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ABSTRACT HE, TING. Tissue Engineering 3D Textiles Scaffolds ABSTRACT HE, TING. Tissue Engineering 3D Textiles Scaffolds for Multiple Junction Tissue Regeneration. (Under the direction of Dr. Martin W. King and Dr. Robert Dennis). The concept of using a heterogeneous or multiphase scaffold containing a structural and functional gradient is essential in the regeneration of interfacial tissue engineering such as a muscle-tendon junction (MTJ). The primary goal of this study was to use appropriate textile technologies to engineer an integrated and contiguous multiphase scaffold that can serve for MTJ regeneration. The latest warp knitting technology was used to develop a multiphase spacer fabric scaffold with an appropriate gradient of structure and properties that mimic the natural MTJ junction tissue. Four knitting methods were developed to fabricate the multiple phase spacer fabric scaffolds. The spacer fabric scaffolds were evaluated in terms of their physical and mechanical properties. Murine myoblasts and fibroblasts were co-cultured on the preferred MTJ spacer fabric scaffolds under both static and cyclic stretching conditions that represented dynamic mechanical stimulation for the natural MTJ tissue. The biological properties of the co-cultured spacer fabric scaffolds were monitored by scanning electronic microscopy (SEM), routine histology and quantitative polymerase chain reaction (qPCR). A novel approach using laser scanning confocal microscopy (LSCM) together with fluorescent cell labelling was developed to create 3D images of the cell morphology for the regenerated muscle-tendon junction tissue on and within the spacer fabric scaffold. A compact bioreactor system has been developed to provide cyclic stretching stimulation during dynamic muscle tendon junction co-culture. This study also briefly investigated the isolation and culture of porcine primary muscle and tendon cells, which points to an alternative primary cell source for future studies using a pig model. This research is innovative in designing and engineering textile scaffolds and a dynamic cyclic stretching bioreactor, as well as establishing a systematic approach for the fabrication and evaluation of MTJ tissue engineering constructs. © Copyright 2016 by Ting He All Rights Reserved Tissue Engineering 3D Textiles Scaffolds for Multiple Junction Tissue Regeneration by Ting He A dissertation submitted to the Graduate Faculty of North Carolina State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy Fiber and Polymer Science Raleigh, North Carolina 2016 APPROVED BY: _______________________________ _______________________________ Dr. Martin W. King Dr. Robert Dennis Co-chair of Advisory Committee Co-chair of Advisory Committee _______________________________ _______________________________ Dr. Warren Jasper Dr. Stephen Michielsen ii DEDICATION 老爸, 我是何博士啦 ~你看到了吗？ 爱你~ ^_^ iii BIOGRAPHY Ting He was born January 19, 1987 in Xuzhou, Jiangsu, China. She attended Xuzhou Senior High School in Xuzhou. During her undergraduate program in the College of Textiles at Donghua University in Shanghai, China, she was selected to attend the 3+X exchange program with North Carolina State University (NCSU) in 2009. Ting received her Bachelor of Science in Textile Engineering from Donghua University in 2010. Then she was admitted by the graduate program in the College of Textiles at NCSU where she undertook research on biomedical textiles under the supervision of Prof. Martin W. King. After receiving her Master of Science in Textile Engineering, she continued her research in biomedical textiles by enrolling in the doctoral program in Polymer and Fiber Science in the College of Textile, NCSU. iv ACKNOWLEDGMENTS As I wrap up my scientific research study and begin to give thought to my life and work over the past a few years, many names and faces flash into my head, reminding me of the generous support, encouragement and guidance I have been granted along this journey of research. A special thank you is extended to Dr. Martin W. King, Co-chair of my Advisory Committee. His knowledge and expertise lead me to the frontier of biomedical textiles (BMT). His patience and consideration paved the rugged road during my journey. He has taken care of every student in our BMT research group like a family. He has been so respectful, supportive and caring and has shared all the joys and concerns in the life and research of our BMT group family. I am grateful for the support and help from Dr. Robert Dennis, Co-chair of my Advisory Committee. With his extensive experience in cell biology and mechanical engineering, my research study has been expanded and brought to life. My sincerely appreciation is extended to my dissertation committee Dr. Warren Jasper and Dr. Stephen Michielsen, as well as the Graduate School Representative, Dr. Hsiao-Ying Huang. The interest they displayed in my research and the time they spent on evaluating my proposal and dissertation has motivated and encouraged me through many challenges. A great "Thank you" to the team at Wake Forest Institute of Regenerative Medicine, particularly Dr. Sang Jin Lee, Dr. Young Min Ju and Denethia Green. I have valued their participation and input and have appreciated their expertise. Many thanks to all the group members in Dr. Martin W. King’s Biomedical Textiles (BMT) Research Group for their company, encouragement and support. v Finally, my deepest appreciation is extended to my husband and my mother. They accompanied me through the long journey and stayed by my side offering warmest support. Their love and trust supported me through the difficult times. My gratitude for them cannot be expressed in any language. vi TABLE OF CONTENTS LIST OF FIGURES ..................................................................................................................................... ix LIST OF TABLES ...................................................................................................................................... xv CHAPTER 1 ................................................................................................................................................. 1 INTRODUCTION ........................................................................................................................................ 1 CHAPTER 2 ................................................................................................................................................. 6 LITERATURE REVIEW ............................................................................................................................. 6 Overview ....................................................................................................................................... 6 Interfacial Tissue Engineering and Multiple Tissue Junctions ..................................................... 7 Interfacial Tissue Engineering (ITE) .................................................................................... 7 Muscle-tendon junctions (MTJ) ............................................................................................ 9 Textile Scaffolds for Interface Tissue Engineering............................................................. 17 Scaffold Fabrication Technologies ..................................................................................... 33 Textile interface tissue engineering (ITE) scaffolds ........................................................... 39 Warp Knitted Spacer Fabric Scaffolds ........................................................................................ 46 Biotextile Fabrication Technologies ................................................................................... 46 Systematic Approach to Tissue Engineering a Muscle Tendon Junction (MTJ) ........................ 56 Developmental Strategy and Methods ................................................................................ 56 Summary ..................................................................................................................................... 65 CHAPTER 3 ............................................................................................................................................... 67 DEVELOPMENT OF MUSCLE-TENDON JUNCTION SPACER FABRIC SCAFFOLD ..................... 67 Introduction ................................................................................................................................. 67 Design of Multiple Phase Spacer Fabric ..................................................................................... 68 Muscle-tendon Junction (MTJ) Tissue Model .................................................................... 68 Development of Non-degradable Surrogate Prototype ............................................................... 70 Materials ............................................................................................................................. 70 Knitting Method .................................................................................................................. 71 Fabrication .......................................................................................................................... 76 Evaluation of Non-degradable Polymer ...................................................................................... 77 Methods ............................................................................................................................... 77 Results and Discussion.......................................................................................................
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