O urânio (U) é um elemento tóxico radioativo encontrado na natureza, normalmente presente na água e nos alimentos e acumula-se preferencialmente em ossos. Nestes, a medula óssea constitui o alvo com o maior risco radiobiológico. Foram utilizados 60 ratos wistar recém desmamados, com vinte e dois dias de vida. Destes, trinta e cinco foram tratados com ração suplementada de 50ppm (parte por milhão) de Nitrato de Uranila e vinte e cinco foram mantidos como controle. Os animais tratados foram separados em seis grupos com cinco animais cada e os grupos controle com três animais. Foi feita a eutanásia dos 5 animais de cada grupo alimentado com urânio e 3 animais de cada grupo de controle com intervalo de tempo de 3 e 4 dias para avaliar alterações histopatológicas, hematológicas, na densidade mineral óssea e medir o teor de urânio acumulado em ossos, em função do tempo, utilizando a técnica de registro de traços de fissão SSNTD (Solid State Nuclear Track Detector). Nas avaliações histopatológicas foi observada congestão, fibrose e necrose hepática, degeneração vacuolar e desarranjo cordonal dos hepatócitos. Essas alterações iniciaram-se em animais alimentados durante três dias com ração contendo U e se intensificaram nos animais tratados durante onze dias, sugerindo que tenha ocorrido combinação de efeitos toxicológicos e radiobiológicos. Foi observada degeneração vacuolar, cilindros hialinos, fibrose e necrose nos rins dos animais alimentados com ração suplementada de U, a partir de quatorze dias de alimentação, decorrentes da nefrotoxicidade do Nitrato de Uranila. Foi observado que não ocorre alteração da densidade mineral óssea no curto prazo; porém, os animais tratados durante 21 e 28 dias, ou seja, expostos ao U por período mais longo, tiveram a densidade mineral óssea diminuída. Ocorreu substancial acúmulo de urânio nos ossos, onde foi observado 1,139 ± 0,057 ppm em ossos e 0,705 +- 0,092 ppm em dentes. Os animais dos grupos controle apresentaram teor de urânio praticamente constante no decorrer do estudo. Não foi observada alteração do teor de urânio em ração comercial. / Uranium (U) is a radioactive toxic element found in the environment, naturally present in water and food, with preference for accumulation in bone. In the latter, marrow is the target with the highest radiobiological risk. It was carried out a study with sixty Wistar rats, twenty two days old, starting at the post weaning period. From this total, thirty five animals fed with chow containing Uranyl Nitrate at a concentration of 50 ppm (parts per million) were selected as the treated group, while the remaining twenty five were the control group. Treated animals were divided into six groups with five animals each plus six control groups with three animals each. Five animals of the treated group and three of the control group were sacrificed at intervals of four days to observe histopathologic, hematologic, and bone mineral density (BMD) alterations, as well as to measure the uranium content in bone as function of time, using the Solid State Nuclear Track Detector technique. It was observed congestion, vacuolar degeneration, hepatocytes misalignment, fibrosis and necrosis in liver. These alterations were initiated in treated animals fed for three days with diets containing U and intensified in the animals treated for eleven days, suggesting the occurrence of an intertwining between radiobiological and toxicological effects. It was also observed vacuolar degeneration, hyaline cylinders, fibrosis and necrosis in the kidneys of the treated animals, all initiated after fourteen days of treatment, and these effects were attributed to the nephrotoxic character of the Uranyl Nitrate. It was found out that the BMD was not altered in the short range term of treatment, that is, treatments of twenty-one and twenty-eight days, but appreciably reduced in the long range term. There was substantial accumulation of uranium in bones and teeth, where it was measured concentrations of 1.139 ± 0.057 ppm and 0.705 ± 0.092 ppm, respectively. The uranium concentration in the bones of animals of the control group were low and approximately constant.

Este estudo investigou o efeito da Roziglitazona (RTZ) sobre a perda óssea alveolar induzida pela periodontite (POAIP). Durante 3 semanas, ratos receberam sal puro de RTZ (i.p.) ou a formulaço comercial Avandia<font face=\"Symbol\">&#210; (v.o.); os grupos controles receberam os repectiovos veículos (DMSO ou CMC). Duas semanas após o inicio do tratamento, a periodontite (P) foi induzida. Após 7 dias da indução da P, as mandíbulas foram removidas para mediço da perda óssea alveolar. Amostras de osso alveolar foram analisadas por qPCR para RUNX2, Osterix, TRAF6, TRAF2, RANKL, óxido nítrico sintases (e, n e iNOS) e PPARs (<font face=\"Symbol\">a, <font face=\"Symbol\">b e <font face=\"Symbol\">g). A farmacocinética da RTZ para cada formulaço foi estudada por HPLC-MS/MS. Tanto o sal puro como a formulaço comercial de RTZ resultou no agravamento da POAIP. Apesar dos resultados similares nas concentrações plasmáticas de RTZ os mecanismos de sinalizaço parecem depender da formulaço administrada a qual pode ser devido a interferência do veículo. / This study investigate the effects of rosiglitazone (RTZ) on periodontitis-induced alveolar bone loss (PIABL). Rats received RTZ during 3 weeks, either as the pure maleate salt (i.p.) or the commercial formulation Avandia<font face=\"Symbol\">&#226; (p.o.); control animals received the respective vehicles (DMSO or CMC). Two weeks after the treatments begins, periodontitis (P) were induced. After 7 days after P induction, jaws were removed for ABL measurement. Alveolar bone samples were analyzed by qPCR for RUNX2, Osterix, TRAF6, TRAF2, RANKL, nitric oxide sintase (e, n and iNOS) and PPARs (<font face=\"Symbol\">a, <font face=\"Symbol\">b e <font face=\"Symbol\">g). RTZ pharmacokinetics from each formulation was also studied (HPLC-MS/MS). RTZ, either from the pure maleate salt or the commercial Avandia, resulted in aggravated PIABL. Despite resulting in similar plasma RTZ concentrations, signaling mechanisms seem to depend on the administered formulation which could be due to vehicle related effects interfence.

A óxido nítrico sintase induzível (iNOS) é uma enzima responsável pela síntese do óxido nítrico, envolvido na regulação de vários processos fisiológicos, com destaque para relevantes efeitos sobre o tecido ósseo. Entretanto, o papel da iNOS no reparo ósseo alveolar permanece pouco conhecido. Assim, o objetivo deste estudo foi caracterizar o papel da iNOS no processo de reparo ósseo alveolar pósexodontia em condições homeostáticas (controle) e infecciosas em camundongos. Para isso, foram utilizados animais das linhagens WT (C57Bl/6) e iNOSKO, divididos em 2 grupos, condição homeostática (exodontia do incisivo superior direito) e alveolite experimental (induzida pela inoculação de uma suspensão de cultura bacteriana), e analisados de acordo com os diferentes períodos experimentais (0, 7, 14 e 21 dias pós-exodontia), através da análise histomorfométrica e molecular. Na análise histomorfométrica, avaliou-se a densidade de volume das estruturas referentes ao coágulo sanguíneo, tecido conjuntivo e tecido ósseo. Na análise molecular, quantificou-se a expressão de mRNA codificando genes de marcadores do metabolismo ósseo; marcadores de osteoclastos; citocinas e quimiocinas, através de reações de RealTimePCR. A expressão de iNOS esteve presente durante todo o processo de reparo ósseo alveolar nos camundongos C57Bl/6, porém de maneira mais expressiva no período de 7 dias pós-exodontia, e se mostrou aumentada pela indução de alveolite. A análise histomorfométrica demonstrou discretas alterações no processo de reparo ósseo alveolar, na ausência de iNOS, tanto em condições homeostáticas quanto infecciosas. Em condições homeostáticas, a ausência de iNOS não impactou de forma significativa o processo de reparo ósseo alveolar pósexodontia, mas se mostrou associada à modulação de vasos sanguíneos. Já em condições infecciosas, a ausência de iNOS se mostrou associada à modulação de células inflamatórias, osteoblastos e osteoclastos. De forma geral, conclui-se que embora a enzima iNOS module certos aspectos do processo de reparo ósseo alveolar de forma pontual, sua ausência não interferiu de forma significativa nesse processo. / The inducible nitric oxide synthase (iNOS) is an enzyme responsible for the synthesis of nitric oxide, a reactive molecule involved in the regulation of several physiological processes, highlighting relevant effects on bone tissue. However, the role of iNOS in alveolar bone repair remains unknown. The aim of this study was to characterize the role of iNOS in alveolar bone healing process after dental extraction in homeostatic and infectious conditions in mice. With this aim, WT (C57Bl/6) and iNOSKO mice strains were divided into 2 groups, homeostatic condition (extraction of the upper right incisor) and experimental alveolitis (induced by inoculation of a suspension of bacterial culture), and analyzed according to the different experimental periods (0, 7, 14 and 21 days post-extraction), through molecular and histomorphometric analysis. In histomorphometric analysis, the volume density of structures related to blood clot, tissue and bone were evaluated. The molecular analysis quantified the expression of mRNA encoding genes of defined as bone metabolism markers; osteoclast markers, as well cytokines and chemokines through RealTimePCR reactions. The expression of iNOS was detected during the all process of the alveolar bone repair in C57Bl/6 mice, with an expression peak at 7 days post-extraction time point, and was significantly enhanced by alveolitis induction. Histomorphometric analysis showed small changes in alveolar bone repair process in the absence of iNOS, in both homeostatic as infectious conditions. Under homeostatic conditions, the absence of iNOS did not impact significantly the process of alveolar bone healing after dental extraction, but it was associated with modulation of blood vessels formation. In the infectious conditions scenario, the absence of iNOS was associated with modulation of inflammatory cells, osteoblasts and osteoclasts counts. In general, it is concluded that even the enzyme iNOS module some aspects of alveolar bone repair in a particular and punctual manner, their absence does not interfere significantly in the overall outcome of alveolar bone repair process.

Sabe-se que o estudo da anatomia é de fundamental importância para todo e qualquer procedimento médico-cirúrgico, como também para o entendimento de toda a fisiologia e das doenças que acometem os seres vivos. Porém, para a prática da odontologia veterinária, especialidade que vem crescendo sobremaneira nos últimos anos, nota-se a ausência de uma compilação única do estudo anatômico da cabeça das espécies mais tratadas, especificamente cães e gatos. Considera-se, portanto, oportuno realizar um estudo geral dos ossos do crânio, dando ênfase ao sistema estomatognático e incluindo a articulação temporomandibular, visando descrever, ilustrar e correlacionar suas estruturas, aplicando este conhecimento na prática cirúrgica, ressaltando os pontos de maior importância para o desempenho da especialidade, disponibilizando uma base de conhecimento que atue como um \"guia&quot; para o médico-veterinário que se interessa e pratica a odontologia veterinária. Para a realização das ilustrações, crânios de cães e gatos foram preparados pela técnica de maceração, fotografados e radiografados, com suas estruturas ósseas identificadas de acordo com as necessidades da correlação cirúrgica. / The study of anatomy is important to accomplish any kind of surgical and medical procedure and to understand the physiology and the diseases that happen in animals. Nowadays, veterinary dentistry is an important area of veterinary medicine which has been increasing and improving during the last years. However, currently, the veterinarian can not find any specific and exclusive literature about head anatomy of dogs and cats. It is therefore important to perform a study of the bones of the cranium, specially the bones of the stomatognathic system, including the temporomandibular joint, describing and illustrating the most important structures and correlating this knowledge with the surgical procedures. This paper serves the veterinarians that work with dentistry as a \"guide&quot;. The illustrations and the radiographs were made with macerated craniums of dogs and cats and had the structures identified according to the surgical necessities.

Glicocorticóides são utilizados no tratamento de doenças auto-imunes e inflamatórias. Um novo composto, arilpirazola (L5) exibiu efeito antiinflamatórios e um perfil reduzido de efeitos colaterais. Avaliamos ações antiinflamatórias do L5 in vivo e os efeitos do L5 na estrutura e biomecânica ósseas em camundongos C57BL/6J. Prednisolona (Pred) e L5 reduziram o número de total leucócitos na dose de 2,1 mg/kg.pc/dia e 2,4 mg/kg.pc/dia, respectivamente. A Pred reduziu a massa corporal e o L5. Pred e L5 promoveram aumento na massa do coração. A Pred promoveu redução na massa muscular, enquanto que o L5 não teve efeito. Pred e L5 não alteraram o tecido adiposo. Na análise por microtomografia computadorizada o tratamento com L5 diminuiu BV/TV, Tb/Sp e DA, já a Pred reduziu apenas Tb/Sp. Pred e L5 não promoveram alteração no osso cortical. Pred não alterou parâmetros biomecânicos do fêmur e da tíbia e L5 reduziu energia em quebra da tíbia. Este estudo sugere que o L5 tem o mesmo potencial anti-inflamatório da Pred e que não se mostrou deletério à massa muscular. / Glucocorticoids are used to treat autoimmune and inflammatory diseases. A new compound, arilpirazola (L5) exhibited anti-inflammatory effect and reduced side effect profile. We evaluated the anti-inflammatory actions in vivo L5 and L5 on the effects of structure and biomechanical bone in mice C57BL/6J. Prednisone (Pred) and L5 reduced the overall number of leukocytes in a dose of 2.1 mg/kg.pc/day and 2.4 mg/kg.pc/day, respectively. The Pred decreased body mass and not L5. Pred and L5 caused an increase in heart mass. The Pred promoted reduction in muscle mass, while the L5 had no effect. Pred and L5 did not alter adipose tissue. The analysis by computed microtomography treatment with L5 decreased BV/TV, Tb/Sp and DA, since the only Pred reduced Tb/Sp. Pred and L5 did not promote changes in cortical bone. Pred has not altered biomechanical parameters of the femur and tibia and L5 reduced energy breaks the tibia. This study suggests that the L5 have the same potential anti-inflammatory Pred and was not deleterious to the muscle.

Chi Keung Cheung. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 219-251). / Chapter CHAPTER I --- LITERATURE REVIEW / Chapter 1 --- The structure of bone --- p.2 / Chapter 1.1. --- The cortical bone --- p.3 / Chapter 1.2. --- The cancellous bone --- p.3 / Chapter 2 --- The composition of bone --- p.3 / Chapter 2.1. --- Bone minerals --- p.4 / Chapter 2.2. --- The organic matrix --- p.4 / Chapter 2.3. --- The bone cells --- p.9 / Chapter 2.3.1. --- The osteoblast and the osteocyte --- p.9 / Chapter 2.3.2. --- The osteoclast --- p.11 / Chapter 3 --- Bone turnover - modelling and remodelling of bone --- p.13 / Chapter 3.1. --- Postulated sequence of bone remodelling --- p.14 / Chapter 4 --- Regulation of bone resorption --- p.16 / Chapter 4.1. --- Role of osteoblast and the lining cell on bone resorption --- p.17 / Chapter 5 --- Regulation of bone formation --- p.19 / Chapter 6 --- Effects of systemic hormones and local factors on bone metabolism --- p.20 / Chapter 6.1. --- Parathyroid hormone --- p.20 / Chapter 6.2. --- "1,25-dihydroxyvitamin D3" --- p.22 / Chapter 6.3. --- Calcitonin --- p.23 / Chapter 6.4. --- Prostaglandins --- p.23 / Chapter 6.5. --- Sex hormones --- p.24 / Chapter 6.6. --- Glucocorticoid --- p.26 / Chapter 6.7. --- Growth hormone --- p.27 / Chapter 6.8. --- Insulin --- p.28 / Chapter 6.9. --- Thyroid hormones --- p.29 / Chapter 6.10. --- Other systemic and local factors --- p.30 / Chapter 7 --- Indices of bone turnover --- p.34 / Chapter 8 --- Non-biochemical indices of bone metabolism --- p.34 / Chapter 8.1. --- Radionuclide bone scan --- p.34 / Chapter 8.2. --- Radiokinetic assessment --- p.35 / Chapter 8.3. --- Bone biopsy --- p.35 / Chapter 8.4. --- Bone densitometry --- p.36 / Chapter 9 --- Biochemical indices of bone metabolism --- p.37 / Chapter 10 --- Biochemical markers of bone formation --- p.38 / Chapter 10.1. --- Alkaline phosphatase --- p.38 / Chapter 10.1.1. --- Role and origin of bone alkaline phosphatase isoenzyme --- p.39 / Chapter 10.1.2. --- Measurement of bone alkaline phosphatase --- p.41 / Chapter 10.1.2.1. --- Heat inactivation --- p.42 / Chapter 10.1.2.2. --- Chemical inactivation --- p.43 / Chapter 10.1.2.3. --- Immunological methods --- p.44 / Chapter 10.1.2.4. --- High performance liquid chromatography --- p.45 / Chapter 10.1.2.5. --- Gel electrophoresis --- p.45 / Chapter 10.1.2.6. --- Isoelectric focusing --- p.47 / Chapter 10.2. --- Osteocalcin --- p.48 / Chapter 10.3. --- Osteonectin --- p.51 / Chapter 10.4. --- Matrix Gla-protein --- p.51 / Chapter 10.5. --- Other non-collagenous proteins --- p.52 / Chapter 10.6. --- Urinary Gla concentration --- p.52 / Chapter 10.7. --- Collagen peptides and extension peptides --- p.54 / Chapter 11 --- Biochemical markers of bone resorption --- p.55 / Chapter 11.1. --- Urine hydroxyproline --- p.55 / Chapter 11.2. --- Pyridinium cross-links --- p.58 / Chapter 11.3. --- Acid phosphatase --- p.60 / Chapter 11.3.1. --- Acid phosphatase isoenzymes --- p.60 / Chapter 11.3.2. --- The band 5 acid phosphatase isoenzyme genetics and characteristics --- p.62 / Chapter 11.3.3. --- Band 5 acid phosphatase as marker of osteoclastic function --- p.64 / Chapter 11.3.4. --- Measurement of osteoclastic acid phosphatase --- p.67 / Chapter 11.3.4.1. --- Specific chemical inhibitor --- p.67 / Chapter 11.3.4.2. --- Electrophoresis --- p.67 / Chapter 11.3.4.3. --- Immunological methods --- p.68 / Chapter 12 --- Problems with current biochemical markers of bone metabolism --- p.68 / Chapter 13 --- Aims of this study --- p.70 / Chapter CHAPTER II --- PURIFICATION OF TARTRATE-RESISTANT ACID PHOSPHATASE AND THE DEVELOPMENT OF AN IMMUNOASSAY FOR IT'S MEASUREMENT / Chapter 1 --- Introduction --- p.72 / Chapter 2 --- Materials and methods --- p.75 / Chapter 2.1. --- Chemicals and reagents --- p.75 / Chapter 2.1.1. --- Apparatus --- p.76 / Chapter 2.2. --- Methods --- p.77 / Chapter 2.2.1. --- Cord serum --- p.77 / Chapter 2.2.2. --- Measurement of tartrate-resistant acid phosphatase activity --- p.77 / Chapter 2.2.3. --- Measurement of protein concentration --- p.80 / Chapter 2.2.4. --- Purification of TRACP from cord plasma --- p.82 / Chapter 2.2.4.1. --- Cation-exchange column chromatography --- p.83 / Chapter 2.2.4.2. --- Gel filtration column chromatography --- p.84 / Chapter 2.2.4.3. --- Concanavalin A-affinity column chromatography --- p.85 / Chapter 2.2.4.4. --- Preparative isoelectric focusing (IEF) --- p.86 / Chapter 2.3. --- Characterisation of purified TRACP --- p.90 / Chapter 2.3.1. --- Polyacrylamide gel electrophoresis (PAGE) --- p.91 / Chapter 2.3.2. --- "Optimum pH, substrate specificity and the effects of potential activators and inhibitors on TRACP activity" --- p.99 / Chapter 2.3.3. --- Amino acid composition of purified TRACP --- p.101 / Chapter 2.4. --- Methods for raising anti-human TRACP antibody and characterisation of the antiserum --- p.102 / Chapter 2.4.1. --- Production of rabbit anti-human TRACP antibody --- p.102 / Chapter 2.4.2. --- Determination of the titre of rabbit anti-human TRACP antibody --- p.103 / Chapter 2.4.3. --- Immunoblotting analyses for cross reactivity study --- p.103 / Chapter 2.4.4. --- Immunohistochemical study for antibody specificity --- p.105 / Chapter 2.4.5. --- Cross reactivity study of the rabbit anti-human TRACP antibody to some tissue preparations --- p.107 / Chapter 2.5. --- Enzyme linked immunosorbent assay for TRACP --- p.109 / Chapter 2.5.1. --- Optimisation and evaluation of the new ELISA method for TRACP --- p.111 / Chapter 3 --- RESULTS --- p.113 / Chapter 3.1. --- "Precision of methods for the determination of protein, TRACP and phosphate." --- p.113 / Chapter 3.2. --- Isolation and purification of TRACP --- p.113 / Chapter 3.2.1. --- Concanavalin A affinity chromatography --- p.120 / Chapter 3.2.2. --- Isoelectric focusing (IEF) --- p.120 / Chapter 3.3. --- Characterisation and homogeneity of purified TRACP --- p.128 / Chapter 3.3.1. --- Characterisation of purified TRACP --- p.128 / Chapter 3.3.2. --- Homogeneity of purified TRACP --- p.132 / Chapter 3.3.3. --- Amino acid composition --- p.136 / Chapter 3.4. --- Characterisation of the rabbit anti-human TRACP antibody --- p.136 / Chapter 3.4.1. --- Antibody specificity - immunoblotting study --- p.139 / Chapter 3.4.2. --- Antibody specificity - cross reactivity with partially purified non-cord plasma TRACP --- p.142 / Chapter 3.4.3. --- Antibody specificity - immunohistochemical study --- p.145 / Chapter 3.5. --- Enzyme linked immunosorbent assay for TRACP --- p.145 / Chapter 3.5.1. --- Optimal concentration of antigen for coating of microtitre plate --- p.145 / Chapter 3.5.2. --- Kinetics of reaction with the primary rabbit anti-human TRACP antibody --- p.149 / Chapter 3.5.3. --- "Precision, recovery and assay range" --- p.149 / Chapter 4 --- DISCUSSION --- p.155 / Chapter 4.1. --- Purification of cord plasma TRACP --- p.155 / Chapter 4.2. --- Characterisation of cord plasma TRACP --- p.158 / Chapter 4.3. --- Characterisation of rabbit anti-human TRACP antibody --- p.163 / Chapter 4.4. --- Enzyme immunoassay for TRACP --- p.165 / Chapter CHAPTER III --- STUDY OF SERUM TRACP IN HEALTHY SUBJECTS AND IN PATIENTS WITH BONE RELATED DISEASES / Chapter 1 --- Introduction --- p.168 / Chapter 2 --- Materials and methods --- p.171 / Chapter 2.1. --- Subjects --- p.171 / Chapter 2.1.1. --- Healthy subjects --- p.171 / Chapter 2.1.2. --- Patients --- p.172 / Chapter 2.1.2.1. --- Post-menopausal women on hormone replacement therapy --- p.172 / Chapter 2.1.2.2. --- Hip fracture patients --- p.173 / Chapter 2.1.2.3. --- Other patients --- p.174 / Chapter 2.3. --- Measurement of other biochemical parameters --- p.175 / Chapter 2.3.1. --- Bone alkaline phosphatase --- p.175 / Chapter 2.3.2. --- "Measurement of urine hydroxyproline, creatinine, calcium, osteocalcin, thyroid hormones and parathyroid hormone" --- p.176 / Chapter 2.4. --- Statistics --- p.178 / Chapter 3 --- RESULTS --- p.179 / Chapter 3.1. --- Healthy subjects --- p.179 / Chapter 3.2. --- Serum TRACP concentration in post-menopausal women before and after hormone replacement therapy --- p.185 / Chapter 3.3. --- TRACP concentration in elderly subjects with hip fractures --- p.189 / Chapter 3.4. --- Serum TRACP concentrations in patients with other bone related diseases --- p.190 / Chapter 3.4.1. --- Hyperthyroidism --- p.194 / Chapter 3.4.2. --- Hyperparathyroidism --- p.198 / Chapter 3.4.3. --- Haemodialysis --- p.201 / Chapter 4 --- DISCUSSION --- p.204 / GENERAL DISCUSSION --- p.216 / REFERENCES --- p.219

骨科醫生面臨著老年婦女的骨修復受損或者癒合延遲的挑戰，這使得康復過程變長，甚至引發高死亡率。至今為止，臨床上仍然沒有促進老年骨癒合的滿意治療方法，因此亟需其他治療策略。骨癒合重現了胚胎後的骨骼發育過程。直接由骨外膜成骨（膜內骨化）以及通過軟骨介質成骨（軟骨內骨化）是骨癒合中的兩個重要過程。 雌激素受體β（ERβ基因敲除雌性小鼠的研究表明ERβ信號通路在骨骼發育過程中同時參與了抑制膜內骨化和軟骨內骨化這兩個過程。臨床活檢的資料顯示，在絶經後婦女的骨痂中，ERβ陽性的增生軟骨細胞數量增加。然而，ERβ在正常和老年骨癒合的作用還沒有研究。 / 本研究通過下述部分檢查了ERβ在正常和老年骨癒合的作用，以及其將來的藥物應用：1) 建立一個以膜內骨化為主的骨癒合模型。2) 通過連個骨癒合模型，檢查ERβ在正常骨癒合中的作用。3) 檢查ERβ在老年骨癒合中的作用並檢查ERβ拮抗劑PHTPP 對老年骨癒合的潛在藥物療效。 / 實驗1是建立一個以膜內骨化為主的骨癒合模型。以前建立的小鼠股骨中段骨折模型是軟骨內骨化為主的骨癒合模型。由於技術難度，該模型可重複性不高，而且其金屬內固定器會造成金屬偽影，進而不能應用高解析度微焦點CT跟蹤觀察的技術。為了檢查ERβ在膜內骨化中的作用，並且應用微焦點CT跟蹤觀察技術，我們首先建立了一個小鼠鑽孔缺損模型。該實驗同時也確認了去勢誘導的骨質疏鬆小鼠相比正常小鼠，在鑽孔缺損模型中骨癒合受阻。 / 實驗2檢驗了阻斷ERβ能促進正常骨癒合的假設。本實驗應用ERβ基因敲除小鼠，在兩個模型中檢驗了實驗假設。第一個是傳統的小鼠股骨中段骨折模型，第二個是由實驗1建立的鑽孔缺損模型。兩個模型都證實ERβ基因敲除小鼠骨癒合和野生型小鼠相比，早期的血管新生和中期的礦化有所增強，末期的骨癒合沒有明顯差異。 / 實驗3 進一步研究ERβ在老年骨癒合中的作用。實驗應用老年小鼠股骨中段骨折模型，比較ERβ基因敲除小鼠和野生型小鼠之間的癒合過程。結果顯示ERβ基因敲除小鼠骨癒合和野生型小鼠相比，早期的血管新生，中期的礦化以及末期的力學性能都有所增強。該結果預示阻斷ERβ能作為另一種治療老年骨折癒合的治療策略。同時，我們也檢測了ERβ的拮抗劑PHTPP（4 - [2 - 苯基- 5,7 -二（三氟甲基）吡唑並[1,5 - A]嘧啶3 - 基]苯酚, 在老年骨癒合中的治療效果。 通過比較用藥組小鼠與安慰劑組小鼠的骨癒合品質，顯示PHTPP治療小鼠血管新生，骨痂礦化和最終的力學性質均優於對照安慰劑組小鼠。 / 綜上所述，本研究描述了ERβ在正常和老年骨癒合中的作用。骨癒合的關鍵過程包括血管新生，膜內骨化以及軟骨內骨化在阻斷ERβ後都得到增強，從而加快正常骨和老年骨的骨痂形成，礦化並增強力學性質。ERβ的拮抗劑PHTPP在老年小鼠骨折模型中能促進骨癒合。本研究提出了一個新的骨癒合治療策略，並為將來的臨床實驗提供了堅實的基礎。 / Orthopaedic surgeons are challenged by impaired or delayed bone healing in elderly women, which requires prolongation of rehabilitation process or even induces high mortality. Up to date, there are no satisfactory therapeutic modalities for promoting aged bone healing clinically, and alternative therapeutic stratagem is therefore desirable. Bone healing recapitulates postnatal bone development. Direct periosteam-dependent bone formation (intramembranous ossification) and the formation of bone through a cartilage intermediate (endochondral ossification) are the two important processes during bone healing. Evidences from Estrogen Receptor β (ERβ), gene knockout female mouse studies have demonstrated that ERβ signaling participates in inhibiting both intramembranous and endochondral ossification during bone development. Clinical biopsy data demonstrated that the number of ERβ positive proliferative chondrocytes within fracture callus was increased in postmenopausal women. However, the role of ERβ in normal and aged bone healing is not examined yet. / This study examined role of ERβ in normal and aged bone healing and the future pharmaceutical application though the following part: 1) Establish an intramembranous ossification-dominant bone healing model. 2) Examine the role of ERβ in normal bone healing though two models. 3) Examine the role of ERβ in aged bone healing and investigate the potential therapeutical efficacy of an ERβ antagonist PHTPP in aged bone healing. / Study I was to establish an intramembranous ossification dominant bone healing mouse model. Previous available mouse femoral shaft fracture model was a endochondral ossification dominant bone healing model. This model was technically difficult to generate high reproducibility and the inside metal stabilization devices prevented the application of high-resolution in vivo micro-CT monitoring due to the metal artifact. In order to examine the role of ERβ in intramembranous ossification and apply the micro-CT monitoring technique, a drill-hole defect mouse model was developed. The study also confirmed bone healing was impaired in mice with ovariectomy -induced osteoporosis in drill-hole defect model. / Study II was to test the hypothesis that blockade of ERβ could promote normal bone healing. ERβ knockout mice were employed in this study and the hypothesis was examined in two models, the first is the traditional mouse femoral shaft fracture model, and the second is the drill-hole defect model that was developed in study I. Both models demonstrated that the bone healing in ERβ knockout mice was enhanced in the early stage of neovascularization and the middle stage of ossification but not by the end of healing compare to the wild type mice. / Study III was designed to further investigate the role of ERβ in aged bone healing. Femoral shaft fracture model was created in aged mice. The healing process was compared between the ERβ knockout mice and wild type mice. The results demonstrated that ERβ knockout mice was enhanced in the early stage of neovascularization, the middle stage of ossification and end stage of mechanical strength. The findings implied blockade of ERβ can be considered as another therapeutic strategy for aged fracture healing. PHTPP (4-[2-Phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl] phenol), an ERβ antagonist, was employed in aged mice femoral shaft fracture model. The bone healing quality of treated mice was compared with that of the vehicle control mice. It showed PHTPP treated mice had enhanced neovascularization, callus ossification and finally better mechanical properties than vehicle mice. / The present study depicted the role of ERβ in normal and aged bone healing. Key processes including neovascularization, intramembranous and endochondral ossification were all enhanced by blockade of ERβ, which led to fast callus formation, mineralization in normal bone and better mechanical properties in aged bone. ERβ antagonist PHTPP could promote aged bone healing in mouse osteotomy model. This study raised an alternative therapeutic stratagem for bone healing and provided solid basis for future clinical trials. / 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. / He, Yixin. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 147-167). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / ABSTRACT --- p.i / 中文摘要 --- p.iv / PUBLICATIONS AND AWARDS --- p.vi / ACKNOWLEDGEMENTS --- p.xi / TABLE OF CONTENTS --- p.xii / LIST OF ABBREVIATIONS --- p.xvi / LIST OF FIGURES --- p.xviii / LIST OF TABLES --- p.xx / Chapter CHAPTER 1 --- INTRODUCTION AND LITERATURE REVIEW --- p.1 / Chapter 1.1 --- Fracture and Bone Healing --- p.2 / Chapter 1.1.1 --- Epidemiology and Impacts of Fractures --- p.2 / Chapter 1.1.2 --- Current Management and Limitations --- p.3 / Chapter 1.1.3 --- Bone Structures --- p.5 / Chapter 1.1.4 --- Bone Healing --- p.7 / Chapter 1.1.5 --- Aged Bone Healing --- p.12 / Chapter 1.1.6 --- Enhancements of Bone Healing --- p.17 / Chapter 1.2 --- Estrogen and Estrogen Receptors --- p.19 / Chapter 1.2.1 --- Estrogen Receptors α and β --- p.19 / Chapter 1.2.2 --- Molecular Actions of Estrogens --- p.20 / Chapter 1.2.3 --- Estrogen receptors in bone homeostasis --- p.24 / Chapter 1.3 --- Hypothesis --- p.28 / Chapter 1.4 --- Study Plan and Objectives --- p.32 / Chapter 1.4.1 --- Bone Healing Models --- p.32 / Chapter 1.4.2 --- Study Outline --- p.32 / Chapter 1.5 --- Figures and Tables --- p.34 / Chapter CHAPTER 2 --- ESTABLISHMENT OF DRILL-HOLE DEFECT HEALING MODEL IN MICE --- p.39 / Chapter 2.1 --- Introduction --- p.40 / Chapter 2.1.1 --- Limitations in currently available mouse models of osteoporotic bone healing --- p.40 / Chapter 2.1.2 --- Creation of a drill-hole defect at the mid-diaphysis of the femur for in vivo monitoring of bone healing in mice --- p.40 / Chapter 2.2 --- Materials and Methods --- p.43 / Chapter 2.2.1 --- Experimental animals --- p.43 / Chapter 2.2.2 --- Surgical protocol and experimental design --- p.43 / Chapter 2.2.3 --- Micro-CT analysis of intact femur --- p.44 / Chapter 2.2.4 --- In vivo micro-CT analysis of new bone formation in the drill-hole site --- p.45 / Chapter 2.2.5 --- Micro-CT-based angiography --- p.45 / Chapter 2.2.6 --- Histological examination --- p.46 / Chapter 2.2.7 --- Immunohistochemistry --- p.46 / Chapter 2.2.8 --- Quantitative real-time PCR --- p.47 / Chapter 2.2.9 --- Analysis of bone formation and resorption markers --- p.47 / Chapter 2.2.10 --- Mechanical testing --- p.48 / Chapter 2.2.11 --- Statistical analysis --- p.48 / Chapter 2.3 --- Results --- p.51 / Chapter 2.3.1 --- Confirmation of osteoporotic bone prior to generation of a drill-hole defect --- p.51 / Chapter 2.3.2 --- General observation of mice following drill-hole surgery --- p.51 / Chapter 2.3.3 --- In vivo micro-CT analysis of new bone in the drill-hole site of mouse femurs --- p.51 / Chapter 2.3.4 --- In vivo micro-CT analysis of new bone in drill-hole sites is highly reproducible --- p.52 / Chapter 2.3.5 --- Micro-CT angiography --- p.52 / Chapter 2.3.6 --- Histological observation of bone healing --- p.53 / Chapter 2.3.7 --- Immunohistochemical analysis of ER expressions during bone healing --- p.54 / Chapter 2.3.8 --- Quantitative real-time PCR analysis of gene expression during bone healing --- p.54 / Chapter 2.3.9 --- Analysis of bone formation and resorption markers during bone healing --- p.54 / Chapter 2.3.10 --- Mechanical testing of femurs from Sham and OVX mice --- p.55 / Chapter 2.4 --- Discussion --- p.56 / Chapter 2.4.1 --- Bone healing with dominant intramembranous ossification --- p.56 / Chapter 2.4.2 --- Impaired osteoporotic bone healing --- p.57 / Chapter 2.4.3 --- Reproducibility of the in vivo micro-CT method for analysis of bone healing --- p.58 / Chapter 2.4.4 --- Dysregulated expression of estrogen receptors and bone healing in OVX mice --- p.59 / Chapter 2.4.5 --- Study limitations --- p.60 / Chapter 2.4.6 --- Conclusions --- p.60 / Chapter 2.5 --- Figures and Tables --- p.61 / Chapter CHAPTER 3 --- ROLE OF ERβ IN NORMAL BONE HEALING --- p.72 / Chapter 3.1 --- Introduction --- p.73 / Chapter 3.2 --- Materials and Methods --- p.75 / Chapter 3.2.1 --- Part I Study --- p.75 / Chapter 3.2.1.1 --- Experimental animals --- p.75 / Chapter 3.2.1.2 --- Fracture model and experimental design --- p.75 / Chapter 3.2.1.3 --- Radiographic Analysis --- p.76 / Chapter 3.2.1.4 --- Micro-CT-based angiography --- p.76 / Chapter 3.2.1.5 --- Micro-CT analysis of callus --- p.77 / Chapter 3.2.1.6 --- Histological examination --- p.78 / Chapter 3.2.1.7 --- Dynamic Bone histomorphometric analysis --- p.78 / Chapter 3.2.1.8 --- Mechanical testing --- p.79 / Chapter 3.2.1.9 --- Quantitative real-time PCR --- p.80 / Chapter 3.2.1.10 --- Analysis of bone formation and resorption markers --- p.80 / Chapter 3.2.1.11 --- Statistical analysis --- p.81 / Chapter 3.2.2 --- Part II Study --- p.81 / Chapter 3.2.2.1 --- Experimental animals and design --- p.81 / Chapter 3.2.2.2 --- Evaluation protocols --- p.82 / Chapter 3.2.2.3 --- Statistical analysis --- p.82 / Chapter 3.3 --- Results --- p.83 / Chapter 3.3.1 --- Part I Study --- p.83 / Chapter 3.3.1.1 --- Radiographic Analysis --- p.83 / Chapter 3.3.1.2 --- Micro-CT angiography --- p.83 / Chapter 3.3.1.3 --- Micro-CT analysis of callus --- p.83 / Chapter 3.3.1.4 --- Histological and dynamic histomorphometric analysis --- p.84 / Chapter 3.3.1.5 --- Mechanical testing of the callus --- p.85 / Chapter 3.3.1.6 --- Quantitative real-time PCR analysis of gene expression --- p.85 / Chapter 3.3.1.7 --- Analysis of bone formation and resorption markers during bone healing --- p.85 / Chapter 3.3.2 --- Part II Study --- p.86 / Chapter 3.3.2.1 --- In vivo micro-CT analysis of new bone in the drill-hole site of mouse femurs --- p.86 / Chapter 3.3.2.2 --- Micro-CT angiography --- p.87 / Chapter 3.3.2.3 --- Histological observation of bone healing --- p.87 / Chapter 3.3.2.4 --- Quantitative real-time PCR analysis of gene expression --- p.88 / Chapter 3.3.2.5 --- Analysis of bone formation and resorption markers during bone healing --- p.88 / Chapter 3.3.2.6 --- Mechanical testing of femurs from WT and KO mice --- p.88 / Chapter 3.4 --- Discussion --- p.90 / Chapter 3.4.1 --- Angiogenesis --- p.90 / Chapter 3.4.2 --- Fracture Healing --- p.91 / Chapter 3.4.3 --- Estrogen receptor β and endochondral and intramembranous ossification --- p.93 / Chapter 3.4.4 --- Estrogen receptor β in aged bone --- p.94 / Chapter 3.4.5 --- Conclusions --- p.94 / Chapter 3.5 --- Figures and Tables --- p.95 / Chapter CHAPTER 4 --- ROLE OF ERβ AND ITS ANTAGONIST PHTPP IN AGED BONE HEALING --- p.113 / Chapter 4.1 --- Introduction --- p.114 / Chapter 4.2 --- Materials and Methods --- p.116 / Chapter 4.2.1 --- Experimental animals --- p.116 / Chapter 4.2.2 --- Fracture model and experimental design --- p.116 / Chapter 4.2.3 --- Radiographic Analysis --- p.117 / Chapter 4.2.4 --- Micro-CT-based angiography --- p.118 / Chapter 4.2.5 --- Micro-CT analysis of callus --- p.118 / Chapter 4.2.6 --- Histological examination --- p.119 / Chapter 4.2.7 --- Dynamic Bone histomorphometric analysis --- p.120 / Chapter 4.2.8 --- Mechanical testing --- p.120 / Chapter 4.2.9 --- Quantitative real-time PCR --- p.121 / Chapter 4.2.10 --- Analysis of bone formation and resorption markers --- p.122 / Chapter 4.2.11 --- Statistical analysis --- p.122 / Chapter 4.3 --- Results --- p.123 / Chapter 4.3.1 --- Radiographic Analysis --- p.123 / Chapter 4.3.2 --- Micro-CT angiography --- p.123 / Chapter 4.3.3 --- Micro-CT analysis of callus --- p.123 / Chapter 4.3.4 --- Histological and dynamic histomorphometric analysis --- p.124 / Chapter 4.3.5 --- Mechanical testing of the callus --- p.125 / Chapter 4.3.6 --- Quantitative real-time PCR analysis of gene expression during fracture healing --- p.125 / Chapter 4.3.7 --- Analysis of bone formation and resorption markers during bone healing --- p.126 / Chapter 4.4 --- Discussion --- p.127 / Chapter 4.4.1 --- Angiogenesis --- p.127 / Chapter 4.4.2 --- Fracture Healing --- p.128 / Chapter 4.4.3 --- Estrogen receptor β and endochondral ossification --- p.129 / Chapter 4.4.4 --- ERβ antagonist PHTPP --- p.130 / Chapter 4.4.5 --- Conclusions --- p.130 / Chapter 4.5 --- Figures and Tables --- p.131 / Chapter CHAPTER 5 --- STUDY LINITATIONS, FURTHER RESEARCH AND CONCLUDSIONS --- p.142 / Chapter 5.1 --- Limitations --- p.143 / Chapter 5.1.1 --- Bone healing model --- p.143 / Chapter 5.1.2 --- Estrogen receptors and transgenic mouse --- p.143 / Chapter 5.1.3 --- ERβ antagonist PHTPP --- p.144 / Chapter 5.2 --- Further Research --- p.144 / Chapter 5.2.1 --- ERβ signaling --- p.144 / Chapter 5.2.2 --- Preclinical Trial --- p.145 / Chapter 5.3 --- Conclusions --- p.146 / BIBLIOGRAPHY --- p.147

by Tang Shengping. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (p. 217-244). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Yim, Po Yee Annie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 166-219). / Abstracts in English and Chinese. / Acknowledgements --- p.ii / Abstract --- p.iv / Abbreviations --- p.xi / Table of Content --- p.xiii / List of Figures --- p.xviii / List of Tables --- p.xxi / Major Conference Presentations --- p.xxiii / Publication in Preparation --- p.xxvi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- General Overview of Adolescent Idiopathic Scoliosis (AIS) --- p.2 / Chapter 1.2 --- Natural History --- p.3 / Chapter 1.3 --- Current Treatments --- p.5 / Chapter 1.3.1 --- Non-operative Treatments --- p.5 / Chapter 1.3.2 --- Surgical Treatments --- p.6 / Chapter 1.4 --- Current Hypothesis on the Etiology of AIS --- p.8 / Chapter 1.4.1 --- Genetic Factors --- p.8 / Chapter 1.4.2 --- Neuromuscular Impairment --- p.10 / Chapter 1.4.3 --- Abnormalities in Skeletal Development --- p.11 / Chapter 1.4.4 --- Metabolic Dysfunction --- p.12 / Chapter 1.4.4.1 --- Lower Bone Mineral Density --- p.12 / Chapter 1.4.4.2 --- Delayed Sexual Maturity --- p.14 / Chapter 1.4.4.3 --- Hormonal Dysfunction --- p.14 / Chapter 1.5 --- Skeletal arid Spinal Growth in AIS --- p.16 / Chapter 1.5.1 --- Abnormal Growth during Puberty --- p.16 / Chapter 1.5.2 --- Growth Pattern --- p.17 / Chapter 1.5.3 --- Disproportional Growth in AIS --- p.18 / Chapter 1.5.4 --- Asymmetric Growth --- p.20 / Chapter 1.6 --- Melatonin and its Receptor --- p.22 / Chapter 1.6.1 --- Introduction --- p.22 / Chapter 1.6.2 --- Melatonin Receptor --- p.24 / Chapter 1.6.3 --- Melatonin's Role in t h e Skeletal System --- p.25 / Chapter 1.6.4 --- Melatonin-Deficient Scoliotic Animal Model --- p.27 / Chapter 1.6.5 --- Melatonin and AIS --- p.29 / Chapter 1.6.5.1 --- Melatonin Level in AIS --- p.30 / Chapter 1.6.5.2 --- Melatonin Receptor in AIS --- p.30 / Chapter Chapter 2 --- Hypothesis and Objectives --- p.39 / Chapter 2.1 --- Study Hypothesis --- p.40 / Chapter 2.2 --- Objectives --- p.41 / Chapter Chapter 3 --- Abnormal skeletal growth patterns in adolescent idiopathic scoliosis - A longitudinal study till skeletal maturity --- p.42 / Chapter 3.1 --- Introduction --- p.43 / Chapter 3.2 --- Methodology --- p.44 / Chapter 3.2.1 --- Recruitments of Subjects --- p.44 / Chapter 3.2.1.1 --- Patients with AIS --- p.44 / Chapter 3.2.1.2 --- Normal Controls --- p.44 / Chapter 3.2.1.3 --- Patients Consents --- p.45 / Chapter 3.2.2 --- Anthropometric Measurements --- p.45 / Chapter 3.2.3 --- Data Analysis --- p.46 / Chapter 3.2.3.1 --- Cross-sectional Study --- p.46 / Chapter 3.2.3.2 --- Longitudinal Study --- p.46 / Chapter 3.3 --- Results --- p.47 / Chapter 3.3.1 --- Cross-sectional Study of Anthropometric Measurements --- p.47 / Chapter 3.3.2 --- Longitudinal Study of Anthropometric Measurements --- p.48 / Chapter 3.3.2.1 --- Comparison Adjusted for Chronological Age --- p.49 / Chapter 3.3.2.2 --- Comparison Along Year Since Menarche (YSM) --- p.49 / Chapter 3.4 --- Discussion --- p.51 / Chapter Chapter 4 --- Establishment of a Melatonin-Deficierit Induced Scoliotic Model with Locally Bred Chicken --- p.63 / Chapter 4.1 --- Introduction --- p.64 / Chapter 4.2 --- Methodology --- p.67 / Chapter 4.2.1 --- Animals --- p.67 / Chapter 4.2.2 --- Materials and Reagents --- p.67 / Chapter 4.2.3 --- Pinealectomy --- p.68 / Chapter 4.2.4 --- Confirmation of Pineal Gland Removal --- p.69 / Chapter 4.2.5 --- Development of Scoliosis --- p.69 / Chapter 4.2.6 --- Measurement of Long Bone Growth --- p.70 / Chapter 4.2.7 --- Measurement of Weight --- p.71 / Chapter 4.2.8 --- Measurement of Bone Mineral Density (BMD) --- p.71 / Chapter 4.2.8.1 --- Micro Computed Tomography (MicroCT) --- p.71 / Chapter 4.2.8.2 --- Image Processing and Evaluation of BMD --- p.71 / Chapter 4.2.9 --- Data Analysis --- p.72 / Chapter 4.2.9.1 --- Measurements of Long Bone Growth and Weight --- p.72 / Chapter 4.2.9.2 --- Bone Mineral Density --- p.72 / Chapter 4.3 --- Results --- p.73 / Chapter 4.3.1 --- Confirmation of Pineal Gland Removal --- p.73 / Chapter 4.3.2 --- Occurrence of Scoliosis --- p.73 / Chapter 4.3.3 --- Measurements of Long Bone and Weight --- p.74 / Chapter 4.3.4 --- Measurement of Bone Mineral Density --- p.75 / Chapter 4.4 --- Discussion --- p.76 / Chapter Chapter 5 --- Expression of Melatonin Receptor in AIS and Control --- p.102 / Chapter 5.1 --- Introduction --- p.103 / Chapter 5.2 --- Methodology --- p.105 / Chapter 5.2.1 --- Subjects Recruitments --- p.105 / Chapter 5.2.2 --- Cell Isolation --- p.106 / Chapter 5.2.2.1 --- Bone Biopsies for Osteoblasts Isolation --- p.106 / Chapter 5.2.2.2 --- Materials and Reagents --- p.106 / Chapter 5.2.2.3 --- Isolation of Osteoblasts from Bone Biopsies --- p.107 / Chapter 5.2.3 --- Expression Level and Pattern of Melatonin Receptors 1A and IB --- p.108 / Chapter 5.2.3.1 --- Materials and Reagents --- p.108 / Chapter 5.2.3.2 --- Validation of Specificities of Antibodies by Co-immunoprecipitation --- p.113 / Chapter 5.2.3.3 --- Quantification of Protein Expression of Melatonin Receptors in Osteoblasts --- p.115 / Chapter 5.2.3.4 --- Quantification of mRNA Expression of Melatonin Receptor in Osteoblast --- p.117 / Chapter 5.2.3.5 --- Localization of Melatonin Receptor 1A and IB by Immunofluorescence Staining --- p.119 / Chapter 5.2.4 --- Evaluation and Correlation of Clinical Phenotypes with Melatonin Receptor Expression --- p.120 / Chapter 5.2.5 --- Data Analysis --- p.120 / Chapter 5.3 --- Results --- p.121 / Chapter 5.3.1 --- Protein Expression of Melatonin Receptor 1A and IB --- p.121 / Chapter 5.3.2 --- mRNA Expression of Melatonin Receptor 1A and IB --- p.121 / Chapter 5.3.3 --- Localization of Melatonin Receptors 1A and IB --- p.122 / Chapter 5.3.4 --- Evaluation and Correlation of Clinical Phenotypes with Melatonin Receptor Expression --- p.123 / Chapter 5.4 --- Discussion --- p.124 / Chapter Chapter 6 --- Summary and Overall Discussion --- p.152 / Chapter 6.1 --- Study Flowchart --- p.153 / Chapter 6.2 --- Summary and Discussion --- p.159 / Chapter 6.3 --- Limitations and Further Studies --- p.163 / Bibliography --- p.166

Made available in DSpace on 2014-06-11T19:31:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-12-20Bitstream added on 2014-06-13T21:02:11Z : No. of bitstreams: 1 tanaka_fy_dr_araca.pdf: 1088210 bytes, checksum: 03e8b8b712b01bee40429585b8ec19b1 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O objetivo deste trabalho foi analisar a viabilidade celular comparando os meios de conservação para enxerto ósseo. A preservação de células viáveis em procedimentos de enxerto ósseo é de fundamental importância para que se tenha a osteogênese. Foram utilizados 43 ratos machos. Após a antissepsia do campo operatório foi realizada incisão linear na região mediana da calota craniana para obtenção do enxerto da região parietal direita e esquerda as quais foram removidas com auxílio de trefina de 5mm de diâmetro acoplada em micro-motor de baixa rotação, sob constante irrigação com solução de soro fisiológico 0,9% estéril. As peças do enxerto foram acondicionadas em tubos de ensaio estéreis os quais foram devidamente identificadas de acordo com o grupo e mantidas dentro deste tubo conforme cada condição do grupo. Como meio de conservação da viabilidade celular do enxerto foi utilizado o soro fisiológico a 0,9% (Grupo I) e a solução de Euro Collins® (Grupo II) e ainda para verificar se a temperatura tem influência direta na manutenção da viabilidade celular foi analisado o enxerto ósseo conservado em temperatura ambiente (Grupo III) e o enxerto ósseo sem nenhuma solução, porém mantido em gelo (GrupoIV). Para avaliar a viabilidade celular foi utilizada análise histológica e imunoistoquímica imediata e ainda em cada grupo analisou-se a viabilidade celular no período de 6 horas, 12 horas, 24 horas e 30 horas. Como resultado observou-se que a solução de Euro Collins® apresentou-se superior ao soro fisiológico no que se diz respeito à manutenção da viabilidade celular do enxerto ósseo onde se pode notar viabilidade celular até o período de 30 horas. / The aim of this study was to analyze cellular viability comparing storage media for skull vault bone graft. Preservation of viable cells in bone graft procedures is of paramount importance to obtain osteogenesis. Forty-three male used in this study. After antisepsis of the operative field, a linear incision was made on the middle region of the skull vault to obtain a bone graft from the right and left parietal areas. The grafts were removed with a 5-mm diameter trephine bur coupled to low-speed handpiece under continuous irrigation with sterile 0.9% saline. The graft pieces were placed in sterile 5-mL test tubes with caps, and were properly identified according to the group and maintained inside the test tubes as per each group conditions. The storage media evaluated for preservation of graft cellular viability were 0.9% saline (Group I) and Euro Collins® solution (Group II). In order to assess whether the temperature has a direct influence on the maintenance of cellular viability, the analysis was extended to bone grafts stored at room temperature (Group III) and bone grafts with no solution, but maintained in ice (Group IV). Cellular viability was evaluated by immediate histological and immunohistochemical analyses. For each group, cellular viability was analyzed at 6, 12, 24 and 30 hours after procedure. The results of this study showed that Euro Collins® solution yielded better performance than 0.9% saline as regards the maintenance of bone graft cellular viability (up to 30 hours).