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Avaliação da reparação óssea em defeitos críticos após a aplicação da proteína rhBMP-2 pura e/ou combinada em animais normais e sob efeito do alcoolismo crônico / Assessment of the bone healing process in critical defects after application of the rhBMP-2 pure protein and / or combined in normal animals and under the effect of chronic alcoholismKotake, Bruna Gabriela dos Santos 19 October 2012 (has links)
O alcoolismo é considerado um redutor da formação óssea, podendo levar a distúrbios osteometabólicos, já a rhBMP-2 é uma proteína morfogenética conhecida por desempenhar um papel importante nos processos de reparação e indução da formação óssea. O objetivo deste estudo foi investigar a ação do alcoolismo e a resposta do reparo em defeitos ósseos (DO) na calvária de ratos, após a aplicação da rhBMP-2, pura e combinada a uma matriz de colágeno. Foram utilizados 80 ratos divididos em 8 grupos, cada um deles com um período de espera até o sacrifício de 4 e 6 semanas, após a criação cirúrgica do defeito ósseo na calvária de 5 mm. Os grupos foram divididos em G1) água \"ad libitum\" + DO, G2) álcool \"ad libitum\" + DO, G3) água + DO + 5μg rhBMP-2 pura, G4) álcool + DO + 5μg rhBMP-2 pura, G5) água + DO + carreador esponja de colágeno, G6) álcool + DO + carreador esponja de colágeno, G7) água e + DO + 5μg rhBMP- 2/esponja de colágeno, G8) álcool + DO + 5μg rhBMP-2/esponja de colágeno. Os dados radiográficos e os dados para a análise de fibras colágena tipo I e III foram submetidos à análise estatística Kruskal-Wallis e Dunn\'s Multiple, os dados histológicos ao teste de análise de variância (ANOVA) e Tukey. O resultado encontrado para a análise radiográfica no tempo de 6 semanas demonstrou que os grupos tratados com rhBMP-2 independente da utilização do carreador e do etanol apresentaram maior neoformação óssea (p<0,05), para o tempo de 4 semanas não foi encontrada diferença estatística. Para a análise imunoistoquímica, a ostecalcina (OC) e sialoproteína óssea (BPS) foram predominantes nos grupos tratados com rhBMP-2. Para a análise histológica, a quantificação de fibras colágenas tipo I apresentou diferença estatística entre os grupos (p<0,05), com aumento destas fibras principalmente nos grupos tradados com rhBMP-2 associado a esponja de colágeno; e na análise quantitativa por estereologia, o volume de tecido ósseo neoformado foi maior para os grupos tratados com rhBMP-2 pura ou associada ao carreador, porém para os grupos tratados com etanol, houve maior neoformação óssea para o grupo tratado com rhBMP-2 associado ao carreador nos períodos de 4 e 6 semanas (p<0,001). Concluiu-se neste modelo experimental que, o carreador foi efetivo na bioliberação da rhBMP-2, mesmo na presença do etanol. / Alcoholism is considered a reducer for new bone formation, may leading to osteometabolic disturbance, considering that the rhBMP-2 is a morphogenetic protein known to play an important role in the bone healing processes. The aim of this study was to investigate the action of alcoholism and its effect on the repair of bone defects (DO) performed in rat calvaria after the rhBMP-2 application, pure or combined with a collagen matrix. We used 80 rats divided into eight groups, each one with a waiting period for sacrifice of 4 and 6 weeks after the bone defect (5mm) delineation in the rat skull. The groups were divided into: G1) water \"ad libitum\" + DO, G2) alcohol \"ad libitum\" + DO, G3) water + DO + 5μg pure rhBMP-2, G4) alcohol + DO + 5μg pure rhBMP-2, G5 ) DO + water + collagen sponge carrier, G6) alcohol + DO + collagen sponge carrier, G7) and water + DO + 5μg rhBMP-2/collagen sponge, G8) alcohol + DO + 5μg of rhBMP-2/collagen sponge. The radiographic data were submitted to statistical analysis Kruskal-Wallis and Dunn\'s Multiple, histological data to analysis of variance (ANOVA) and Tukey test. Radiographically, it was found after 6 weeks that the groups treated with rhBMP-2, independently of the carrier use and ethanol administration, more new bone formation (p<0.05), at 4 weeks it was not found statistical difference. For immunohistochemical analysis, ostecalcin (OC), and bone sialoprotein (BPS) were predominant in groups treated with rhBMP-2. For histological quantification of collagen type I fibers, statistical difference between groups was found (p<0.05) with the increasing of these fibers in the groups treated with rhBMP-2 combined with collagen sponge; and quantitative by stereology, aiming to calculate the volume of new bone, it was found higher values for the groups treated with rhBMP-2 pure or combined to the carrier; but for the groups treated with ethanol, it was found higher bone formation in the groups treated with rhBMP-2 associated to the carrier in the periods of 4 and 6 weeks (p <0.001). It was concluded in this experimental model that the carrier was effective for rhBMP-2 delivery, even in the presence of ethanol.
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Avaliação da reparação óssea em defeitos críticos após a aplicação da proteína rhBMP-2 pura e/ou combinada em animais normais e sob efeito do alcoolismo crônico / Assessment of the bone healing process in critical defects after application of the rhBMP-2 pure protein and / or combined in normal animals and under the effect of chronic alcoholismBruna Gabriela dos Santos Kotake 19 October 2012 (has links)
O alcoolismo é considerado um redutor da formação óssea, podendo levar a distúrbios osteometabólicos, já a rhBMP-2 é uma proteína morfogenética conhecida por desempenhar um papel importante nos processos de reparação e indução da formação óssea. O objetivo deste estudo foi investigar a ação do alcoolismo e a resposta do reparo em defeitos ósseos (DO) na calvária de ratos, após a aplicação da rhBMP-2, pura e combinada a uma matriz de colágeno. Foram utilizados 80 ratos divididos em 8 grupos, cada um deles com um período de espera até o sacrifício de 4 e 6 semanas, após a criação cirúrgica do defeito ósseo na calvária de 5 mm. Os grupos foram divididos em G1) água \"ad libitum\" + DO, G2) álcool \"ad libitum\" + DO, G3) água + DO + 5μg rhBMP-2 pura, G4) álcool + DO + 5μg rhBMP-2 pura, G5) água + DO + carreador esponja de colágeno, G6) álcool + DO + carreador esponja de colágeno, G7) água e + DO + 5μg rhBMP- 2/esponja de colágeno, G8) álcool + DO + 5μg rhBMP-2/esponja de colágeno. Os dados radiográficos e os dados para a análise de fibras colágena tipo I e III foram submetidos à análise estatística Kruskal-Wallis e Dunn\'s Multiple, os dados histológicos ao teste de análise de variância (ANOVA) e Tukey. O resultado encontrado para a análise radiográfica no tempo de 6 semanas demonstrou que os grupos tratados com rhBMP-2 independente da utilização do carreador e do etanol apresentaram maior neoformação óssea (p<0,05), para o tempo de 4 semanas não foi encontrada diferença estatística. Para a análise imunoistoquímica, a ostecalcina (OC) e sialoproteína óssea (BPS) foram predominantes nos grupos tratados com rhBMP-2. Para a análise histológica, a quantificação de fibras colágenas tipo I apresentou diferença estatística entre os grupos (p<0,05), com aumento destas fibras principalmente nos grupos tradados com rhBMP-2 associado a esponja de colágeno; e na análise quantitativa por estereologia, o volume de tecido ósseo neoformado foi maior para os grupos tratados com rhBMP-2 pura ou associada ao carreador, porém para os grupos tratados com etanol, houve maior neoformação óssea para o grupo tratado com rhBMP-2 associado ao carreador nos períodos de 4 e 6 semanas (p<0,001). Concluiu-se neste modelo experimental que, o carreador foi efetivo na bioliberação da rhBMP-2, mesmo na presença do etanol. / Alcoholism is considered a reducer for new bone formation, may leading to osteometabolic disturbance, considering that the rhBMP-2 is a morphogenetic protein known to play an important role in the bone healing processes. The aim of this study was to investigate the action of alcoholism and its effect on the repair of bone defects (DO) performed in rat calvaria after the rhBMP-2 application, pure or combined with a collagen matrix. We used 80 rats divided into eight groups, each one with a waiting period for sacrifice of 4 and 6 weeks after the bone defect (5mm) delineation in the rat skull. The groups were divided into: G1) water \"ad libitum\" + DO, G2) alcohol \"ad libitum\" + DO, G3) water + DO + 5μg pure rhBMP-2, G4) alcohol + DO + 5μg pure rhBMP-2, G5 ) DO + water + collagen sponge carrier, G6) alcohol + DO + collagen sponge carrier, G7) and water + DO + 5μg rhBMP-2/collagen sponge, G8) alcohol + DO + 5μg of rhBMP-2/collagen sponge. The radiographic data were submitted to statistical analysis Kruskal-Wallis and Dunn\'s Multiple, histological data to analysis of variance (ANOVA) and Tukey test. Radiographically, it was found after 6 weeks that the groups treated with rhBMP-2, independently of the carrier use and ethanol administration, more new bone formation (p<0.05), at 4 weeks it was not found statistical difference. For immunohistochemical analysis, ostecalcin (OC), and bone sialoprotein (BPS) were predominant in groups treated with rhBMP-2. For histological quantification of collagen type I fibers, statistical difference between groups was found (p<0.05) with the increasing of these fibers in the groups treated with rhBMP-2 combined with collagen sponge; and quantitative by stereology, aiming to calculate the volume of new bone, it was found higher values for the groups treated with rhBMP-2 pure or combined to the carrier; but for the groups treated with ethanol, it was found higher bone formation in the groups treated with rhBMP-2 associated to the carrier in the periods of 4 and 6 weeks (p <0.001). It was concluded in this experimental model that the carrier was effective for rhBMP-2 delivery, even in the presence of ethanol.
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Modulation of inflammatory process and tissue regeneration in calvaria mouse modelsAl-Hashemi, Jacob Yousef 17 June 2019 (has links)
MicroRNAs (miRNAs) are short, non-coding RNAs involved in the regulation of several processes associated with inflammatory diseases and infection. Bacterial infection modulates miRNA expression to subvert innate immune response. In this study, we analyzed bacterial modulation of miRNAs in bone-marrow-derived macrophages (BMMs), in which activity was induced by infection with Porphyromonas gingivalis (Pg) through a microarray analysis. Several miRNA expressions levels were modulated 3 hours post infection (at a multiplicity of infection (MOI) of 25). A bioinformatics analysis was performed to further identify pathways related to the innate immune host-response pathways that are under the influence of the selected miRNAs. To assess the effects of the identified miRNAs on cytokines secretion (pro inflammatory TNF-α and anti-inflammatory IL-10), BMMs were transfected with selected miRNAs mimics or inhibitors. Transfection with mmu-miR-155 and mmu-miR- 2137 did not modify TNF-α secretion while their inhibitors increased it. Inhibitors of mmumiR-2137 and mmu-miR-7674 increased the secretion of the anti-inflammatory IL-10. In Pginfected BMMs, mmu-miR-155-5p significantly decreased TNF-α secretion while inhibitor of mmu-miR-2137 increased IL-10 secretion. In vivo, in a Pg-induced calvarial bone resorption mouse model, injection of mmu-miR-155-5p or anti-mmu-miR-2137 reduced the size of the lesion significantly. Furthermore, anti-mmu-miR-2137 significantly reduced inflammatorycell infiltration, osteoclast activity and bone loss. Bioinformatics analysis demonstrated that pathways related to cytokines and chemokines related pathways but also osteoclast differentiation may be involved in the observed effects. The study highlights the potential therapeutic merits of targeting mmu-miR-155-5p and mmu-miR-2137 to control inflammation induced by Pg infection.
To assess the regenerative process in the same animal model, we aimed to compare the effect of Bone Morphogenic Protien 2 (BMP2), Platelets Rich Plasma (PRP), Leukocyte-Platelets Rich Fibrin (L-PRF), and Polygucosamine (PGIcNAc) on bone formation in critical size bone defects in mice. One-hundred-thirty-eight mice were divided into 23 groups (n=6), negative control, different combinations of the PGIcNAc with or without of BMP2, Collagen Sponge (SurgiFoam), PRP, and L-PRF. The 5mm defect, then, was allowed to heal. After six weeks, samples were analyzed for bone formation utilizing radiographs, H&E staining, alkaline phosphatase staining. Our results show that BMP2 were able to produce 90-95% healing of critical size defects after six weeks histologically and radiographically. However, SurgiFoam, PRP and L-PRF with or without PGIcNAc were able to close 60% of the original defect. This study supports that BMP2 is more effective for bone regeneration than SurgiFoam, PRP, L-PRF and PGIcNAc.
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A portable platform for stepwise hematopoiesis from human pluripotent stem cells within PET-reinforced collagen sponges / PET繊維補強コラーゲンスポンジを用いた,ヒト多能性幹細胞の段階的な血球分化のための,可搬性のあるプラットフォームSugimine, Yoshinori 24 January 2022 (has links)
京都大学 / 新制・論文博士 / 博士(医学) / 乙第13464号 / 論医博第2251号 / 新制||医||1055(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 金子 新, 教授 江藤 浩之, 教授 髙折 晃史 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM
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