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Investigation of activated remodelling in the healing of experimental stress fractures and the influence of anti-inflammatory treatmentsLisa Kidd Unknown Date (has links)
Investigation of activated remodelling in the healing of experimental stress fractures and the influence of anti-inflammatory treatments Lisa Jane Kidd Abstract Targeted and focal remodelling are important processes in bone homeostasis and pathology. However, the factors that initiate and direct remodelling to repair microcracks, or respond to excess loading are still poorly understood. The rat ulna-loading (RUL) model has been widely used to examine modelling and remodelling responses to axial cyclic loading. However the model has not yet been fully characterised. Stress fractures are common amongst athletes, dancers and military recruits, but there is almost no information available on the mechanism of healing of these fractures. Although cyclooxygenase-2 (COX-2) is a key mediator of bone resorption and bone formation, very little information is available on the effect of non-steroidal antiinflammatories (NSAIDs) on stress fracture healing. Remodelling may play a role in the pathogenesis of stress fractures, and there is growing interest in the potential use of bisphosphonates to prevent them. Nonetheless, the effect of bisphosphonates on stress fracture healing is not known. PMX53 is a C5a receptor antagonist developed as a novel anti-inflammatory agent. It is effective against inflammatory arthritis, but has not been tested in any fracture models. The aims of this study were to undertake a detailed examination of the histology, histomorphometry and gene expression of the healing and remodelling process initiated by RUL, and to use this model to determine the effects of selective and non-selective NSAIDs, a bisphosphonate and PMX53 on stress fracture healing. To characterise the RUL model, fatigue fractures were created by loading ulnae until displacement was observed to increase by between 4% and 50%. Ulnae were bulk-stained in basic fuchsin and processed for undecalcified histology. For all remaining experiments, loading was stopped when the displacement had increased by 10%. For detailed histology and histomorphometry, ulnae were decalcified, paraffin embedded and stained with toluidine blue, saffranin-O or for tartrate resistant acid phosphatase (TRAP). Ulnae were examined at 1, 2, 4, 6, 8, and 10 weeks after loading. The effects of DFU (a selective COX-2 inhibitor, 2 mg/kg po), ibuprofen (a non-selective NSAID, 30 mg/kg po) and PMX53 (10 mg/kg po) were examined at 2, 4 and 6 weeks after loading. Effects of risedronate (a bisphosphonate) were examined at a high (1.0 mg/kg po) and low dose (0.1 mg/kg po) at 2, 6 and 10 weeks after loading. RUL did not create isolated intracortical microcracks, but curvilinear fatigue fractures that occurred at a standard position in the medial cortex of the distal ulna diaphysis. These stress fractures induced rapid periosteal woven bone formation and direct intracortical remodelling along the fracture line that originated at the periosteum and progressed towards the medullary cavity. Basic multicellular units (BMUs) could be followed through serial sections extending along the fracture line towards the centre of the bone. Quantitative, real-time PCR was performed at 4 hours, 24 hours, 4 days, 7 days and 14 days after fatigue fracture. Following each period, bones were dissected and mRNA was extracted using standard protocols. Gene expression was compared between loaded and unloaded ulnae and to an unloaded control group. Four hours after loading, there was a marked, 220-fold increase (P<0.0001) in expression of Interleukin-6 (IL-6). There were also prominent peak increases in mRNA expression for Osteoprotegerin (OPG), cyclooxygenase-2 (COX-2), and vascular endothelial growth factor (VEGF) (all P<0.0001). At 24 hours there was a peak increase in mRNA expression for IL-11 (73-fold increase, P<0.0001). At 4 days there was a significant increase in mRNA expression for Bcl-2, COX-1, bone morphogenic protein (BMP)-2, insulin-like growth factor (IGF)-1, osteopontin (OPN), and stromal cell derived factor SDF-1. At 7 days there was a significant increase in mRNA expression of Receptor activator of nuclear factor kappa β ligand (RANKL) and OPN. The dramatic, early up-regulation of IL-6 and IL-11 suggests they play a central role in initiating signalling events for stress fracture healing. Treatment with PMX53 did not affect any measures of woven bone formation or stress fracture remodelling. There were no treatment effects of Ibuprofen or DFU on the area of woven bone. DFU treatment resulted in a significant reduction in the area of porosity (resorption) and BMU area along the fracture line at 2 weeks after fracture. Ibuprofen treatment resulted in a significant reduction in length and area of BMUs and new bone formation along the fracture line at 6 weeks (p < 0.05). This is the first report to demonstrate a negative effect on stress fracture healing of both a selective COX-2 inhibitor and a non-selective NSAID. These data confirm the importance of cyclooxygenase in bone resorption and formation during remodelling. Bisphosphonates are potent inhibitors of osteoclastic bone resorption Two, 6 and 10 weeks after loading, measures of resorption and new bone formation were significantly reduced along the fracture line by high dose risedronate treatment, but not by the low dose. Only the porosity along the fracture line 2 weeks after loading was significantly reduced by the low dose risedronate. The low dose more closely resembles the clinical dose used to treat patients. Woven bone formation and consolidation were not affected by the low or high doses of risedronate. In conclusion, fatigue fractures in the rat ulna are highly reproducible, induce exuberant periosteal woven bone formation, and heal by direct remodelling along the fracture line. Remodelling is associated with gene expression for molecules typically associated with bone resorption and formation, angiogenesis and cell signalling. Remodelling of the stress fracture line was adversely affected by treatment with selective and non-selective COX inhibitors, by high dose treatment with risedronate, but not by PMX53, a C5a antagonist.
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Approche micromécanique du remodelage osseux / Micromechanical approach of the cortical bone remodelingDevulder, Anne 29 June 2009 (has links)
Dans le cadre de la prédiction du risque fracturaire associée à diverses pathologies, comme l'ostéoporose, cette étude vise à une meilleure compréhension du comportement mécanique de l'os cortical humain, notamment à l'échelle de la microstructure, et, en particulier, du processus biologique de remodelage osseux. Ce phénomène permet, en effet, le renouvellement continuel de la microstructure au cours du temps et contribue ainsi à une diminution de l'endommagement de l'os et, par conséquent, des risques de fracture. Les facteurs déterminants et les conséquences sur les champs mécaniques locaux au sein de la microstructure sont ici recherchés. Une approche couplée, expérimentale et numérique, est proposée. Huit spécimens de fémurs humains, de sexes féminins, âgés de 74 à 101 ans sont analysés. L'analyse expérimentale est réalisée à différentes échelles. A l'échelle macroscopique, le module de Young et les paramètres à la rupture sont déterminés via des essais de compression et les relations potentielles avec les caractéristiques morphométriques, que sont l'âge, la porosité et la densité minérale, sont évaluées. L'analyse de l'évolution des champs de déformations locaux au cours de ces essais de compression et des essais de nanoindentation permet d'accéder à des échelles plus fines (micro- et nanoscopique) afin d'apprécier l'hétérogénéité de la microstructure. On s'intéresse plus particulièrement à l'endommagement de l'os et à l'étape d'initiation de microfissures ainsi qu'à l'hétérogénéité du module de Young. Macroscopiquement, le paramètre le plus influent semble être la porosité. Microscopiquement, les paramètres mécaniques recueillis, notamment les valeurs de déformations pour lesquelles l'os commence à se fissurer, sont intégrés dans les simulations numériques. Un scénario simplifié du remodelage osseux est alors mis en place au sein des microstructures étudiées expérimentalement et, par ailleurs, supposées endommageables. Une loi d'évolution de l'endommagement est introduite et fait l'objet d'un travail d'homogénéisation temporelle afin de considérer l'endommagement par fatigue. Les facteurs d'activation du remodelage et l'évolution des champs mécaniques au cours du processus sont, en particulier, étudiés. L'interaction du phénomène biologique et du comportement mécanique, à l'échelle de l'ostéon, est ainsi mise en évidence. / The understanding of the cortical bone remodelling process at the microscopic scale is essential in the prediction of the risk of fracture. Indeed, bone remodelling allows the perpetual regeneration of damage or old bone. The determining factors as well as the consequences of the phenomenon on the mechanical parameters of the microstructure are assessed. An experimental and numerical approach is proposed. Eight femurs from old women are analysed. Experiments are achieved at different scales. At the macroscopical scale, the Young modulus and the fracture parameters are estimated through compression testing and their eventual relations with the morphometrical characteristics (age, porosity and mineral density) are checked. Analyses of the local deformation evolution and of nanoindentation tests give access to the micro- and nanoscales and reveal the bone heterogeneity. Bone damage, especially the stage of microcracks initiation and the heterogeneity of the Young modulus as well as the mineral density are assessed. Macroscopically, porosity is determining. Microscopically, the mechanical values ob- tained, particularly the deformation value at the stage of microcracks initiation, are implemented in the numerical simulation. A bone remodelling scenario is carried out in the former experimental microstructures, supposed damageable. A damage evolution law is set and is improved by taking into account the fatigue damage through a time homogenization method. The factors of remodelling activation and the mechanical parameters evolution during the remodelling process are investigated. Eventually, the interaction between the biological phenomenon and the mechanical behaviour, at the osteon scale, is revealed.
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In vitro effects of palmitoleic acid on osteoblast differentiation in MG-63 osteosarcoma cells and human adipose-derived stromal cellsHoward, Kayla January 2019 (has links)
Bone is an important organ influenced by mechanical load, hormones, nutrition and disease. During bone remodelling, osteoclasts resorb bone and osteoblasts form new bone. Osteoblasts are derived from mesenchymal stem cells (MSCs) such as adipose-derived stromal cells (ASCs). The mitogen-activated protein kinase (MAPK) pathway has been shown to interfere with osteoblast differentiation from an early stage. Runt related transcription factor 2 (RUNX2) exerts an effect downstream from p38 MAPK. RUNX2 phosphorylation by p38 MAPK may increase osteoblast differentiation markers such as alkaline phosphatase (ALP), osteoprotegerin (OPG) and receptor activator of nuclear factor kB ligand (RANKL). Palmitoleic acid (PLA), an omega-7 monounsaturated fatty acid (MUFA), promotes anti-osteoclastogenic effects, however, the effects of PLA on osteoblasts has not been reported.
Osteoporosis is a condition which has debilitating effects in the elderly. Unsaturated fatty acids (UFA) have been studied for their beneficial effects on human health for a number of years. Polyunsaturated fatty acids (PUFA) have been studied as a potential therapeutic agent to prevent and assist in managing the condition. Few studies have been conducted on the effects of MUFA on bone therefore this study aimed to investigate the effects of PLA on osteoblast differentiation using ASCs and MG-63 osteosarcoma cells as an osteoblast model.
ASCs and MG-63 osteosarcoma cell lines were exposed to PLA (20-100 μM) in osteogenic media (OM). The effects of PLA on cell viability was evaluated on undifferentiated cells. Thereafter, cells were exposed to PLA for 7, 14 or 21 days. Subsequently ALP activity, calcium mineralisation, gene expression, protein expression and adipogenesis were assessed.
In this study, PLA had no significant effects on cell viability in undifferentiated cells. Furthermore, PLA had no significant effects on ALP activity, calcium mineralisation or phosphorylated extracellular signal-regulated kinase (pERK)/extracellular signal-regulated kinase (ERK) expression in differentiating cells, however, ALP activity increased at 7 day in ASCs and 21 days in MG-63 cells. Alizarin Red S staining increased at 21 days in both cell lines with a significant increase in the ASCs, however, calcium nodules were not visible. In the ASCs, PLA significantly increased the gene expression of ALP at 7 and 14 days compared to control (p<0.01 and p<0.05) while RANKL was significantly decreased at 7 days compared to the control (p<0.05). In the MG-63 cells, RUNX2 and OCN were significantly reduced at 7 days compared to control (p<0.05) and ALP, RUNX2, Osx and RANKL were significantly reduced at 14 days compared to control (p<0.001 and p<0.05). In the ASCs, lipid accumulation was not present after 21 days while in MG-63 cells, there was a significant increase in lipid accumulation at a high concentration of PLA after 21 days compared to control (p<0.05).
This is the first study to explore the effects of PLA on osteoblast formation using ASCs and MG-63 osteosarcoma cells. Results suggest that PLA exerted changes in the ASCs and MG-63 cells during osteoblast differentiation, however, these changes were not significant. To conclude, PLA showed some significant effects on osteoblast-specific gene expression, however, most of the osteoblast-specific gene expression was downregulated, particularly in the MG-63 cells, after PLA treatment. / Dissertation (MSc)--University of Pretoria, 2020. / Physiology / MSc / Unrestricted
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Avaliação histomorfométrica, imunoistoquímica e microtomográfica da ação da terapia laser de baixa potência no processo de reabsorção radicular durante movimentação ortodôntica induzida em ratos / Histomorphometric, immunohistochemistry and microtomography evaluation of the effect of low level laser therapy in root reabsorption process during orthodontic movement induced in ratsSuzuki, Selly Sayuri 01 June 2016 (has links)
A movimentação dentária induzida é um processo biológico complexo mediado por estímulos mecânicos, levando a um subsequente processo de remodelação óssea, podendo haver reabsorção indesejada da raiz dentária provocada pelo excesso de força. Uma vez que a movimentação ortodôntica se baseia em um processo inflamatório localizado, o propósito deste estudo foi avaliar os efeitos do laser de baixa potência no processo de remodelação óssea e reabsorção radicular, buscando correlacionar as mudanças metabólicas observadas a nível celular ocorridas nos dias iniciais da movimentação dentária às alterações teciduais observadas microscopicamente e à arquitetura e morfologia do trabeculado e cortical ósseo. Primeiros molares de sessenta e oito ratos machos Wistar foram submetidos à movimentação induzida, divididos em 3 grupos: controle negativo (nenhuma movimentação), não irradiado (movimentação sem irradiação) e laser (movimentação e irradiação com laser de baixa potência de comprimento de onda de 810 nm, potência de 100 mW, área de 0,02cm2 e energia de 1,5 J/ponto) e eutanasiados nos dias 3, 6, 9, 14 e 21. Mensurações da movimentação dentária e análises histomorfométricas foram realizadas em todos os dias estudados. Análise imunoistoquímica dos marcadores RANKL, OPG e TRAP e avaliações por microscopia eletrônica de varredura (MEV) foram feitas nos dias 3, 6 e 9 e o ensaio Western Blotting para proteínas RANKL e SOFAT e imagens de microtomografia computadorizada (MicroCT) nos dias 14 e 21. Os resultados deste estudo mostraram que a movimentação dentária foi significantemente maior no grupo Laser (aumento em média de 40%) em todos os dias avaliados. O lado de compressão mostrou maior expressão de RANKL e osteoclastos TRAP-positivos nos dias 3, 6 e 9 (p<0,05), promovendo significativa redução na área de osso alveolar presente no lado de compressão nos dias 6, 9 e 14 (p<0,05), e alterações microestruturais, como menor fração de volume ósseo/volume total, menor densidade óssea mineral aos 14 dias. A irradiação com laser também aumentou a expressão de RANKL e a citocina SOFAT no dia 14. No lado de tensão, houve maior expressão de OPG especialmente aos 9 dias (p<0,001) e significativo aumento na área de osso alveolar presente nos dias 14 (p<0,01) e 21 (p<0,05) histomorfometricamente e maior densidade óssea mineral e espessura das trabéculas aos 21 dias (p<0,01). Com relação às áreas de hialinização presentes, os resultados mostraram áreas significantemente reduzidas nos dias 3, 6 e 9 nos grupos irradiados, o que explica o menor número de odontoclastos na superfície radicular nestes dias e a redução significante das áreas de reabsorção radicular observadas nas lâminas histológicas nos dias 9, 14 e 21 e nas imagens de MEV nos dias 3 e 9. Os grupos irradiados também mostraram menor volume das lacunas de reabsorção radicular medidas no MicroCT nos dias 14 e 21, especialmente nos lados de compressão. O estudo concluiu que o laser de baixa potência influenciou a remodelação óssea, aumentou a atividade dos osteoclastos no lado da compressão, e estimulou a formação óssea no de tensão, acelerando significativamente o movimento dentário e potencialmente reduzindo as áreas de necrose no ligamento periodontal e, consequentemente, a reabsorção radicular. / Tooth movement is a complex biological process induced by mechanical stimulation, leading to a subsequent process of bone remodeling, concomitantly unwanted root resorption may occur caused by excessive force. Since orthodontic movement is based on a localized inflammatory process, the purpose of this study was to evaluate the effects of low-level laser therapy on the process of bone remodeling and root resorption, searching to correlate metabolic changes observed at cellular level in the initial days of tooth movement to tissue changes observed microscopically and both architecture and morphology of trabecular and cortical bone. Upper first molars of sixtyeight male Wistar rats were submitted to induced movement, divided into 3 groups: negative control (no movement), non-irradiated (movement without irradiation) and Laser (movement and irradiation using low level laser of 810 nm wavelength, 100 mW power, 0.02cm2 area, energy of 1.5J/point) and euthanized on days 3, 6, 9, 14 and 21. Measurements of tooth movement and histomorphometric analysis were performed at all days. Immunohistochemistry analysis of RANKL, OPG and TRAP markers and scanning electron microscopy (SEM) were made on days 3, 6 and 9. Western Blotting method to evaluate RANKL and SOFAT proteins and MicroCT images were performed on days 14 and 21. The results of this study showed that tooth movement was significantly greater in the irradiated side (increased in average of 40%) in all evaluated days. The compression side showed higher expression of RANKL and TRAP-positive osteoclasts on days 3, 6 and 9 (p <0.05), promoting significant reduction in alveolar bone area in the compression side on days 6, 9 and 14 ( p <0.05), and leading to microstructural changes such as decrease of the fraction of bone volume / total volume (BV/TV) and the bone mineral density (BMD) at 14 days. The laser also increased RANKL expression and SOFAT on day 14. On the tension side there was an increased expression of OPG especially after 9 days (p <0.001), a significant increase in alveolar bone area on days 14 (p < 0.01) and 21 (p <0.05) histomorphometrically and increase in bone mineral density and trabecular thickness after 21 days (p <0.01). Regarding hyalinized areas, the results showed significant reduced areas on days 3, 6 and 9 in irradiated groups, which explains the lower number of clastic cells on the root surface in these days, and a significant reduction of areas of root resorption observed in histology on days 9, 14 and 21 and on days 3 and 9 by SEM images. Irradiated groups also showed less volume of root resorption lacunaes measured by MicroCT on days 14 and 21, especially in the compression side. The study concludes that the low-level laser therapy had an effect on bone remodeling, increasing osteoclast activity on the compression side, and stimulating bone formation in tension side, resulting in significant tooth movement acceleration and potentially reducing the areas of necrosis in the periodontal ligament and consequently the root resorption process.
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Growth modification of the temporomandibular joint by functional appliances: a histomorphometric study using sheepMa, Bingkui January 2002 (has links)
In order to investigate growth modifications of the temporomandibular joint (TMJ) during dentofacial orthopaedic treatment, various functional appliances have been used to prompt the mandible into a protrusive position in various animal experimental models. The general purpose of this project was (i) to test the effectiveness of a functional appliance specially designed for sheep; (ii) to clarify whether or not forward mandibular displacement in sheep is associated with faster and/or redirected condylar growth; (iii) to evaluate the sheep as a model for dentofacial orthopaedic research by comparing the similarities of mandibular condylar growth in sheep and humans; (iv) to detail the position of the mandible during forward mandibular posturing and the effects of mandibular forward displacement on modelling and remodelling of the mandibular condyle. The specific purpose of this project was to reveal whether functional appliance treatment increases the quantity of bone formed during the treatment, or changes the distribution of the bone, or both. Eight, 4-month old, castrated male Merino sheep were randomly assigned to experimental or control groups with 4 in each group. Cast functional appliances were fabricated for the animals in the experimental group. The treatment period was 15 weeks. Calcein (day 1) tetracycline (13 weeks) and alizarin red S (3 days before sacrifice) fluorochromes were administered to all animals. Dental casts, endosseous implant markers and cephalograms were used to analyse the 3-D displacement of the mandible. Undecalcified mid-sagittal sections of TMJ were used to evaluate the tissue responses induced by the appliances. Dynamic parameters of bone formation, static indices of bone-forming and resorbing activity as well as structural indices of trabecular bone were estimated using histomorphometry. The trabecular bone was sampled from two regions: (i) a subchondral region; (determined by 2nd and 3rd labels), believed to comprise bone newly-formed during the experimental period; and (ii) a central region (labelled by all the three fluorochromes), believed to comprise bone which existed before the experiment. The cortical bone was divided into anterior and posterior regions for analysis. The weight of the animals was measured monthly to monitor their growth. Metacarpus growth was also evaluated. During the experimental period, the animals were found to maintain their weight within the normal range and grew normally. The appliance was found to displace the mandible to a downward and forward position with a net condylar displacement of 2.4 mm. The observed adaptive responses in the TMJ induced by the appliances included; the condylar process was less tapered and rounder in the experimental group than in the controls, and anteriorly thickened condylar cartilage and a thickened compact bone layer along the anterior surface of the posterior wall of the glenoid fossa. The mandibular condylar growth vector in sheep was found to be in a postero-superior direction. Condylar growth in the control sheep during the experimental period varied from 8.8 to 11.9 mm, with the mean being 10.6 mm, which is quantitatively similar to two years of condylar growth in human adolescents. In the experimental sheep, the condylar growth varied from 8.5 to 13.3 mm, with the mean being 11.4 mm. When metacarpal growth and weight gain were taken into consideration using multivariant analysis, the coefficients for growth in the postero-superior and posterior direction were found to be high, with adjusted r2 as 0.84 and 0.82 respectively. The induced condylar growth was estimated to be largest in the posterior direction (2.3 mm), which is also similar to previous reports in humans. Regional differences in adaptive response within the mandibular condyle were found in this study. In the experimental group, bone volume fraction (BV/TV) of the subchondral regions decreased, although the specific bone surface and bone formation rates increased. This low BV/TV was associated with decreased trabecular thickness and increased trabecular separation. In the central region of the experimental group's condyle, BV/TV was unchanged. However, an increased osteoid surface (OS/BS) was defined when the eroded surface (ES/BS) was taken into consideration. The sheep were found to cope well with the experimental procedures and the appliance used in this study has been effective in inducing adaptive responses in the TMJ. Consequently, it is believed that the sheep is an appropriate animal model for quantitative histological analysis of the responses to functional appliance treatment. The first null hypothesis, that functional appliance treatment has no effect on bone matrix mineralisation was rejected. The second null hypothesis, functional appliance treatment has no effect on the mineralisation lag time, was rejected. The results indicated that the treatment effects of functional appliances involve reorganisation of the TMJ through bone modelling and remodelling. An important mechanism of functional appliance treatment is, therefore, suggested to be a change in the distribution of bone rather than an increase in the quantity of bone. Posterior rotation of the principle tensile strain angle (Et) suggested an posteriorly altered direction of the condylar growth. Increased new bone formation in the glenoid fossa suggested an anterior re-positioning of the temporomandibular joint. / Thesis (Ph.D.)--Dental School, 2002.
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Osteoporosis: An Age-Related and Gender-Specific Disease – A Mini-ReviewPietschmann, Peter, Rauner, Martina, Sipos, Wolfgang, Kerschan-Schindl, Katharina 24 February 2014 (has links) (PDF)
Osteoporosis, a classical age-related disease and known to be more common in women than in men, has been reported increasingly often in men during the past few years. Although men at all ages after puberty have larger bones than women, resulting in greater bending strength, mortality after a hip fracture, one of the major complications of osteoporosis, is more common in men than in women. Sex hormone deficiency is associated with unrestrained osteoclast activity and bone loss. Even though estrogen deficiency is more pronounced in women, it appears to be a major factor in the pathogenesis of osteoporosis in both genders. In contrast to osteoporosis in postmenopausal women, the treatment of osteoporosis in men has been scarcely reported. Nevertheless, some drugs commonly used for the treatment of osteoporosis in women also appear to be effective in men. The aim of this study is to review primary osteoporosis in the elderly with particular emphasis on gender-related aspects. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.
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Phytoestrogen status in relation to sociodemographic factors and biomarkers of bone health in older Brisbane womenHanna, Katherine Lavina January 2006 (has links)
Background: Phytoestrogens are diphenolic compounds found in plants with a structure and molecular weight similar to oestradiol which enables them to bind to the oestrogen receptor. Isoflavonoids occur mainly within the legume family with highest concentration in soybeans. Lignans are found in a range of plant foods and the richest known source is linseed. Few studies have been published on intake of isoflavonoids and none were located on intake of lignans in Australian women. The validity of methods designed to estimate intake can be assessed using urinary excretion of isoflavonoids and lignans as studies have found an association between intake and excretion of isoflavonoids and lignans. It has been proposed that, through their ability to act like oestrogen, phytoestrogens could decrease bone turnover and attenuate the loss of bone mineral density (BMD) at menopause. The aims of this research were to determine the pattern of intake of isoflavonoids and lignans in 500 women from food and supplements and to assess a questionnaire used to estimate intake using excretion in a sub-sample of 141 women. Associations between usual intake or excretion of isoflavonoids and lignans and biomarkers of bone health were also examined. Methods: A cross-sectional study was conducted involving 500 women aged 40-80 years participating in the Longitudinal Assessment of Ageing in Women (LAW), a 5 year study being conducted in the Betty Byrne Henderson Centre at the Royal Brisbane and Women's Hospital. Subjects were randomly selected from the electoral role and stratified into ten year age groups. Intake of isoflavonoids and lignans from food and supplements was assessed using a specially designed questionnaire containing 110 items. Values for individual items were obtained from published literature and summed to provide average daily intakes of isoflavonoids and lignans (mg/d). A sub-sample of 141 women was recruited to take part in the assessment of the association between phytoestrogen intake and excretion. Participants collected three 24-h urine samples spaced over one week. Samples were analysed using high performance liquid chromatography MS/MS for seven isoflavonoids and four lignans. Bone mineral densities (BMD) of the femur neck, total hip and lumbar spine were measured by dual energy x-ray absorptiometry. Bone formation was assessed using serum bone alkaline phosphatase (bone ALP) and osteocalcin (OC) and bone resorption was assessed using deoxypyridinoline (DPD) and urinary excretion of N-terminal cross-linking telopeptide of type-I collagen (NTX). Potential confounding factors were also evaluated. Statistical analyses were conducted using SPSS for windows (version 10). Participants were defined as consumers if they reported intake of one or more serves of soy or linseed in the prior month. Differences in socio-demographic and lifestyle characteristics between groups were assessed using ANOVA and Chi Square tests. Associations between intake and excretion of phytoestrogens were assessed using Spearman's rank-order correlations () for non-normal data. Phytoestrogen intake was categorised into four groups for the assessment of the association with markers of bone health. Associations between phytoestrogen excretion and markers of bone health were assessed using Pearson's product moment correlations for normal data (r) and Spearman's rank-order correlations for non-normal data. A value of P < 0.05 was taken as statistically significant. Results: Consumption of soy food was reported by 40% and consumption of linseed by 34% of women. Median (range) intakes among soy/linseed consumers for isoflavonoids, 3.87 (0-173) mg/d, and lignans, 2.40 (0.1-33) mg/d, were significantly higher than corresponding intakes among non consumers of 0.005 (0-2.6) and 1.57 (0.4-4.7) mg/d, respectively (P < 0.001). Soy/linseed consumers reported higher intakes of energy (P=0.043), dietary fibre (P=0.003) and polyunsaturated fat (P=0.004); and a higher level of physical activity (P=0.006), SEP (P < 0.001), education (P < 0.001) and supplement use (P < 0.001). Use of non-prescription supplements for menopause in the previous month was reported by 13% of women. A review of supplements available for treatment of menopause indicated that use of soy, red clover, black cohosh and sage could have a role in treatment of menopause symptoms. Evidence supporting the presence of oestrogenic components was available for soy and red clover isoflavonoids only. There was a significant association between intake and excretion of isoflavonoids within the total group (r=0.207, P < 0.05), with a stronger association in soy consumers (r=0.364, P < 0.01). Excretion of isoflavonoids was detected in women who did not report known intake of soy foods, suggesting isoflavonoids could be derived in small amounts from other plant foods or use of soy as an ingredient in processed foods. There was no significant association between intake and excretion of lignans, however both intake and excretion were associated with dietary fibre (r=0.303 and r=0.230, respectively, P < 0.01 for both). Bone ALP was higher among the very low isoflavonoid intake group (P=0.005) for the total sample (P=0.005) and women with BMI≤25 kg/m2 (P=0.002). Data also demonstrated an inverse association between excretion of isoflavonoids and NTX within women with BMI≤25 kg/m2 (r=-0.33, P < 0.05). There was a positive association between lignan excretion and bone ALP in the total sample (r=0.21, P < 0.05) which was strengthened in women with osteoporosis/osteopenia (r=0.41, P < 0.05) and a positive association between lignan excretion and DPD among women with BMI≤25 kg/m2 (ρ=0.28, P < 0.05) All associations remained significant after adjustment for confounding. Conclusions: Few women who chose phytoestrogen-rich foods consumed amounts similar to women with traditional soy-based diets although some achieved high intakes with supplements. Women who consumed soy or linseed foods differed in lifestyle and sociodemographic characteristics that could influence the association with disease in epidemiological studies. Results indicated that the phytoestrogen questionnaire was useful for assessment of isoflavonoids but was not acceptably precise for measurement of lignans. Findings suggest that there is an inverse association between isoflavonoid status and bone ALP and NTX although the precise mechanism of action has not been clarified. The association between lignan intake and bone is less well understood; however findings of a positive association with bone ALP indicate that further research on the lignan content of foods and the inclusion of lignans in studies is warranted.
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Desenvolvimento de uma metodologia para analise de bioengenharia em ossos compactos com remodelagem superficial pelo metodo dos elementos de contorno 3D em meios transversalmente isotropicos / Development of a methodology for bioengineering analysis of compact bones with surface remodeling using 3D boundary element method in transversely isotropic mediaNoritomi, Pedro Yoshito 07 August 2005 (has links)
Orientador: Paulo Sollero / Tese (doutorado) - Universidade Estadual de Campinas. Faculdade de Engenharia Mecanica / Made available in DSpace on 2018-08-06T09:16:06Z (GMT). No. of bitstreams: 1
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Previous issue date: 2005 / Resumo: Este trabalho mostra o desenvolvimento de uma metodologia para análise de problemas de bioengenharia, aplicando modelagem numérica elastostática de tensões e deformações, baseada no método dos elementos de contorno com formulação 3D para meios transversalmente isotrópicos lineares, incluindo a capacidade de simulação do comportamento de remodelagem óssea superficial. A implementação do núcleo transversalmente isotrópico sobre a estrutura básica de análise por elementos de contorno 3D utilizou a solução fundamental proposta por Pan & Chou e revisada por Loloi, tendo exigido o cálculo adicional das soluções fundamentais de força de superfície a partir da derivação das soluções fundamentais de deslocamento. O modelo de remodelagem óssea superficial baseou-se na hipótese de estímulo biológico por campo de deformação, partindo de um modelo 2D, adaptado para o espaço 3D com o uso de deformações principais como grandezas de referência. As implementações foram testadas através de análises numéricas de problemas com solução analítica e validações com resultados de aplicações comerciais baseadas em elementos finitos, para problemas padrão de engenharia, bem como comparações com resultados da literatura para problemas de bioengenharia. A análise dos resultados mostrará que, tanto a metodologia quanto as implementações são funcionais, oferecendo uma base sólida para desenvolvimento e teste de novas soluções de bioengenharia / Abstract: This work shows the development of a methodology to analyse bioengineering problems using elastostatic stress-strain numerical modeling based on a 3D transversely isotropic linear boundary element formulation including surface bone remodeling simulation capabilities. The transversely isotropic kernel implementation on the basic 3D boundary element analysis program used the fundamental solution purposed by Pan & Chou and revised by Loloi, with additional fundamental solutions for traction calculation made with the displacement fundamental solution derivatives. The surface bone remodeling model was based on a 2D strain field biological stimulus, extended to the 3D space by using the principal strain as reference values. The implementations were tested through numerical analysis of problems with analytical solution and validation with commercial finite elements applications for standard engineering problems, as well as comparison with literature data for bioengineering problems. The analysis of results will show that both, the methodology and the implementations are fully functional, offering a solid start for development and test of new bioengineering solutions / Doutorado / Mecanica dos Sólidos e Projeto Mecanico / Doutor em Engenharia Mecânica
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Avaliação histomorfométrica, imunoistoquímica e microtomográfica da ação da terapia laser de baixa potência no processo de reabsorção radicular durante movimentação ortodôntica induzida em ratos / Histomorphometric, immunohistochemistry and microtomography evaluation of the effect of low level laser therapy in root reabsorption process during orthodontic movement induced in ratsSelly Sayuri Suzuki 01 June 2016 (has links)
A movimentação dentária induzida é um processo biológico complexo mediado por estímulos mecânicos, levando a um subsequente processo de remodelação óssea, podendo haver reabsorção indesejada da raiz dentária provocada pelo excesso de força. Uma vez que a movimentação ortodôntica se baseia em um processo inflamatório localizado, o propósito deste estudo foi avaliar os efeitos do laser de baixa potência no processo de remodelação óssea e reabsorção radicular, buscando correlacionar as mudanças metabólicas observadas a nível celular ocorridas nos dias iniciais da movimentação dentária às alterações teciduais observadas microscopicamente e à arquitetura e morfologia do trabeculado e cortical ósseo. Primeiros molares de sessenta e oito ratos machos Wistar foram submetidos à movimentação induzida, divididos em 3 grupos: controle negativo (nenhuma movimentação), não irradiado (movimentação sem irradiação) e laser (movimentação e irradiação com laser de baixa potência de comprimento de onda de 810 nm, potência de 100 mW, área de 0,02cm2 e energia de 1,5 J/ponto) e eutanasiados nos dias 3, 6, 9, 14 e 21. Mensurações da movimentação dentária e análises histomorfométricas foram realizadas em todos os dias estudados. Análise imunoistoquímica dos marcadores RANKL, OPG e TRAP e avaliações por microscopia eletrônica de varredura (MEV) foram feitas nos dias 3, 6 e 9 e o ensaio Western Blotting para proteínas RANKL e SOFAT e imagens de microtomografia computadorizada (MicroCT) nos dias 14 e 21. Os resultados deste estudo mostraram que a movimentação dentária foi significantemente maior no grupo Laser (aumento em média de 40%) em todos os dias avaliados. O lado de compressão mostrou maior expressão de RANKL e osteoclastos TRAP-positivos nos dias 3, 6 e 9 (p<0,05), promovendo significativa redução na área de osso alveolar presente no lado de compressão nos dias 6, 9 e 14 (p<0,05), e alterações microestruturais, como menor fração de volume ósseo/volume total, menor densidade óssea mineral aos 14 dias. A irradiação com laser também aumentou a expressão de RANKL e a citocina SOFAT no dia 14. No lado de tensão, houve maior expressão de OPG especialmente aos 9 dias (p<0,001) e significativo aumento na área de osso alveolar presente nos dias 14 (p<0,01) e 21 (p<0,05) histomorfometricamente e maior densidade óssea mineral e espessura das trabéculas aos 21 dias (p<0,01). Com relação às áreas de hialinização presentes, os resultados mostraram áreas significantemente reduzidas nos dias 3, 6 e 9 nos grupos irradiados, o que explica o menor número de odontoclastos na superfície radicular nestes dias e a redução significante das áreas de reabsorção radicular observadas nas lâminas histológicas nos dias 9, 14 e 21 e nas imagens de MEV nos dias 3 e 9. Os grupos irradiados também mostraram menor volume das lacunas de reabsorção radicular medidas no MicroCT nos dias 14 e 21, especialmente nos lados de compressão. O estudo concluiu que o laser de baixa potência influenciou a remodelação óssea, aumentou a atividade dos osteoclastos no lado da compressão, e estimulou a formação óssea no de tensão, acelerando significativamente o movimento dentário e potencialmente reduzindo as áreas de necrose no ligamento periodontal e, consequentemente, a reabsorção radicular. / Tooth movement is a complex biological process induced by mechanical stimulation, leading to a subsequent process of bone remodeling, concomitantly unwanted root resorption may occur caused by excessive force. Since orthodontic movement is based on a localized inflammatory process, the purpose of this study was to evaluate the effects of low-level laser therapy on the process of bone remodeling and root resorption, searching to correlate metabolic changes observed at cellular level in the initial days of tooth movement to tissue changes observed microscopically and both architecture and morphology of trabecular and cortical bone. Upper first molars of sixtyeight male Wistar rats were submitted to induced movement, divided into 3 groups: negative control (no movement), non-irradiated (movement without irradiation) and Laser (movement and irradiation using low level laser of 810 nm wavelength, 100 mW power, 0.02cm2 area, energy of 1.5J/point) and euthanized on days 3, 6, 9, 14 and 21. Measurements of tooth movement and histomorphometric analysis were performed at all days. Immunohistochemistry analysis of RANKL, OPG and TRAP markers and scanning electron microscopy (SEM) were made on days 3, 6 and 9. Western Blotting method to evaluate RANKL and SOFAT proteins and MicroCT images were performed on days 14 and 21. The results of this study showed that tooth movement was significantly greater in the irradiated side (increased in average of 40%) in all evaluated days. The compression side showed higher expression of RANKL and TRAP-positive osteoclasts on days 3, 6 and 9 (p <0.05), promoting significant reduction in alveolar bone area in the compression side on days 6, 9 and 14 ( p <0.05), and leading to microstructural changes such as decrease of the fraction of bone volume / total volume (BV/TV) and the bone mineral density (BMD) at 14 days. The laser also increased RANKL expression and SOFAT on day 14. On the tension side there was an increased expression of OPG especially after 9 days (p <0.001), a significant increase in alveolar bone area on days 14 (p < 0.01) and 21 (p <0.05) histomorphometrically and increase in bone mineral density and trabecular thickness after 21 days (p <0.01). Regarding hyalinized areas, the results showed significant reduced areas on days 3, 6 and 9 in irradiated groups, which explains the lower number of clastic cells on the root surface in these days, and a significant reduction of areas of root resorption observed in histology on days 9, 14 and 21 and on days 3 and 9 by SEM images. Irradiated groups also showed less volume of root resorption lacunaes measured by MicroCT on days 14 and 21, especially in the compression side. The study concludes that the low-level laser therapy had an effect on bone remodeling, increasing osteoclast activity on the compression side, and stimulating bone formation in tension side, resulting in significant tooth movement acceleration and potentially reducing the areas of necrosis in the periodontal ligament and consequently the root resorption process.
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Die Wirkung von systemisch appliziertem rekombinantem speziesspezifischem Wachstumshormon auf den Knochenumbau am AchsenskelettFunk, Julia Franziska 27 October 2004 (has links)
Die Auswirkung der systemischen Applikation von 100µg/kg/KG rekombinantem speziesspezifischem Wachstumshormon (GH) über 26 Tage auf den Knochenumbau des Yucatan-Minischweins wurde an Wirbelkörpern, Beckenkämmen und Rippen histomorphometrisch untersucht. Der Einfluss der Distraktionsosteogenese auf die Mineralappositionsrate wurde ermittelt. 24 Tiere wurden an der linken Tibia osteotomiert, ein Fixateur externe wurde zur Distraktionsosteogenese von 2mm/d angelegt. 12 Tiere erhielten Wachstumshormon, 12 Placebo. Eine polychrome Fluoreszenzmarkierung wurde i.v. appliziert. Post mortem wurden Schnitte und Dünnschliffe der genannten Knochen angefertigt. Mittels Bildanalysesystem erfolgte die histomorphometrische Auswertung anhand der Parameter Knochendichte, Porosität, Trabekeldicke, Trabekelumbauparameter und Mineralappositionsrate (MAR). Signifikante Veränderungen durch Wachstumshormongabe konnten gezeigt werden. Unterschiede zwischen den Lokalisationen traten auf. So war die Knochendichte der GH-Tiere an Wirbelkörpern und Rippen signifikant geringer als in der Placebogruppe, kein signifikanter Unterschied zeigte sich am Beckenkamm. An allen 3 Lokalisationen war in der Wachstumshormongruppe eine signifikant erhöhte kortikale Porosität zu finden. Nur am Wirbelkörper waren die trabekuläre Knochendichte und die Trabekeldicke bei den GH-Tieren signifikant geringer als bei den Kontrolltieren. Die Knochenresorption war in der GH-Gruppe am Beckenkamm signifikant erhöht. Die Formation war bei den GH-Tieren sowohl iliakal als auch vertebral signifikant erhöht. Insgesamt ergab sich damit ein signifikant verminderter Anteil an ruhender Oberfläche bei den GH-Tieren. In ihrer Ausprägung waren diese Parameter zwischen Wirbel und Beckenkamm signifikant verschieden. Die MAR war in der GH-Gruppe signifikant höher als in der Placebogruppe, außerdem war sie während der Distraktionsphase signifikant höher als während der Konsolidierungsphase. Vergleicht man die Parameter mit Angaben aus der Literatur, kann bestätigt werden, dass die hier verwendeten Messmethoden geeignet sind, und dass das Yucatan-Minischwein ein geeignetes Modell zur Klärung von Fragestellungen der Knochenbiologie auch im Hinblick auf den Vergleich zum Menschen darstellt. / The effect of systemic application of 100 µg/kg recombinant species specific growth hormone (GH) on the bone remodelling of 24 Yucatan mini pigs for a duration of 26 days was evaluated on vertebral body, iliac crest, and rib. The influence of distraction osteogenesis on the mineral apposition rate (MAR) was examined. 24 animals were osteotomized at the left tibia and treated with an external fixator for distraction osteogenesis of 2 mm/d. 12 of them received GH, the others placebo. A sequential polychrome fluorescent labelling was applied. After sacrifice thin and ground sections were manufactured. Using an image analysis system, a histomorphometrical evaluation was performed by means of bone density, porosity, trabecular thickness, trabecular remodelling parameters, and mineral apposition rate (MAR). Significant changes after GH application were visible. Differences between the locations were found. The bone density of vertebrae and ribs of the treatment group was significantly lower than that of the placebo group, whereas no difference was seen in the iliac crest biopsies. All 3 locations showed a significantly higher cortical porosity in the GH group. The trabecular bone density and the trabecular thickness were significantly decreased only in the vertebral bodies of the treatment group. The bone resorption was significantly elevated in the iliac crests of the GH group. The bone formation was increased in the iliac crest as well as in the vertebrae of the GH treated animals. Therefore, the resting surface was significantly decreased in the GH group. The extent of these effects was significantly different between iliac crest and vertebral body. The MAR was significantly higher in the GH group and during distraction osteogenesis. Compared to results of other study groups these parameters validate the methods of measurement and show that the Yucatan mini pig is a suitable model for research concerning bone biology, especially concerning the comparison with human parameters.
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