EFFECT OF NORIAN® CRS® FAST SET™ PUTTY ON IN VIVO OSSEOUS REPAIR USING A CRITICAL SIZE DEFECT IN THE RATTUS NORVEGICUS CALVARIUM By Richard L. Williams, D.M.D., M.Ed. Major, US Army Dental Corps Submitted to the Faculty of the School of Graduate Studies of the Medical College of Georgia in partial fulfillment of the Requirements for the Degree of Master of Science FEBRUARY 2005 EFFECT OF NORIAN® CRS® FAST SET™ PUTTY ON IN VIVO OSSEOUS REPAIR USING A CRITICAL SIZE DEFECT IN THE RATTUS NORVEGICUS CALVARIUM This thesis is submitted by Richard L. Williams and has been examined and approved by an appointed committee of the faculty of the School of Graduate Studies of the Medical College of Georgia. The signatures, which appear below, verify the fact that all required changes have been incorporated and that the thesis has received final approval with reference to content, form and accuracy of presentation. This thesis is therefore in partial fulfillment of the requirements for the degree of Master of Science. ' ate_... TABLE OF CONTENTS ACKNOWLEDGEMENTS ................................................................. iv LIST OF FIGURES ......................................................................... vii LIST OF TABLES ........................................................................... ix INTRODUCTION A. Statement of the Problem ...................................... 1 B. Review of Related Literature .................................. 2 C. Purpose of Study ................................................. 26 D. Hypotheses ........................................................ 26 E. Aims of the Study ................................................ 26 MATERIALS AND METHODS ........................................................... 27 RESULTS .................................................................................... 64 DISCUSSION ................................................................................ 74 SUMMARY ....................................................................................86 GLOSSARY ......................................... : ....................................... 93 BIBLIOGRAPHY ............ ·............................. : .................................. 97 iii ACKNOWLEDGEMENTS I would like to express my sincere gratitude and appreciation to the following people who inspired, motivated, and supported me in completing this thesis. Without your insight, encouragement, and patience, this product could not have been realized. First and foremost, to my wife Patricia, you are the rock of my life and the reason that I do all that I do. Thank you for your extreme patience and fortitude in maintaining our home during the past three years. You are truly the nucleus of our family unit. Your personal discipline and support have served me well, and helped me to remain steadfast in the accomplishment of all required tasks. I will remain in your debt eternally. I love you. To my children Bianca, Ricky and Hannah, thank you all for the patience and understanding displayed, in the pursuit of this long-term goal and educational project. I was absent during many special events in your lives. I will strive to be as patient with you in your future endeavors, as you were with me. To my major advisor Dr. Mohamed Sharawy, thank you for providing the necessary guidance and mentorship during this project. You have been a true inspiration for me, as I aspire to become an anatomist, surgeon, and scientist. I will forever consider you a mentor and role model. Thank you for showing me the higher purpose and true reward to be found in a research career. To Dr. David Pashley and Dr. James Barke, thank you for your support and guidance as members of my graduate research committee. Your input has been essential in helping me realize the potential for multiple answers to scientific questions. To Dr. Frederick Rueggeberg and Dr. Gretchen Caughman, thank you for guiding me through the creation of the thesis proposal and helping me develop a confident scientific writing style. To Mrs. Linda Sterling, thank you for all your administrative support and help in coordinating interaction with the major advisor and graduate research committee. iv To Dr. Gary Swiec, thank you for your support and guidance during the last three years. You were the inspiration for me to pursue the surgical specialty of periodontics. Through you, I was able to see the broad career opportunities that were possible with this specialty. Thank you for challenging me, and helping me to develop as a surgeon and scientist. To Dr. James McPherson, Ill, thank you for helping me develop an original research investigation, and helping me understand the dynamics of the experiment. Your support, guidance, and creativity were absolutely pivotal in the accomplishment of this endeavor. To Mr. Royce Runner, the center of my laboratory success rests with you and your dedication. Thank you for your technical mentorship and most especially, your patience. You were always willing to stop your own projects to lend a much needed helping hand. Because of your selfless efforts, I was able to produce a quality scientific product. I am eternally grateful. To Dr. Salwa Kirbah, Ms. Vera Larke, and Ms. Cheryl Mims, thank you all for the support, spiritual uplifting, and technical expertise and guidance you offered me in developing my laboratory protocols and techniques. Thank you for allowing me to develop independent skills, while giving me the maternal nudge when needed. Because of your tutelage and patience, I am prepared for the next step in my research career. I will always remember you. To Mr. Eugene Cauley, and the Clinical Investigations Staff, Dwight D. Eisenhower Army Medical Center, because of your knowledge and special talents in maintaining the experimental animals, accounting for resources, and managing the surgical suite, I was able to complete this study. I will always be grateful and mindful of your patience and flexibility. You all are truly the epitome of professionalism and teamwork. To Dr. Frederick Bisch and Dr. Robert Herold, thank you for all your guidance and for teaching me the art of surgical periodontics. Your inspirational zeal for the profession, will serve as a model as I develop my own pedagogical style. Your enthusiasm, for the specialty, was refreshing and has allowed me to develop a broader scope of thinking in planning treatment for our patients. To Dr. Mark Peacock, thank you for the your guidance and mentors hip in clinical periodontics. Also, thank you for your motivation and support in my decision to pursue a dental research career. Your example of versatility shall serve as a constant reminder of the flexibility necessary for successful leadership. v To Dr. Steven Hokett, thank you for helping me design this research project and training me for the surgical phase of the experiment. To Dr. David L. Carr, thank you for encouraging me to pursue the research track and follow my true professional interests. Your diverse career as an educator, clinician, administrator, and military commander will serve as a template for me, as I aspire toward a distinguished record of service. To Dr. Jose Pizarro-Matos, thank you for helping me with the image analysis programs. To Dr. Thomas Buxton, thank you for your guidance in the statistical analysis and data comparison for this study. To Dr. Henry Chuang, thank you for encouraging me to pursue a career in scientific investigation. To Ms. Vickie Browning, thank you for keeping me abreast of all the requirements of the School of Graduate Studies, Medical College of Georgia and supporting me in this educational endeavor. You are an invaluable asset to our Education Department. Finally, to Ms. Cheryl Milford, Ms. Karon Byrd, Mrs. Lynn McTier, and Mrs. Renate Blume, thank you all for serving as my dental assistants during the planning, preparation, and execution of my study. Your diligence and conscientiousness made patient management and scheduling seem effortless. Your selfless service was critical to the success of my clinical training. Thank you for helping me develop and hone my surgical skills, and thank you for your friendship. I will miss you all. vi LIST OF FIGURES Figure Page 1 Norian® CRS® Fast Set™ Putty ............................................... 14 2 Armamentarium for Critical Size Defect Surgical Protocol.. ............. 32 3 Life Net DFDBA ..................................................................... 32 4 8-mm Diameter Craniotomy ..................................................... 33 5 Critical Size Defect in Rattus norveqicus ..................................... 33 6 Ectodurally Placed PTFE Membrane ..........................................34 7 CSD with DFDBA Placed ........................................................34 8 Stereotaxic Device ................................................................. 35 9 Rat Placed in Stereotaxic Device with Scavenger ......................... 35 10 Closed Wound with Everted Margins ......................................... 36 11 Quadrangular Section of Cranial Bone ....................................... 36 12 Decalcification Solution with Bone Tissue in Cartridges ..................42 13 Technicon Automatic Tissue Processor ...................................... 43 14 Reichert-Jung Tissue Embedding Center ....................................44 15 Microtome for Paraffin Sections ................................................44 16 WELL Diamond Wire Saw ......................................................