Hydroxyapatite Degradation and Biocompatibility
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HYDROXYAPATITE DEGRADATION AND BIOCOMPATIBILITY DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree of Philosophy in the Graduate School of The Ohio State University By Haibo Wang, B.S. * * * * * The Ohio State University 2004 Dissertation Committee: Approved by Professor John Lannutti, Adviser Professor Henk Verweij _____________________________ Adviser Professor Derek Hansford Materials Science and Engineering ABSTRACT Hydroxyapatite (HA) is widely used as a bioactive ceramics since it forms a chemical bonding to bone. The disadvantage of this material is its poor mechanical properties. HA can be degraded in body, which is the reason for its bioactivity, but too fast degradation rate could cause negative effects, such as macrophage present, particle generation, and even implant clinical failure. HA degradation rate will be greatly changed under many conditions: purity, HA form (i.e. bulk form, porous form, coating, or HA/polymer composites), microstructure, implant site, body conditions, etc. Although much work has been done in HA properties and application areas, the HA degradation behavior and mechanism under these different conditions are still not clear. In this research, three aspects of HA degradation have been studied: 1) Two very common impurities—Tri-Calcium Phosphate (TCP) and Calcium Oxide and their influences on HA degradation in vitro and in vivo, 2) influence of HA/polymer composite form on HA degradation, 3) HA material particle generation and related mechanism. From the in vitro and in vivo tests on bulk HA disks with various Ca/P ratios, HA degradation can clearly be found. The degradation level is different in different Ca/P ratio samples as well as in different test environments. In same test environment, non- stoichiometric HA samples have higher degradation rate than stoichiometric HA. HA/PMMA composite design successfully intensifies HA degradation both in vitro and ii in vivo. Grain boundary damage can be found on in vivo test samples, which has not been clearly seen on bulk HA degraded surface. HA particle generation is found in in vitro and in vivo HA/PMMA composite surface and in vivo bulk HA surface. Sintering temperature and time does affect HA grain size, and this affect HA degradation rate. Intergranular fracture is found in a several micron zone close to the Ca/P ratio 1.62 and 1.67 sample degraded surfaces. At Ca/P ratio greater than 1.667, after HA degradation in water, solution pH increases because of CaO presence. iii Dedicated to my wife iv ACKNOWLEDGMENTS I want to thank my adviser, Dr. John J. Lannutti, for his intellectual guidance, encouragement and incredible patience which made this thesis possible. I thank Dr. David Anderson from Veterinary Clinical Sciences, who give us the opportunity to do in vivo tests. I also thank Dr. Amr Moursi and Phillip Winnard for their help on osteoblastic cell culture and the in vitro test. Special thanks go to Dr. Henk Verweij and Dr. Derek Hansford for serving on my defense committee and providing valuable suggestions to the completion of this work. I am especially grateful to Dr. Jong-Kook Lee-the visiting professor from Chosun Univerisity, Kwangju, Korea, who joined this research and during the one year on his staying with us, he gave us important contribution on the literature review. I wish to thank all the members of our research group for providing a joyful working site, especially Kathy Lu, Nan Guo, Shiling Ruan, J. Dawson White, Wenxia Li, Taryn Sproule, Jin Nam, Heather Power, Zhijun Zhao, Alex Tsai, Andrew Kohm, and Kathy Elias. I thank my grandparents, my mother for their constant love and support to me. Finally, my loving appreciation goes to my wife Ying, who encouraged me and unconditionally supported me. v VITA 1972 ……………………………………… Wanshan, GuiZhou Province, China. 1995 ……………………………………… B.S. Mechanical Engineering, Tsinghua Univ. Beijing, China. 1998 .............................................................. Research Assistant, National Ceramic Lab, Beijing, China. 1999 - present ............................................... Graduate Teaching and Research Associate, The Ohio State University PUBLICATIONS Research Publication 1. Wang, Haibo, Jong-Kook Lee, Amr Moursi, John J. Lannutti. “Microstructural disassembly of calcium phosphates.” Journal of Biomedical Materials Research, Vol. 68A, p61-70, 2004. 2. Wang, Haibo, Jong-Kook Lee, Amr Moursi, David Anderson, Phillip Winnard, Heather Powell, John Lannutti. “Ca/P ratio effects on the degradation of hydroxyapatite in vitro.” Journal of Biomedical Materials Research, Vol. 67A, p599 - 608, 2003. 3. Wang, Haibo, Jiemo Tian. “Improvement of ZrO2 Ceramic Performance by Coating Small Amount CeO2 On 3mol%Y2O3-ZrO2 Powder.” 1st International High-Performance Ceramic Conference in Beijing, China. 1998. FIELDS OF STUDY Major Field: Materials Science and Engineering vi TABLE OF CONTENTS Page Abstract ………………………………………………………………….ii Dedication ..................................................................................................... iv Acknowledgments ..................................................................................................... v Vita ..................................................................................................... vi List of Tables ..................................................................................................... ix List of Figures ..................................................................................................... x Chapters: 1. Introduction ..................................................................................................... 1 1.1 The advantages and disadvantages of HA material................................ 2 1.2 Preparation of stoichiometric HA powders ............................................ 5 1.2.1 Syntheses based on theoretical compositions………………….5 1.2.2 Equilibrium syntheses in solution............................................... 7 1.2.3 Miscellaneous methods............................................................... 8 1.3 HA dissolution behavior and mechanism ............................................... 8 1.3.1 non-biological HA dissolving behavior in water and acid......... 9 1.3.2 Biology-related HA dissolving ................................................... 16 1.4 The unresolved issues and importance of study on HA degradation..... 21 1.5 Practical issues which influence HA degradation .................................. 23 1.5.1 HA impurity ................................................................................ 23 1.5.2 HA form of HA composites........................................................ 30 1.5.3 HA material particle generation.................................................. 36 2. Ca/P ratio effects on the degradation of hydroxyapatite in vitro ....................... 48 2.1 Introduction ............................................................................................. 48 2.2 Experiment design................................................................................... 50 2.3 Results .....................................................................................................52 2.4 Discussion ...............................................................................................54 2.4.1 Weight gain/surface precipitation............................................... 57 2.4.2 Potential in vivo significance ...................................................... 58 2.5 Conclusions ............................................................................................. 59 3 In vivo degradation results and microstructural disassembly............................. 86 vii 3.1 Introduction ............................................................................................. 86 3.2 Experiment............................................................................................... 88 3.2.1 Preparation of pure and biphasic HA.......................................... 88 3.2.2 In vitro sample preparation ......................................................... 90 3.2.3 Exposure to primary osteoblast culture ...................................... 91 3.2.4 Subcutaneous implantation ......................................................... 93 3.2.5 Bone implantation ....................................................................... 93 3.2.6 Specimen Harvest........................................................................ 94 3.3 Results .....................................................................................................95 3.3.1 In vivo study under skin and in bone on bulk HA disks............. 95 3.3.2 In vivo bone apposition and subcutaneous tests on HA/PMMA composite..................................................................................... 96 3.3.3 Grain pullout in vitro................................................................... 97 3.3.4 Grain pullout in vivo.................................................................... 98 3.3.5 Pure HA degradation in vivo....................................................... 100 3.4 Discussion................................................................................................ 100 3.5 Conclusions ............................................................................................. 104 4 Further study on hydroxyapatite degradation ..................................................... 138 4.1 Introduction ............................................................................................