MECHANICAL CHARACTERIZATION OF BIOPOLYMER AND THE EFFECTS OF MECHANICAL PROPERTIES OF EXTRACELLULAR MATRIX ON CELL BEHAVIORS WONG LONG HUI NATIONAL UNIVERSITY OF SINGAPORE 2013 MECHANICAL CHARACTERIZATION OF BIOPOLYMER AND THE EFFECTS OF MECHANICAL PROPERTIES OF EXTRACELLULAR MATRIX ON CELL BEHAVIORS WONG LONG HUI (B.Eng.(Hons.), NTU) A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN CHEMICAL AND PHARMACEUTICAL ENGINEERING SINGAPORE-MIT ALLIANCE NATIONAL UNIVERSITY OF SINGAPORE 2013 Declaration I hereby declare that this thesis is my original work and it has been written by me in its entirety I have duly acknowledged all the sources of information which have been used in the thesis This thesis has also not been submitted for any degree in any university previously _ Wong Long Hui 2nd October 2014 i Acknowledgments The work presented herein could not have been completed without the guidance and support of numerous individuals and collaborators The brilliance, humor and encouragement of these people have not only made possible the existence of this thesis but also crafted my graduate school experience into an enriching and memorable one First and foremost, I would like to thank Professor Too Heng-Phon Through his infectious enthusiasm, never-ending reminder of “lag-time” theory and superhuman work ethic, I have learned both the complexity of science and profundity of life Prof Too’s lessons extended far beyond the laboratory – from politics and current issues to virtues of integrity, honest work and humility Especially striking to heart is his emphasis that hardwork is a part of real and successful life From the adhoc “latte” time to the regular Friday snub time, Prof Too’s clarity of thoughts, deep insights and multiple perspectives to any issues have never failed to astound me I thank him for teaching me how to optimize and be obsessed with my life I hope that one day I will be able to influence another’s life in the same way A substantial part of this thesis was written with the theoretical insights of Prof Raj Rajagopalan as well I thank Prof Raj for acquainting me with the importance of clarity of thoughts in effective communication, through his particular emphasis on eloquence in writing It has always been incredibly fun and insightful to chat with Prof Raj because of his exceptional sense of humor and great wisdom I thank him for taking time to broaden my horizon to the amazing scientific world through his extensive experience and wealth of knowledge I am also indebted to Nicholas Agung Kurniawan for taking me under his wing during the ii initial years of my postgraduate school I would never forget the number of hours spent in the rheometer lab with Nicholas; I consulted with him on one too many mechanical engineering theories and perspectives of both life issues and scientific works I truly cannot have asked for a more talented and embracing mentor The members of Professor Too’s group have provided an extraordinarily supportive and intellectually-stimulating environment, which has expedited my immersion into the biology field and provided me with alternative insights to my engineering viewpoints I am especially thankful to Chen Xixian for her unfailing moral support and countless exchanges ever since our first encounter in the athenaeum I also thank Sarah Ho Yoon Khei for tutoring me in the initial lab culture techniques and battering with me over countless scientific issues from both the biological and engineering perspectives Finally, I would like to express my gratitude and affection to my parents, my grandma, my two brothers and my boyfriend They have provided me with unconditional love and unstinted support during these years I would most like to thank my parents for believing in me and providing a warm and caring home to return to every night This thesis is dedicated to my parents Financial support for this work provided by the Singapore – Massachusetts Institute of Technology Alliance is gratefully acknowledged iii Publications Published works: Wong, L H., Kurniawan, N A., Too, H -P., and Rajagopalan, R Spatially resolved microrheology of heterogeneous biopolymer hydrogels using covalently bound microspheres Biomechanics and Modeling in Mechanobiology (Chapter 3) Kurniawan, N A., Wong, L H, and Rajagopalan, R (2012) Early Stiffening and Softening of Collagen: Interplay of Deformation Mechanisms in Biopolymer Networks Biomacromolecules, 13, 691-698 (Chapter 2) Submitted manuscript: Wong, L H., Chung, C W., Kurniawan, N A., Too, H –P, Buist, M L., and Rajagopalan, R Hierarchical structure of deformation mechanisms in increasing shear straining of collagen network (Chapter 2) Wong, L H., Ho, Y K., and Too, H -P Acquired temozolomide resistance induces an aggressive, migrative and invasive phenotype in U251MG human cell line (Chapter 4) iv Manuscripts in preparation: Wong, L H., Kurniawan, N A., Too, H -P., and Rajagopalan, R Correlating pore size heterogeneity to micromechanics derived using cross-linked microspheres (Chapter 3) Ho, Y K., Wong, L H., Rajagopalan, R., and Too, H -P TrafEnTM- A novel reagent to achieve synergism of polymer based transfection and chemosensitization for osteogenic differentiation of mesenchymal stem cells in three dimensional cultures (in preparation) v Conferences Wong, L H., Chung, C W., Kurniawan, N A., Too, H –P, Buist, M L., and Rajagopalan, R Modelling viscoelastic properties of collagen networks under dynamic shear loading The 15 th International Conference on Biomedical Engineering (ICBME 2013), Singapore, 4th – 7th Dec 2013 Wong, L H., Ho Y K., and Too, H –P Microenvironment cues induce invasiveness in acquired Temozolomide resistance glioma cells The European Cancer Congress (ECCO 2013), Amsterdam, The Netherlands, 27th Sept – 1st Oct 2013 Wong, L H., Ho Y K., and Too, H –P Microenvironment cues induce invasiveness in acquired Temozolomide resistance glioma cells Biomedical Engineering Singapore th Student Symposium (BES7SM), Singapore, 18th May 2013 Wong, L H., Hatton, T A., and Too, H –P Spatially-resolved microrheology with covalently-bound microspheres, Singapore – MIT Alliance Symposium 2013, Singapore, 14th – 15th Jan 2013 Wong, L H., Too, H –P, Rajagopalan, R Enhancing particle-tracking microrheology technique with covalently-bound probes, Biomedical Engineering Singapore 6th Student Symposium (BES6SM), Singapore, 19th May 2012 vi Wong, L H., Too, H –P, and Rajagopalan, R Cellular motility and micromechanics of extracellular matrices, Singapore – MIT Alliance Symposium 2011, Singapore, 10th Jan 2011 Wong, L H., Kurniawan, N A., Rajagopalan, R., and Too, H –P Spatially-resolved microrheology of heterogeneous biopolymer hydrogels with covalently-bound microspheres, 5th East Asian Pacific Student Workshop on Nano-Biomedical Engineering, Singapore, 12th – 14th Dec 2011 vii List of Awards Travel Award Singapore – MIT Alliance The 15th International Conference on Biomedical 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