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

277 Pages·7.021 MB·English

by  Vrana N.E.

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
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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.

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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.
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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.

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Awards

1) Finalist in Sir Bernard Crossland first year research awards (2007)
2) Finalist in TERMIS SYIS- Project proposal competition with two projects (2008)

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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
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