Establishment of animal model for studying treatment efficacy of low-intensity pulsed ultrasound stimulations for accelerating bone-tendon repair. Standard partial patellectomy was conducted in the 18-week old rabbits that were then divided into the LIPUS treatment and control groups. The animals were followed for 2, 4, 8, and 16 weeks for various tissue analyses. LIPUS was applied to the experimental animals from postoperative day 3 to 16 weeks. We demonstrated that the healing process of PPT junction was initiated through endochondral ossification. The results showed that the size and length of newly formed bone, and its bone mineral content (BMC), but not its bone mineral density (BMD) were correlated with the failure load, ultimate strength and energy at failure. Using radiographic, biomechanical, histomorphologic and biomechanical methods, it was found that LIPUS had significant accelerating effect on PPT junction repair. We validated our study hypothesis in that LIPUS enhances bone-tendon junction healing by stimulating angiogenesis, chondrogenesis and osteogenesis. / Establishment of in vitro model for mechanism study on effects of low-intensity pulsed ultrasound stimulations. An in vitro model of osteoblast-like cell line (SaOS-2 cells) was studied using cDNA microarray to explore the molecular mechanism mediated by LIPUS. This microarray analysis revealed a total of 165 genes that were regulated at 4 and 24 hours by LIPUS treatment in osteoblastic-like cells. These genes belonged to more than ten protein families based on their function and were involved in some signal transduction pathways. This study has validated the hypothesis that LIPUS can regulate a number of critical genes transient expressions in osteoblast cell line Saos-2. / Keywords. partial patellectomy model; bone-tendon junction repair; low intensity pulsed ultrasound stimulations (LIPUS); gene expression; complementary DNA microarray; rabbit. / This study explored the intact morphology, regular healing and the augmented healing under the effects of low intensity pulsed ultrasound stimulations (LIPUS) on the patella-patella tendon (PPT) junction in a rabbit partial patellectomy model. To probe its possible mechanism, the key genes involved in regulating osteogenesis mediated by LIPUS were identified using the state-of-the-art methods---complementary DNA microarray. / Lu Hongbin. / "June 2006." / Advisers: Ling Qin; Kwok Sui Leung. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1548. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 259-288). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

by Warren Tak-keung Lee. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 161-186). / Chapter CHAPTER 1: --- INTRODUCTION --- p.1 / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Objectives --- p.3 / Chapter CHAPTER 2: --- LITERATURE REVIEW --- p.4 / Chapter 2.1 --- Calcium intakes of Hong Kong Chinese: past and present --- p.4 / Chapter 2.1.1 --- Adults --- p.4 / Chapter 2.1.2 --- Children --- p.5 / Chapter 2.2 --- Calcium Metabolism --- p.6 / Chapter 2.2.1 --- Calcium and bone mass --- p.9 / Chapter 2.2.2 --- Ethnicity and bone mass --- p.9 / Chapter 2.2.3 --- Physical activity and bone mass --- p.10 / Chapter 2.2.4 --- Hormonal control of calcium metabolism --- p.11 / Chapter 2.2.5 --- Intestinal calcium absorption --- p.14 / Chapter (1) --- Calcium transport across the intestine / Chapter (2) --- Active Calcium Transport / Chapter (3) --- Passive calcium transport / Chapter (4) --- Age and calcium absorption / Chapter 2.2.6 --- Dietary components in relation to calcium bioavailability and absorption --- p.17 / Chapter (1) --- Habitual calcium intake / Chapter (2) --- Vitamin D nutritional status / Chapter (3) --- Protein / Chapter (4) --- Phosphorous and Ca:P ratio / Chapter (5) --- Sodium / Chapter (6) --- Lactose / Chapter (7) --- Glucose and Glucose Polymers / Chapter (8) --- Phytate / Chapter (9) --- Oxalate / Chapter (10) --- Plant estrogen (phyto-estrogen) / Chapter 2.2.7 --- Bioavailability from calcium salts --- p.23 / Chapter 2.3 --- Calcium intakes and requirements --- p.25 / Chapter 2.3.1 --- Calcium requirements in adulthood --- p.28 / Chapter 2.3.2 --- Calcium requirements in childhood --- p.29 / Chapter 2.3.3 --- Manifestation of calcium deficiency in children --- p.30 / Chapter 2.4 --- Assessment of Dietary Intakes --- p.32 / Chapter 2.4.1 --- Food weighing method --- p.32 / Chapter 2.4.2 --- Food Recording method --- p.34 / Chapter 2.4.3 --- 24-hour dietary recall --- p.35 / Chapter 2.4.4 --- Food frequency method --- p.36 / Chapter 2.4.5 --- Dietary history method --- p.38 / Chapter 2.4.6 --- Chemical analysis of duplicate meals --- p.39 / Chapter 2.4.7 --- Photographic method --- p.40 / Chapter 2.4.8 --- Selecting suitable methods for the present study --- p.40 / Chapter 2.5 --- Food composition database --- p.41 / Chapter 2.6 --- Evaluation of bone mass in vivo --- p.43 / Chapter 2.6.1 --- Single photon absorptiometry --- p.44 / Chapter 2.6.2 --- Dual photon absorptiometry --- p.46 / Chapter 2.6.3 --- Dual energy X-ray absorptiometry --- p.47 / Chapter 2.6.4 --- Quantitative computerized tomography --- p.47 / Chapter 2.6.5 --- The techniques selected to quantify bone mass in the present study --- p.48 / Chapter 2.7 --- Measurement of intestinal calcium absorption --- p.49 / Chapter 2.7.1 --- Metabolic balance study --- p.49 / Chapter 2.7.2 --- Isotopic techniques (radioisotope or stable isotope) --- p.50 / Chapter (1) --- Radio isotope vs stable isotope / Chapter (2) --- The single-label isotope technique / Chapter (3) --- The double-label isotope technique / Chapter (4) --- "Preparation of stable isotopes for human study, and determination of stable isotopes in body fluids" / Chapter (I) --- Dosage considerations / Chapter (II) --- Intrinsic or Extrinsic labelling / Chapter (III) --- Oral and intravenous administration of isotopes / Chapter 2.7.3 --- The technique selected to determine calcium absorption in the present study --- p.60 / Chapter 2.8 --- Mass spectrometry --- p.60 / Chapter 2.8.1 --- Thermal ionization mass spectrometry --- p.60 / Chapter 2.8.2 --- Fast atom bombardment mass spectrometry --- p.61 / Chapter 2.8.3 --- Inductively coupled plasma mass spectrometry --- p.61 / Chapter 2.8.4 --- Electron impact mass spectrometry and Gas chromatography mass spectrometry --- p.62 / Chapter 2.8.5 --- Neutron activation analysis --- p.62 / Chapter 2.8.6 --- The type of mass spectrometry used to determine stable isotopic ratios in the present study --- p.63 / Chapter 2.9 --- Assessment of physical activity in children --- p.63 / Chapter 2.9.1 --- Activity questionnaire or record --- p.64 / Chapter 2.9.2 --- Direct measurement of physical activity --- p.65 / Chapter (1) --- Accelerometer / Chapter (2) --- Pedometer / Chapter (3) --- Actometer / Chapter (4) --- Video-recording / Chapter (5) --- Heart-rate recording / Chapter 2.9.3 --- Selection of a suitable physical activity assessment method --- p.67 / Chapter CHAPTER 3 --- RANDOMIZED DOUBLE-BLIND CONTROLLED CALCIUM SUPPLEMENTATON TRIALS IN RELATION TO BONE AND HEIGHT ACQUISITION IN 7-YEAR OLD CHINESE CHILDREN FROM JIANGMEN (CHINA) AND HONG KONG --- p.71 / Chapter 3.1 --- Chapter summary --- p.71 / Chapter 3.2 --- Chapter Introduction --- p.72 / PART I --- p.73 / Chapter 3.3 --- Double-blind Controlled Randomized Calcium Supplementation and Bone and Height Acquisition in Chinese Children Accustomed to Mean Calcium Intake at About 300mg/d --- p.73 / Chapter 3.3.1 --- Introduction --- p.73 / Chapter 3.3.2 --- Objectives --- p.74 / Chapter 3.3.3 --- Subject and Methods --- p.74 / Chapter 3.3.4 --- Results --- p.81 / Chapter 3.3.5 --- Discussions --- p.83 / Chapter 3.3.6 --- Conclusion --- p.84 / PART II --- p.84 / Chapter 3.4 --- Randomized double-blind controlled calcium supplementation in relation to bone mineral accretion and height increment of Hong Kong Chinese children --- p.84 / Chapter 3.4.1 --- Introduction --- p.84 / Chapter 3.4.2 --- Objectives --- p.85 / Chapter 3.4.3 --- Subjects and Methods --- p.85 / Chapter 3.3.4 --- Results --- p.91 / Chapter 3.4.5 --- Discussions --- p.95 / Chapter 3.4.6 --- Conclusion --- p.97 / Chapter 3.5 --- Comparisons of the two calcium supplementation trials from Jiangmen and Hong Kong --- p.97 / Chapter 3.6 --- Chapter Discussions --- p.99 / Chapter 3.7 --- Chapter Conclusion --- p.103 / Chapter CHAPTER 4 --- TRUE FRACTIONAL CALCIUM ABSORPTION OF CHINESE CHILDREN AND THE EFFECTS OF DOUBLE-BLIND CONTROLLED CALCIUM SUPPLEMENTATION ON CALCIUM ABSORPTION IN CHILDREN MEASURED WITH STABLE ISOTOPES (42Ca and 44Ca) --- p.117 / Chapter 4.1 --- Chapter summary --- p.117 / Chapter 4.2 --- Chapter introduction --- p.118 / PART I --- p.119 / Chapter 4.3 --- True fractional calcium absorption in Chinese children measured with stable isotopes (42Ca and 44Ca) --- p.119 / Chapter 4.3.1 --- Introduction --- p.119 / Chapter 4.3.2 --- Objectives --- p.120 / Chapter 4.3.3 --- "Subjects, Materials and Methods" --- p.120 / Chapter 4.3.4 --- Results --- p.127 / Chapter 4.3.5 --- Discussions & Conclusion --- p.131 / Part II --- p.131 / Chapter 4.4 --- Effects of double-blind controlled calcium supplementation on calcium absorption in Chinese children measured with stable isotopes (42Ca and 44Ca) --- p.131 / Chapter 4.4.1 --- Introduction --- p.131 / Chapter 4.4.2 --- Objective --- p.132 / Chapter 4.4.3 --- "Subjects, Materials and Methods" --- p.132 / Chapter 4.4.4 --- Results --- p.135 / Chapter 4.4.5 --- Discussions --- p.137 / Chapter 4.4.6 --- Conclusion --- p.139 / Chapter 4.5 --- Chapter Conclusion and Discussions --- p.140 / Chapter CHAPTER 5 --- GENERAL DISCUSSIONS AND RECOMMENDATIONS FOR FURTHER STUDY --- p.156 / Chapter 5.1 --- Ethnic differences in bone acquisition and calcium absorption --- p.156 / Chapter 5.2 --- Calcium requirements for Chinese children --- p.157 / Chapter 5.3 --- Indications for further studies --- p.158 / REFERENCES --- p.161 / APPENDIXES / PUBLICATIONS

Lai Yau Ming. / "August 2006." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 260-290). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

青少年特發性脊柱側凸(Adolescent idiopathic scoliosis , AIS)是一種複雜的脊柱三維畸形，常見於10-16 歲處於生長發育高峰期的青少年女性。儘管AIS 發生率較高並且臨床影響較大，但是到目前為止其病因未明。在眾多關於AIS 病因學的假設和理論研究中，普遍認為低骨密度是AIS 的一個重要影響因素。然而近年來對於AIS 患者低骨密度研究不足，其潛在的機制尚不明確。我們之前初步的組織學研究發現，AIS 患者的松質骨中成骨細胞功能下降，此研究為AIS中存在骨礦化異常提供了初步依據。 / 骨橋蛋白是骨組織中一種重要的非膠原細胞外基質蛋白，其在骨礦化過程中起著重要作用。近期的研究報導AIS 患者血漿中骨橋蛋白水準高於年齡匹配的正常對照。因此本研究假設AIS 患者血漿及骨組織中骨橋蛋白高於正常對照，并可能影響了骨基質的礦化，從而導致低骨密度。 / 本系列研究的第一部分旨在通過外周定量電腦斷層掃描（pQCT）明確AIS患者中皮質骨密度及松質骨密度是否均低於正常對照。pQCT 可以準確地三維評估皮質骨密度，松質骨密度及其他骨品質的相關參數。採用雙能X 線骨密度儀（DXA）測量受試者的非優勢側近端股骨面積骨密度（包括股骨頸，Ward’s 三角及大轉子）。而採用pQCT 測量受試者非優勢側橈骨遠端容積骨密度，包括皮質骨密度及松質骨密度。結果顯示AIS 患者面積骨密度，皮質骨密度及松質骨密度在不同年齡段和月經時間分組中均低於正常對照。並且AIS 與正常對照皮質骨密度的差異隨著年齡增長越來越大，而松質骨密度差異則隨著年齡增長越來越小。 / 第二部分通過顯微CT 及組織形態測定研究AIS 及正常骨組織的骨礦化及骨微結構。採用顯微CT 檢測骨組織的三維結構參數，包括材料骨密度及骨微結構。未脫鈣骨組織的切片通過Goldner’s 染色進行組織形態學測量。結果顯示AIS患者的骨體積分數，骨小梁數目，骨小梁厚度及結構模型指數與正常對照之間均無顯著差異，而材料骨密度顯著低於正常對照。組織形態學分析結果顯示AIS中低礦化骨顯著多於正常對照。 / 第三部分旨在研究AIS 及正常對照血漿中骨橋蛋白水準及其與骨密度的關係。採用酶聯吸附免疫法測量AIS 患者及年齡匹配的正常對照血漿中的骨橋蛋白水準。血漿骨橋蛋白水準與骨密度的關係採用多元回歸分析。研究結果顯示AIS 患者及正常對照血漿骨橋蛋白水平均與年齡及月經時間呈負相關。AIS 患者的血漿骨橋蛋白水準顯著高於正常對照，並且與松質骨密度呈顯著負相關。 / 本研究第四部分旨在探討骨組織中的骨橋蛋白表達與骨形態學及骨礦化指標在AIS 及正常對照中的關係。骨組織中骨橋蛋白的表達採用半定量免疫組織化學法評估。研究結果顯示在AIS 中血漿骨橋蛋白水準與骨組織中骨橋蛋白的表達呈正相關。且AIS 骨組織中骨橋蛋白的表達也顯著高於正常對照。進一步的研究發現骨組織中骨橋蛋白的表達與材料骨密度呈負相關，而與低礦化骨量呈正相關。 / 本研究明確了AIS 中骨礦化水準低於正常對照，進一步證明AIS 患者中的皮質骨及松質骨密度下降可能與骨礦化的調控異常有關。本研究發現的骨橋蛋白與低骨密度及低骨礦化水準的關係，可以推測AIS 患者中異常升高的骨橋蛋白水準可能在骨礦獲取的調解中起重要作用。本系列研究提供證據支援AIS 患者中骨橋蛋白的異常表達可能影響了骨基質的礦化，從而導致低骨密度。本研究為AIS 中低骨密度可能的機制提供了全新的見解，並可能進一步解釋AIS 的發病機理及其發生，發展。 / Adolescent idiopathic scoliosis (AIS) is a complex three-dimensional deformity of the spine occurring most commonly in girls between ages 10-16 during the pubertal growth spurt. Despite its high prevalence and clinical impact, etiology of AIS remains largely unknown. Among the number of proposed hypothesis and observations on the etiopathogenesis of AIS, low bone mineral density (BMD) is one of the most reported factor (Cheng et al. 1999; Hung et al. 2005; Cheung et al. 2006; Hui et al. 2011). However, the underlying mechanism of low BMD in AIS has not been sufficiently studied scientifically and its link to the etiopathogenesis is still not clear. From a previous pilot study, our group has reported the histological features of reduced osteoblastic activity in bone biopsy specimens obtained from AIS subjects intraoperatively, thus providing the early evidence of abnormal bone mineral acquisition and mineralization (Cheng et al. 2001). / Osteopontin (OPN) has been recognized as one the major non-collagen extracellular matrix proteins in bone and plays an important role in bone mineralization. Recent report suggested that AIS patients have higher OPN level than normal controls (Moreau et al. 2009). It was hypothesized that the low BMD in AIS is associated with abnormal bone matrix mineralization which may be related to abnormal expression of OPN in the plasma and at tissue level. / In this series of studies, the first part aimed to investigate the differential cortical and trabecular bone mineral density of AIS Vs normal controls. The non-dominant proximal femur areal BMD (aBMD) (femoral neck, Ward’s triangle and greater trochanter) of the subjects were measured with dual-energy x-ray absorptiometry (DXA). The volumetric bone mineral density (vBMD) in non-dominant distal radius was measured with peripheral quantitative computed tomography (pQCT) that allows accurate three dimensional assessment of the cortical and trabecular bone mineral density and other parameters of bone quality. AIS was found to have lower aBMDs, trabecular BMD (TBMD) and cortical BMD (CBMD) in different age groups and year since menarche (YSM) groups. Furthermore, the percentage difference of CBMD between AIS and controls was increased with age while a decreasing trend was observed in the TBMD. / The second part of the study investigated the bone mineralization and bone micro-architecture with micro-computed tomography (micro-CT) and histomorphometry study of bone biopsies obtained from AIS and normal controls. Three-dimensional structural parameters including material bone mineral density (mBMD) and bone architecture were evaluated by micro-CT. Bone histomorphometry was assessed by undecalcified sectioning with Goldner’s trichrome staining. mBMD of trabecular bone in AIS was found to be significantly lower than the normal control while no difference could be demonstrated in BV/TV, Tb.N, Tb.Th and SMI measurement between the two groups. It was also shown that the percentage of low-mineralized bone in AIS was significantly higher than that in normal controls. / The third part aimed to study the plasma OPN level and its association with the BMD in AIS Vs normal controls. Plasma OPN level in AIS and age-matched controls was measured by ELISA. With multivariate regression analysis, the plasma OPN level was found to be negatively correlated with Age and YSM in both AIS and normal controls. In addition, the plasma OPN level in AIS was significantly higher and correlated with the low trabecular BMD. / The fourth part of the study investigated the OPN expression in bone tissues level and its association with histomorphometric bone mineralization and bone micro-architectural parameters in AIS Vs normal controls. OPN expression in bone biopsy was semi-quantified by immunohistochemistry. It was found that the bone tissue OPN level was significantly higher in AIS and also positively correlated with plasma OPN level. In addition, in this pilot study, we found the trend that OPN expression in trabecular bone was negatively associated with mBMD, and positively with the percentage of low-mineralized bone. / The present study showed that AIS had lower bone mineralization than normal controls. The low cortical and trabecular BMD found in AIS is likely to be resulting from abnormal regulation of bone mineralization. The association of OPN with abnormal BMD and bone mineralization further suggested that abnormal OPN level might play an important role in affecting the bone mineral acquisition in AIS. All of these findings strongly supported the hypothesis that the low BMD in AIS is associated with abnormal bone matrix mineralization which could be related to abnormal expression of OPN. This study provided important additional insight into the possible mechanism of lower bone mineral density that might be linked to theetiopathogenesis, development and progression of the spinal deformity in AIS. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Sun, Guangquan. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 143-160). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract and appendix A also in Chinese. / THE CHINESE UNIVERSITY OF HONG KONG --- p.I / ACKNOWLEDGEMENTS --- p.II / ABSTRACT --- p.IV / ABBREVIATION --- p.XI / TABLE OF CONTENTS --- p.XIII / LIST OF TABLES --- p.XVII / LIST OF FIGURES --- p.XIX / LIST OF PUBLICATIONS --- p.XXI / Chapter CHAPTER 1 --- STUDY BACKGROUND --- p.1 / Chapter 1.1 --- GENERAL OVERVIEW OF ADOLESCENT IDIOPATHIC SCOLIOSIS (AIS) --- p.2 / Chapter 1.1.1 --- NATURAL HISTORY --- p.4 / Chapter 1.1.2 --- CURRENT TREATMENTS --- p.6 / Chapter 1.1.2.1 --- Observation --- p.7 / Chapter 1.1.2.2 --- Bracing --- p.7 / Chapter 1.1.2.3 --- Surgical treatments --- p.9 / Chapter 1.1.3 --- CURRENT HYPOTHESIS ON THE ETIOLOGY OF AIS --- p.11 / Chapter 1.1.3.1 --- Genetic factors --- p.12 / Chapter 1.1.3.2 --- Neuromuscular impairment --- p.14 / Chapter 1.1.3.3 --- Abnormalities in skeletal development --- p.16 / Chapter 1.1.3.4 --- Low bone mineral density in AIS --- p.16 / Chapter 1.2 --- BONE MINERALIZATION --- p.18 / Chapter 1.2.1 --- Overview of bone mineralization --- p.18 / Chapter 1.2.2 --- Bone modeling --- p.18 / Chapter 1.2.3 --- Bone remodeling --- p.19 / Chapter 1.2.4 --- Factors affecting bone mineralization --- p.21 / Chapter 1.3 --- OSTEOPONTIN --- p.23 / Chapter 1.3.1 --- Structure of osteopontin --- p.23 / Chapter 1.3.2 --- Osteopontin - cellular and tissue distribution --- p.24 / Chapter 1.3.3 --- Osteopontin functions --- p.25 / Chapter 1.3.4 --- Osteopontin functions in bone --- p.25 / Chapter 1.3.5 --- Osteopontin and bone mineral density in human --- p.29 / Chapter CHAPTER 2 --- STUDY HYPOTHESIS AND PLAN --- p.31 / Chapter 2.1 --- INTRODUCTION --- p.32 / Chapter 2.2 --- HYPOTHESIS --- p.33 / Chapter 2.3 --- OBJECTIVES --- p.34 / Chapter 2.4 --- STUDY PLAN --- p.34 / Chapter CHAPTER 3 --- LOW BONE MINERAL DENSITY IN ADOLESCENT IDIOPATHIC SCOLIOSIS - AREAL VS VOLUMETRIC, CORTICAL VS TRABECULAR BONE MINERAL DENSITY --- p.36 / Chapter 3.1 --- INTRODUCTION --- p.37 / Chapter 3.2 --- SUBJECTS AND METHODS --- p.39 / Chapter 3.2.1 --- Subjects --- p.39 / Chapter 3.2.2 --- BMD Measurement --- p.40 / Chapter 3.2.3 --- Statistical Analysis --- p.41 / Chapter 3.3 --- RESULTS --- p.42 / Chapter 3.3.1 --- aBMD of AIS and normal controls by age groups --- p.42 / Chapter 3.3.2 --- TBMD and CBMD in AIS and normal controls by age groups --- p.42 / Chapter 3.3.3 --- aBMD in AIS and normal controls by year since menarche --- p.43 / Chapter 3.3.4 --- TBMD and CBMD in AIS and normal controls by year since menarche --- p.43 / Chapter 3.3.5 --- Correlation between CBMD & TBMD and chronological age or year since menarche --- p.44 / Chapter 3.3.6 --- Comparisons adjusted for chronological age or year since menarche --- p.44 / Chapter 3.4 --- DISCUSSION --- p.45 / Chapter 3.5 --- TABLES AND FIGURES --- p.50 / Chapter CHAPTER 4 --- ABNORMAL BONE MATRIX MINERALIZATION AND BONE MICROARCHITECTURE IN ADOLESCENT IDIOPATHIC SCOLIOSIS - A HISTOMORPHOMETRIC AND MICRO-CT STUDY --- p.60 / Chapter 4.1 --- INTRODUCTION --- p.61 / Chapter 4.2 --- SUBJECTS AND METHODS --- p.62 / Chapter 4.2.1 --- Subjects --- p.62 / Chapter 4.2.2 --- Micro-computed tomography --- p.63 / Chapter 4.2.3 --- Bone histomorphometry --- p.64 / Chapter 4.2.4 --- Statistical analysis --- p.68 / Chapter 4.3 --- RESULTS --- p.68 / Chapter 4.3.1 --- Results of micro-CT analysis --- p.68 / Chapter 4.3.2 --- Results of histomorphometric analysis --- p.69 / Chapter 4.3.3 --- Relationship of mBMD and percentage of low-mineralized bone --- p.69 / Chapter 4.4 --- DISCUSSION --- p.70 / Chapter 4.5 --- TABLES AND FIGURES --- p.74 / Chapter CHAPTER 5 --- PLASMA OSTEOPONTIN LEVEL AND ITS ASSOCIATION WITH BONE MINERAL DENSITY IN ADOLESCENT IDIOPATHIC SCOLIOSIS --- p.82 / Chapter 5.1 --- INTRODUCTION --- p.83 / Chapter 5.2 --- SUBJECTS AND METHODS --- p.84 / Chapter 5.2.1 --- Subjects --- p.84 / Chapter 5.2.2 --- Anthropometric assessment --- p.84 / Chapter 5.2.3 --- Plasma osteopontin measurement --- p.85 / Chapter 5.2.4 --- BMD Measurement --- p.86 / Chapter 5.2.5 --- Statistical Analysis --- p.86 / Chapter 5.3 --- RESULTS --- p.86 / Chapter 5.3.1 --- Comparison of anthropometric parameters between AIS and controls --- p.86 / Chapter 5.3.2 --- Correlation between OPN plasma level with age or YSM in AIS and controls --- p.87 / Chapter 5.3.3 --- Comparison of OPN plasma level between AIS and controls --- p.87 / Chapter 5.3.4 --- Correlation between OPN plasma level and curve severity in AIS --- p.87 / Chapter 5.3.5 --- Relationship between OPN plasma level and vBMD --- p.88 / Chapter 5.4 --- DISCUSSION --- p.88 / Chapter 5.5 --- TABLES AND FIGURES --- p.94 / Chapter CHAPTER 6 --- OSTEOPONTIN EXPRESSION IN BONE TISSUE AND ITS ASSOCIATION WITH BONE MATRIX MINERALIZATION IN ADOLESCENT IDIOPATHIC SCOLIOSIS - A PILOT STUDY --- p.102 / Chapter 6.1 --- INTRODUCTION --- p.103 / Chapter 6.2 --- SUBJECTS AND METHODS --- p.104 / Chapter 6.2.1 --- Subjects --- p.104 / Chapter 6.2.2 --- Micro-computed tomography --- p.104 / Chapter 6.2.3 --- Bone histomorphometry --- p.104 / Chapter 6.2.4 --- Semi-quantification of OPN expression in bone biopsy by immunohistochemistry --- p.105 / Chapter 6.2.5 --- Plasma osteopontin measurement --- p.107 / Chapter 6.2.6 --- Statistical Analysis --- p.108 / Chapter 6.3 --- RESULTS --- p.108 / Chapter 6.3.1 --- Comparison of anthropometric parameters between AIS and control subjects --- p.108 / Chapter 6.3.2 --- Comparison of OPN expression detected by immunohistochemistry in bone biopsy between AIS and control groups --- p.108 / Chapter 6.3.3 --- Comparison of histomorphometric and micro-CT results between AIS and control groups --- p.109 / Chapter 6.3.4 --- Relationship between plasma OPN level and OPN expression in bone biopsy --- p.109 / Chapter 6.3.5 --- Relationship between percentage of low-mineralized bone and OPN expression in bone biopsy --- p.109 / Chapter 6.3.6 --- Relationship between material bone mineral density and OPN expression in bone biopsy --- p.110 / Chapter 6.4 --- DISCUSSION --- p.110 / Chapter 6.5 --- TABLES AND FIGURES --- p.114 / Chapter CHAPTER 7 --- SUMMARY STUDY FLOWCHART, OVERALL DISCUSSION, CONCLUSIONS, LIMITATIONS AND FURTHER STUDIES --- p.119 / Chapter 7.1 --- SUMMARY OF THE STUDY FLOW CHART WITH KEY FINDINGS --- p.120 / Chapter 7.2 --- OVERALL DISCUSSION --- p.125 / Chapter 7.2.1 --- The novel findings on bone mineralization abnormality in AIS in this study --- p.125 / Chapter 7.2.2 --- OPN is a key modulator in AIS --- p.128 / Chapter 7.3 --- OVERALL CONCLUSIONS --- p.130 / Chapter 7.4 --- LIMITATION OF THIS STUDY AND FUTURE RESEARCH --- p.131 / Chapter APPENDIX A. --- CONSENT FORM OF AIS RESEARCH --- p.135 / Chapter APPENDIX B. --- CONSENT FORM OF BONE BIOPSY COLLECTION --- p.137 / Chapter APPENDIX C. --- MATERIALS AND REAGENTS INFORMATION AND PROTOCOL FOR SOLUTIONS PREPARATION --- p.138 / BIBLIOGRAPHY --- p.143

OBJETIVO: Determinar os valores séricos do ligante do receptor do ativador do fator nuclear Kappa B (RANKL) e da osteoprotegerina (OPG) em pacientes com Artrite Idiopática Juvenil de início poliarticular (AIJp) em atividade e avaliar a possível correlação destes valores com a presença radiológica de erosões ósseas. MÉTODOS: Trinta pacientes do sexo feminino com diagnóstico de AIJ de início poliarticular segundo os critérios da ILAR (International League of Associations for Rheumatology) em atividade e trinta crianças saudáveis (controles) pareadas por idade e sexo foram selecionadas consecutivamente para este estudo. Todas as articulações comprometidas foram radiografadas e avaliadas, com especial interesse, para a presença de erosões ósseas. Concentrações séricas do RANKL e OPG foram medidas por enzima-imunoensaio (Biomedica, Vienna, Austria). RESULTADOS: Pacientes com AIJp em atividade apresentaram altos valores séricos de RANKL e menores taxas de OPG/RANKL comparadas com controles [2,90 (0,1-37,4) vs. 0,25 (0,1-5,7) pg/ml, p=0,007 e 21,25 (1,8- 897,6) vs. 347,5 (9-947,8)] pg/ml, p=0,005). Diferentemente, não houve diferença em relação à concentração sérica de OPG quando se comparou os pacientes e controles [55,24 (28,34-89,76) vs. 64,42 (30,68-111,28) pg/ml, p=0,256]. Maiores concentrações de RANKL e menores taxas de OPG/RANKL também foram observadas em pacientes com AIJp em atividade com erosões ósseas comparadas com controles [3,49 (0,1-37,4) vs. 0,25 (0,1-5,7) pg/ml, p=0,0115 e 14,3 (1,8-897.6) vs. 347,5 (9-947,8) p=0,016]. Em contraste, valores séricos de RANKL e a taxa de OPG/RANKL foram semelhantes em pacientes com AIJp sem erosões ósseas comparadas com controles [1,75 (0,1-10,9) vs 0,25 (0,1-5,7) pg/ml, p=0,055 e 29,2 (3,3-756,8) vs. 347,5 (9- 947,8), p=0,281]. CONCLUSÃO: Estes dados sugerem que pacientes com AIJp em atividade com erosões ósseas apresentam altos valores séricos de RANKL e baixa taxa de OPG/RANKL indicando que estas alterações podem refletir o comprometimento ósseo nesta doença. / OBJECTIVE: To determine the serum levels of receptor activator of nuclear factor kB-ligand (RANKL) and osteoprotegerin (OPG) in active polyarticularonset Juvenile Idiopathic Arthritis patients (pJIA) and evaluate its possible correlation with bone erosions on the X-ray. METHODS: Thirty female girls with active pJIA diagnosis according ILAR criteria (International League of Associations for Rheumatology) and 30 healthy children gender and agematched controls were consecutively selected for this study. All involved articulations were evaluated by X-ray and analyzed for the presence of bone erosions. The serum levels of RANKL and OPG were measured using an enzyme-linked immunosorbent (Biomedica, Vienna, Austria). RESULTS: Results: Patients with active pJIA had higher levels of serum RANKL and lower OPG/RANKL ratio compared to controls [2.90 (0.1-37.4) vs. 0.25 (0.1- 5.7) pg/ml, p=0.007] and 21.25 (1.8-897.6) vs. 347.5 (9-947.8) pg/ml, p=0.005]. However, levels of OPG were comparable in both groups [55.24 (28.34-89.76) vs. 64.42 (30.68-111.28) pg/ml, p=0.256]. Higher levels of serum RANKL and lower OPG/RANKL ratio was also observed in active pJIA patients with bone erosions compared to controls [3.49 (0.1-37.4) vs. 0.25 (0.1-5.7) pg/ml, p=0.0115 and 14.3 (1.8-897.6) vs. 347.5 (9-947.8), p=0.016]. In contrast, RANKL levels and OPG/RANKL ratio were alike in pJIA patients without bone erosion and controls [1.75 (0.1-10.9) vs. 0.25 (0.1- 5.7) pg/ml, p=0.055 and 29.2 (3.3-756.8) vs. 347.5 (9-947.8), p=0.281]. CONCLUSION: These data suggest that active pJIA with bone erosion is associated with high serum levels of RANKL and low OPG/RANKL ratio indicating that these alterations may reflect bone damage in this disease.

OBJETIVO: Determinar os valores séricos do ligante do receptor do ativador do fator nuclear Kappa B (RANKL) e da osteoprotegerina (OPG) em pacientes com Artrite Idiopática Juvenil de início poliarticular (AIJp) em atividade e avaliar a possível correlação destes valores com a presença radiológica de erosões ósseas. MÉTODOS: Trinta pacientes do sexo feminino com diagnóstico de AIJ de início poliarticular segundo os critérios da ILAR (International League of Associations for Rheumatology) em atividade e trinta crianças saudáveis (controles) pareadas por idade e sexo foram selecionadas consecutivamente para este estudo. Todas as articulações comprometidas foram radiografadas e avaliadas, com especial interesse, para a presença de erosões ósseas. Concentrações séricas do RANKL e OPG foram medidas por enzima-imunoensaio (Biomedica, Vienna, Austria). RESULTADOS: Pacientes com AIJp em atividade apresentaram altos valores séricos de RANKL e menores taxas de OPG/RANKL comparadas com controles [2,90 (0,1-37,4) vs. 0,25 (0,1-5,7) pg/ml, p=0,007 e 21,25 (1,8- 897,6) vs. 347,5 (9-947,8)] pg/ml, p=0,005). Diferentemente, não houve diferença em relação à concentração sérica de OPG quando se comparou os pacientes e controles [55,24 (28,34-89,76) vs. 64,42 (30,68-111,28) pg/ml, p=0,256]. Maiores concentrações de RANKL e menores taxas de OPG/RANKL também foram observadas em pacientes com AIJp em atividade com erosões ósseas comparadas com controles [3,49 (0,1-37,4) vs. 0,25 (0,1-5,7) pg/ml, p=0,0115 e 14,3 (1,8-897.6) vs. 347,5 (9-947,8) p=0,016]. Em contraste, valores séricos de RANKL e a taxa de OPG/RANKL foram semelhantes em pacientes com AIJp sem erosões ósseas comparadas com controles [1,75 (0,1-10,9) vs 0,25 (0,1-5,7) pg/ml, p=0,055 e 29,2 (3,3-756,8) vs. 347,5 (9- 947,8), p=0,281]. CONCLUSÃO: Estes dados sugerem que pacientes com AIJp em atividade com erosões ósseas apresentam altos valores séricos de RANKL e baixa taxa de OPG/RANKL indicando que estas alterações podem refletir o comprometimento ósseo nesta doença. / OBJECTIVE: To determine the serum levels of receptor activator of nuclear factor kB-ligand (RANKL) and osteoprotegerin (OPG) in active polyarticularonset Juvenile Idiopathic Arthritis patients (pJIA) and evaluate its possible correlation with bone erosions on the X-ray. METHODS: Thirty female girls with active pJIA diagnosis according ILAR criteria (International League of Associations for Rheumatology) and 30 healthy children gender and agematched controls were consecutively selected for this study. All involved articulations were evaluated by X-ray and analyzed for the presence of bone erosions. The serum levels of RANKL and OPG were measured using an enzyme-linked immunosorbent (Biomedica, Vienna, Austria). RESULTS: Results: Patients with active pJIA had higher levels of serum RANKL and lower OPG/RANKL ratio compared to controls [2.90 (0.1-37.4) vs. 0.25 (0.1- 5.7) pg/ml, p=0.007] and 21.25 (1.8-897.6) vs. 347.5 (9-947.8) pg/ml, p=0.005]. However, levels of OPG were comparable in both groups [55.24 (28.34-89.76) vs. 64.42 (30.68-111.28) pg/ml, p=0.256]. Higher levels of serum RANKL and lower OPG/RANKL ratio was also observed in active pJIA patients with bone erosions compared to controls [3.49 (0.1-37.4) vs. 0.25 (0.1-5.7) pg/ml, p=0.0115 and 14.3 (1.8-897.6) vs. 347.5 (9-947.8), p=0.016]. In contrast, RANKL levels and OPG/RANKL ratio were alike in pJIA patients without bone erosion and controls [1.75 (0.1-10.9) vs. 0.25 (0.1- 5.7) pg/ml, p=0.055 and 29.2 (3.3-756.8) vs. 347.5 (9-947.8), p=0.281]. CONCLUSION: These data suggest that active pJIA with bone erosion is associated with high serum levels of RANKL and low OPG/RANKL ratio indicating that these alterations may reflect bone damage in this disease.

Diss. (sammanfattning) Umeå : Umeå universitet, 2009. / Härtill 4 uppsatser. Även tryckt utgåva.

FGFR 3 is one of a family of four high affinity receptors for FGF ligands. Activating mutations in FGFR 3 result in skeletal dysplasias that vary in severity from undetectable to neonatal lethal. Mice with congenital deficiency of FGFR3 develop severe kyphosis and skeletal overgrowth. FGFR3 is also expressed in calvarial pre-osteoblasts, osteoblast and articular chondrocytes, although it biological role in these cells remains undefined. By changing the genetic background of the Fgfr3-/- mice we were able to extend their lifespan and examine its impact on post-natal skeletal growth. To investigate the implication of FGFR 3 in post-natal cartilage and bone metabolism we used a combination of imaging, classic histology, molecular biology and biomechanical testing. The results demonstrated that the synovial joints of young adult Fgfr3-/- mice revealed a progressive deterioration, loss of the joint space width and changes in the subchondral bone. These alterations were accompanied by an increase of cartilage matrix degradation. Increased aggrecan and collagen type II degradation products, generated by MMPs were detected with DIAPEN and COL2-3/4C antibodies. Increased collagen type X, cellular hypertrophy and loss of proteoglycan at the articular surface were also demonstrated. A novel micro-mechanical indentation protocol revealed that the humeral heads of Fgfr3-/- mice were less stiff than those of wild type littermates. On the other hand, young adult Fgfr3-/- mice are osteopenic due to reduced cortical bone thickness and defective trabecular bone mineralization. The reduction in mineralized bone and lack of trabecular connectivity observed by micro-computed tomography were confirmed by histological and histomorphometric analyses, which revealed a significant decrease in calcein labeling of mineralizing surfaces and a significant increase in osteoid in the long bones of 4-month-old Fgfr3-/- mice. Primary cultures of adherent bone marrow-derived cells from Fgfr3-/- mice expressed markers of differentiated osteoblasts but developed fewer mineralized nodules than Fgfr3+/+ cultures of the same age. These data point to a major role for FGFR3 signaling in development and homeostatic maintenance of cartilage and bone post-natally and identify FGFR3 as a potential target for intervention in degenerative disorders of cartilage, osteopenia and those associated with defective bone mineralization.

Thesis (Ph. D.)--University of Alabama at Birmingham, 2008. / Title from first page of PDF file (viewed Feb. 13, 2009). Includes bibliographical references.

Thesis (doctoral)--Universität, München, 2001. / Includes bibliographical references (p. 117-127) and index.