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The use of low intensity pulsed ultrasound and mesenchymal stem cells in enhancing spinal fusion: --an in vitro and in vivo study.

Hui, Fan Fong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 153-181). / Abstract also in Chinese. / Acknowledgements --- p.ii / Abstract --- p.iii / Abbreviations --- p.vii / Table of Contents --- p.ix / List of Tables --- p.xv / List of Tables --- p.xv / List of Figures --- p.xvi / Major Conference Presentations --- p.xix / Publications in Preparation --- p.xxii / Chapter Chapter 1 --- Study Background --- p.1 / Chapter 1. --- Introduction --- p.2 / Chapter 1.1. --- Spinal Deformities --- p.2 / Chapter 1.1.1. --- Treatment --- p.2 / Chapter 1.2. --- Spinal fusion --- p.4 / Chapter 1.2.1. --- Gold Standard of Spinal Fusion --- p.4 / Chapter 1.2.2. --- Decortication in Spinal Fusion --- p.4 / Chapter 1.2.3. --- Autograft in Spinal Fusion --- p.4 / Chapter 1.2.4. --- Local Factors Influencing Spinal Fusion --- p.5 / Chapter 1.2.5. --- Ultimate Goals of Spinal Fusion --- p.7 / Chapter 1.2.6. --- Limitations of Spinal fusion --- p.7 / Chapter 1.3. --- Alternatives of Different Components for Enhancing Spinal Fusion / Chapter 1.3.1. --- Bone Graft Substitute --- p.9 / Chapter 1.3.2. --- Bioactive Factors --- p.15 / Chapter 1.4. --- Limitations of the Alternative Methods in Spinal Fusion Enhancement --- p.19 / Chapter 1.4.1. --- BMPs --- p.19 / Chapter 1.4.2. --- Gene Therapy --- p.20 / Chapter 1.4.3. --- Biophysical Stimulation --- p.20 / Chapter 1.5. --- Recent Methods in Enhancing Spinal Fusion --- p.21 / Chapter 1.5.1. --- Low Intensity Pulsed Ultrasound --- p.21 / Chapter 1.5.2. --- Mesenchymal Stem Cells in Spinal Fusion --- p.24 / Chapter 1.6. --- Conclusion --- p.26 / Chapter Chapter 2 --- "Hypothesis, Objectives and Plan of Study" --- p.29 / Chapter 2. --- "Hypothesis, Objectives and Plan of Study" --- p.30 / Chapter 2.1 --- Study Hypothesis --- p.31 / Chapter 2.2 --- Study Objectives --- p.31 / Chapter 2.3 --- Plan of Study --- p.32 / Chapter 2.3.1 --- For First Objective --- p.32 / Chapter 2.3.2 --- For Second Objective --- p.32 / Chapter 2.3.3 --- For Third Objective --- p.33 / Chapter Chapter 3 --- In vitro Study of Effect of Low Intensity Pulsed Ultrasound on Mesenchymal Stem Cells --- p.34 / Chapter 3.1. --- Introduction --- p.35 / Chapter 3.2. --- Materials and Methods --- p.36 / Chapter 3.2.1. --- Experimental Animal --- p.36 / Chapter 3.2.2. --- Materials and Reagents --- p.36 / Chapter 3.2.2.1. --- Dulbecco,s Modified Eagle Medium (DMEM) --- p.36 / Chapter 3.2.2.2. --- Phosphate Buffered Saline (PBS) --- p.37 / Chapter 3.2.2.3. --- Osteogenic Medium (OS) --- p.37 / Chapter 3.2.2.4. --- Alkaline Phosphatase (ALP) Buffer --- p.37 / Chapter 3.2.2.5. --- ALP Substrate Buffer --- p.38 / Chapter 3.2.2.6. --- MTT Stock Solution --- p.38 / Chapter 3.2.2.7. --- MTT Working Solution --- p.38 / Chapter 3.2.2.8. --- Lysis buffer --- p.38 / Chapter 3.2.2.9. --- Alkaline Phosphatase (ALP) Working Reagents --- p.39 / Chapter 3.2.3. --- Isolation of Bone Marrow Derived Mesenchymal Stem Cells (BM derived MSCs) --- p.39 / Chapter 3.2.4. --- In vitro Low Intensity Pulsed Ultrasound Treatment --- p.40 / Chapter 3.2.4.1. --- In vitro LIPUS Devices --- p.40 / Chapter 3.2.4.2. --- Treatment Procedure and Experimantal Groupings --- p.40 / Chapter 3.2.5. --- Effect of LIPUS on Cell Viability and Osteogenesis in bone marrow derived-MSCs --- p.41 / Chapter 3.2.5.1. --- Cell Viability Assay --- p.41 / Chapter 3.2.5.2. --- Alkaline Phosphatase (ALP) Enzyme Activity --- p.42 / Chapter 3.2.5.3. --- Cell Morphology and Alkaline Phosphatase Cytochemistry --- p.42 / Chapter 3.2.6. --- Statistical Analysis --- p.43 / Chapter 3.3. --- Results --- p.43 / Chapter 3.3.1. --- Morphology --- p.43 / Chapter 3.3.2. --- Total Number of Viable Cells --- p.44 / Chapter 3.3.3. --- ALP Activity Absorbance --- p.44 / Chapter 3.3.4. --- ALP staining --- p.45 / Chapter 3.3.5. --- Qualitative Analysis --- p.45 / Chapter 3.3.6. --- Quantitative Analysis --- p.46 / Chapter 3.4. --- Discussion --- p.46 / Chapter 3.4.1. --- LIPUS have No Enhancing Effect on Proliferation of MSCs in Basal Medium Nor Osteogenic Medium --- p.47 / Chapter 3.4.2. --- LIPUS Stimulate Proliferation of MSCs in Early Period --- p.49 / Chapter 3.4.3. --- LIPUS Further Enhanced Osteogenesis of MSCs in Osteogenic Medium --- p.49 / Chapter 3.4.4. --- 10 mins LIPUS treatment for 7 days can positively enhance osteogenic differentiation --- p.50 / Chapter 3.4.5. --- Optimum Conditions of LIPUS was Cell Type Dependent --- p.51 / Chapter 3.4.6. --- LIPUS Promoted Osteogenesis in MSCs through Accelerated Mineralization --- p.52 / Chapter Chapter 4 --- Enhancement of Posterior Spinal Fusion The Effect of Tissue-Engineered MSC and Calcium Phosphate Ceramic composite treated with LIPUS in Vivo --- p.68 / Chapter 4.1. --- Introduction --- p.69 / Chapter 4.1.1. --- TCP Biomaterials --- p.70 / Chapter 4.2. --- Materials and Methods --- p.71 / Chapter 4.2.1. --- Materials and Reagents --- p.71 / Chapter 4.2.2. --- Preparation of MSC Derived Osteogenic Cells-tricalcium Phosphate Ceramics Composite --- p.73 / Chapter 4.2.3. --- Posterior Spinal Fusion Surgery --- p.74 / Chapter 4.2.4. --- In vivo LIPUS treatment --- p.75 / Chapter 4.2.5. --- Assessment of Fusion Mass --- p.76 / Chapter 4.2.6. --- Histology --- p.77 / Chapter 4.2.7. --- Statistical Analysis --- p.79 / Chapter 4.3. --- Results --- p.79 / Chapter 4.3.1. --- Fusion by Manual Palpation --- p.79 / Chapter 4.3.2. --- pQCT Analysis --- p.80 / Chapter 4.3.3. --- Histological Analysis --- p.81 / Chapter 4.4. --- Discussion --- p.85 / Chapter 4.4.1. --- Summary of the Findings from Different Assessment Methods --- p.85 / Chapter 4.4.2. --- Addition of MSCs to TCP ceramic in Spinal Fusion --- p.87 / Chapter 4.4.3. --- The Needs of Differentiated MSC in Spinal Fusion --- p.89 / Chapter 4.4.4. --- bFGF Masked the Effect of OS in MSC --- p.91 / Chapter 4.4.5. --- LIPUS Enhanced Bone Formation --- p.95 / Chapter 4.4.6. --- LIPUS Enhanced Bone Formation through Mineralization --- p.96 / Chapter 4.4.7. --- LIPUS Enhanced Spinal Fusion through Bone Remodeling-induced Fusion Mass --- p.97 / Chapter 4.4.8. --- LIPUS Enhanced Bone Formation through Endochondral Ossification --- p.99 / Chapter Chapter 5 --- In Vivo Monitoring of Spinal Fusion in Animal Model with High-resolution Peripheral Quantitative Computed Tomography-A New Pilot Study --- p.122 / Chapter 5.1. --- Introduction --- p.123 / Chapter 5.2. --- Materials and Methods --- p.124 / Chapter 5.2.1. --- Animal Groupings --- p.124 / Chapter 5.2.2. --- Preparation of MSC Derived Osteogenic Cells-tricalcium Phosphate Ceramics Composite --- p.124 / Chapter 5.2.3. --- Posterior Spinal Fusion Operation Procedures --- p.125 / Chapter 5.2.4. --- LIPUS treatment --- p.125 / Chapter 5.2.5. --- High-resolution Peripheral Quantitative Computed Tomography …- --- p.125 / Chapter 5.2.6. --- Analysis with HR-pQCT --- p.126 / Chapter 5.3. --- Result --- p.128 / Chapter 5.3.1. --- Qualitative Observations from HR-pQCT Images --- p.128 / Chapter 5.3.2. --- Quantitative Analysis --- p.129 / Chapter 5.4. --- Discussion --- p.130 / Chapter Chapter 6 --- "Overall Summary, Discussion and Conclusion" --- p.140 / Chapter 6.1. --- Overall Summary and Discussion --- p.141 / Chapter 6.2. --- Limitations and Further Studies --- p.145 / Chapter 6.3. --- Conclusions --- p.147 / Chapter 6.4. --- Summary Flowchart of the whole thesis --- p.148 / References --- p.153

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326923
Date January 2009
ContributorsHui, Fan Fong., Chinese University of Hong Kong Graduate School. Division of Orthopaedics and Traumatology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xxii, 184 leaves : ill. (some col.) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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