philosophy - School of Mechanical and Manufacturing Engineering, Dublin City University, Ireland – 2009. – 277 p.PVA cryogelation is a physical hydrogel formation method, which yields cryogel
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