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Methods in Molecular Biology 1037 Robert G. Gourdie Tereance A. Myers Editors Wound Regeneration and Repair Methods and Protocols M ETHODS IN MOLECULAR BIOLOGY™ Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK For further volumes: http://www.springer.com/series/7651 Wound Regeneration and Repair Methods and Protocols Edited by Robert G. Gourdie Virginia Tech Carilion Research Institute, Roanoke, VA, USA Tereance A. Myers School of Pharmacy, MCPHS University, Worcester, MA, USA Editors Robert G. Gourdie Tereance A. Myers Virginia Tech Carilion Research Institute School of Pharmacy, MCPHS University Roanoke, VA, USA Worcester , MA , USA ISSN 1064-3745 ISSN 1940-6029 (electronic) ISBN 978-1-62703-504-0 ISBN 978-1-62703-505-7 (eBook) DOI 10.1007/978-1-62703-505-7 Springer New York Heidelberg Dordrecht London Library of Congress Control Number: 2013941473 © Springer Science+Business Media New York 2013 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifi cally the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfi lms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. Exempted from this legal reservation are brief excerpts in connection with reviews or scholarly analysis or material supplied specifi cally for the purpose of being entered and executed on a computer system, for exclusive use by the purchaser of the work. Duplication of this publication or parts thereof is permitted only under the provisions of the Copyright Law of the Publisher’s location, in its current version, and permission for use must always be obtained from Springer. Permissions for use may be obtained through RightsLink at the Copyright Clearance Center. Violations are liable to prosecution under the respective Copyright Law. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specifi c statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. While the advice and information in this book are believed to be true and accurate at the date of publication, neither the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be made. The publisher makes no warranty, express or implied, with respect to the material contained herein. Printed on acid-free paper Humana Press is a brand of Springer Springer is part of Springer Science+Business Media (www.springer.com) Preface In this book we survey classical and cutting-edge methods for studying wound healing and regeneration. Expert scientists, bioengineers, and physicians explain useful technologies, models, techniques, and critical new areas and approaches to clinical and commercial trans- lation of research. We build on the excellent foundation provided by the 2003 publication on wound healing in the Methods in Molecular Medicine series edited by Drs. Luisa DePietro and Aime Burns. In our volume, cellular and molecular methods, genetic approaches, sur- gical techniques, clinical advances, drug discovery and delivery modalities, animal and humanized models, and new applications in the treatment of pathological wounds are cov- ered in a variety of organs and tissues. The 33 chapters are divided among sections on tissue engineering and stem cell-based approaches to wound healing, skin, corneal, and cardiovas- cular models of injury repair, nonmammalian models, and pertinent aspects of commercial- ization and intellectual property. Most chapters contain step-by-step methodological descriptions, a detailed list of materials, and an exclusive collection of personal tips or “Notes” that the authors feel critical to success in their technique. Instructive diagrams and informative color photographs also accompany the chapters. Experienced professionals in the wound healing commercial space also provide fresh insight on biotechnology startups, intellectual property, and the patent process. This compendium should enable students, trainees, established basic and clinical researchers, biomedical engineers, and entrepreneurs to effi ciently apply the trove of knowledge that has been compiled here by some of the foremost experts in wound healing today. In this way, we intend that the reader might dis- cover and translate new knowledge in the fi eld of wound repair and regeneration. Tereance A. Myers received her doctorate from Tulane University in New Orleans, LA, and did postdoctoral studies in Wound Healing Biology at the Medical University of South Carolina. Dr. Myers currently works as an Assistant Professor of Pharmaceutical Sciences at Massachusetts College of Pharmacy and Health Sciences in Worcester/Boston, MA. Robert G. Gourdie received his doctorate from the University of Canterbury in New Zealand and did postdoctoral studies in connexin biology at University College London, UK. Dr. Gourdie is currently Professor and Director of the Center for Heart and Regenerative Medicine at the Virginia Tech Carilion Research Institute and Director of Research for the Department of Emergency Medicine at the Carilion Clinic, Roanoke, Virginia. He is also Professor in the Wake Forest Virginia Tech School of Biomedical Engineering and Sciences at Virginia Tech in Blacksburg, Virginia. Roanoke, VA, USA Robert G. Gourdie Worcester, MA, USA Tereance A. Myers v Acknowledgements Drs. Gourdie and Myers wish to thank Ms. Katherine E. Degen, a graduate student at Virginia Tech in biomedical engineering for her considerable assistance in the editing of this book. vii Contents Preface. v Acknowledgements . vii Contributors. xiii PART I TISSUE ENGINEERED WOUND REPAIR MODELS 1 An In Vivo Model System for Evaluation of the Host Response to Biomaterials . 3 Brian Sicari, Neill Turner, and Stephen F. Badylak 2 Urothelial Cell Culture . 27 Yuanyuan Zhang and Anthony Atala 3 Cell-Populated Collagen Lattice Contraction Model for the Investigation of Fibroblast Collagen Interactions . 45 H. Paul Ehrlich and Kurtis E. Moyer 4 A Tissue-Engineered Corneal Wound Healing Model for the Characterization of Reepithelialization . 59 Karine Zaniolo, Patrick Carrier, Sylvain L. Guérin, François A. Auger, and Lucie Germain PART II STEM CELLS AND WOUND REPAIR 5 Adult Stem Cells in Small Animal Wound Healing Models . 81 Allison C. Nauta, Geoffrey C. Gurtner, and Michael T. Longaker 6 Novel Animal Models for Tracking the Fate and Contributions of Bone Marrow Derived Cells in Diabetic Healing . 99 Robert C. Caskey and Kenneth W. Liechty 7 Neural Repair with Pluripotent Stem Cells . 117 Máté Döbrössy and Jan Pruszak 8 Cell-Based Therapies for Myocardial Repair: Emerging Role for Bone Marrow-Derived Mesenchymal Stem Cells (MSCs) in the Treatment of the Chronically Injured Heart. 145 Jose S. Da Silva and Joshua M. Hare 9 A Model System for Primary Abdominal Closures . 165 Michael J. Yost, Mary O. Morales, Veronica Rodriguez-Rivera, Eric M. Yost, Louis Terracio, and Stephen A. Fann PART III SKIN AND CORNEAL WOUND REPAIR MODELS 10 Alternatives for Animal Wound Model Systems . 177 Phil Stephens, Matthew Caley, and Matthew Peake ix x Contents 11 Novel Methods for the Investigation of Human Hypertrophic Scarring and Other Dermal Fibrosis . 203 Dariush Honardoust, Peter Kwan, Moein Momtazi, Jie Ding, and Edward E. Tredget 12 Study of the Human Chronic Wound Tissue: Addressing Logistic Barriers and Productive Use of Laser Capture Microdissection. 233 Sashwati Roy and Chandan K. Sen 13 The Wound Watch: An Objective Staging System for Wounds in the Diabetic (db/db) Mouse Model . 245 G. Pietramaggiori, S. Scherer, and D.P. Orgill 14 Human Ex Vivo Wound Healing Model . 255 Stojadinovic Olivera and Marjana Tomic-Canic 15 Murine Models of Human Wound Healing . 265 Jerry S. Chen, Michael T. Longaker, and Geoffrey C. Gurtner PART IV CARDIOVASCULAR WOUND HEALING MODELS 16 A Corneal Scarring Model. 277 Daniel J. Gibson and Gregory S. Schultz 17 A Novel and Efficient Model of Coronary Artery Ligation in the Mouse . 299 Erhe Gao and Walter J. Koch 18 Cardiac Wound Healing Post-myocardial Infarction: A Novel Method to Target Extracellular Matrix Remodeling in the Left Ventricle. 313 Rogelio Zamilpa, Jianhua Zhang, Ying Ann Chiao, Lisandra E. de Castro Brás, Ganesh V. Halade, Yonggang Ma, Sander O. Hacker, and Merry L. Lindsey 19 Injury Models to Study Cardiac Remodeling in the Mouse: Myocardial Infarction and Ischemia–Reperfusion . 325 Daniel J. Luther, Charles K. Thodeti, and J. Gary Meszaros 20 Cryoinjury Models of the Adult and Neonatal Mouse Heart for Studies of Scarring and Regeneration. 343 Erik G. Strungs, Emily L. Ongstad, Michael P. O’Quinn, Joseph A. Palatinus, L. Jane Jourdan, and Robert G. Gourdie 21 Targeting Wnt Signaling to Improve Wound Healing After Myocardial Infarction . 355 Evangelos P. Daskalopoulos, Ben J.A. Janssen, and W. Matthijs Blankesteijn 22 Vascular Connexins in Restenosis After Balloon Injury . 381 Sandrine Morel and Brenda R. Kwak Contents xi PART V NON MAMMALIAN AND MAMMALIAN MODELS OF REGENERATIVE REPAIR 23 Gain-of-Function Assays in the Axolotl (Ambystoma mexicanum) to Identify Signaling Pathways That Induce and Regulate Limb Regeneration . 401 Jangwoo Lee, Cristian Aguilar, and