Efeitos da insulina no tecido cartilagíneo do fêmur de ratos jovens submetidos à lesão térmica
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2021-02-22
Authors
Pereira, Anny Rocha [UNIFESP]
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De Oliveira, Flavia [UNIFESP]
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Trabalho de conclusão de curso de graduação
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Introdução. As lesões térmicas que ocorrem em crianças por derramamento de líquidos quentes estão entre os acidentes mais comuns no mundo. Quando superiores a 40% da extensão corporal são consideradas extensas e, além de respostas locais, levam a repercussões sistêmicas. Objetivo. O objetivo do presente trabalho foi analisar os efeitos do tratamento com insulina em curto e longo prazo após lesão térmica, na morfologia do tecido ósseo e cartilagíneo do fêmur de ratos Wistar jovens. Metodologia. Para isso, foram utilizados 40 ratos Wistar de 21 dias, machos, distribuídos nos grupos: Controle (C), Controle com tratamento diário de insulina (C+I), submetidos à Lesão Térmica por Escaldadura (LTE) e submetidos à Lesão Térmica por Escaldadura com tratamento diário de insulina (LTE+I). Os animais foram eutanasiados após 4 ou 14 dias da lesão ou simulação da mesma. A epífise proximal do fêmur foi submetida à análise histopatológica, morfométrica (mensuração das camadas da cartilagem articular e das camadas da lâmina epifisial, densidade e área do perfil celular dos condrócitos) e de avaliação das fibras colágenas tipos I e III. Realizou-se ainda a análise imunoistoquímica para as proteínas RANX-2 e RANK-L. Foi realizada a análise estatística por meio de análise de variância (ANOVA) com três fatores. Resultados. A lâmina epifisial do fêmur dos animais que sofreram a lesão térmica, em longo prazo apresentou menor tamanho que o controle, independente do tratamento com insulina, essa diminuição se dá nas zonas proliferativa e hipertrófica+calcificada. Os efeitos da insulina só foram verificados em curto prazo após a lesão apenas nas zonas hipertrófica+calcificada. A cartilagem articular sofreu diminuição tanto em longo quanto em curto prazo como efeito da lesão térmica. A insulina diminuiu drasticamente a espessura da cartilagem articular nos grupos C+I, essa diminuição ocorreu em grau mais moderado nos grupos LTE+I também em longo e curto prazo, portanto o efeito modulador da insulina foi diferente na presença da lesão térmica. A interferência da lesão e do tratamento com insulina ocorreu principalmente nas camadas Média+Profunda. Em relação a área dos condrócitos foram observados resultados somente em animais LTE+I de 4 dias que evidenciaram área menor que grupo C+I, demonstrando que ação da insulina foi diferente na presença da lesão térmica. A análise de fibras colágenas da cartilagem articular evidenciou diminuição na coloração de colágeno e comprometimento na malha pantográfica dos animais LTE, após 14 dias da administração de insulina LTE+I apresentou organização semelhante ao grupo C, evidenciando ação positiva da insulina em longo prazo. A avaliação imunoistoquímica para os fatores de regulação osteogênica RUNX-2 e RANK-L não evidenciaram diferença entre os grupos experimentais. No entanto, salienta-se que houve dificuldades na realização da técnica imunoistoquímica. Conclusão. A insulina causou diminuição da cartilagem articular. Já quanto à avaliação do colágeno, a insulina foi capaz de recuperar o padrão em malha pantográfica do colágeno tanto da cartilagem articular quanto da lâmina epifisial.
Introduction. Thermal injuries that occur in children by spilling hot liquids are among the most common accidents in the world. When greater than 40% of the body extension, they are considered extensive and, in addition to local responses, lead to systemic repercussions. Objective. The aim of the present study was to analyze the effects of insulin treatment in the short and long term after thermal injury, on the morphology of bone and cartilage tissue of the femur of young Wistar rats. Methodology. For that, 40 male Wistar rats of 21 days were used, distributed in the groups: Control (C), Control with daily insulin treatment (C + I), submitted to Thermal Injury by Scald (SBI) and submitted to Thermal Injury by Scald with daily insulin treatment (SBI + I). The animals were euthanized after 4 or 14 days of injury or simulation. The proximal epiphysis of the femur was submitted to histopathological, morphometric analysis (measurement of the layers of the articular cartilage and layers of the epiphysial lamina, density and area of the cell profile of the chondrocytes) and evaluation of collagen fibers types I and III. Immunohistochemical analysis was performed for RANX-2 and RANK-L proteins. Statistical analysis was performed using analysis of variance (ANOVA) with three factors. Results. The epiphyseal lamina of the femur of animals that suffered thermal injury, in the long run, was smaller than the control, regardless of insulin treatment, this decrease occurs in the proliferative and hypertrophic + calcified zones. The effects of insulin were only seen in the short term after the injury only in the hypertrophic + calcified areas. Articular cartilage suffered a decrease in both long and short term as an effect of thermal injury. Insulin dramatically decreased the thickness of articular cartilage in groups C + I, this decrease occurred to a more moderate degree in groups SBI + I also in the long and short term, so the insulin modulating effect was different in the presence of thermal injury. Injury and insulin treatment interference occurred mainly in the Middle + Deep layers. Regarding the area of chondrocytes, results were observed only in SBI + I animals of 4 days that showed an area smaller than group C + I, demonstrating that insulin action was different in the presence of thermal injury. The analysis of collagen fibers of the articular cartilage showed a decrease in the collagen staining and impairment in the pantographic mesh of the SBI animals, after 14 days of the administration of insulin SBI + I showed a similar organization to group C, showing positive action of insulin in the long term. The immunohistochemical evaluation for osteogenic regulation factors RUNX-2 and RANK-L did not show any difference between the experimental groups. However, it should be noted that there were difficulties in performing the immunohistochemical technique. Conclusion. Insulin caused a decrease in joint cartilage. As for the evaluation of collagen, insulin was able to recover the pantographic mesh pattern of collagen from both the articular cartilage and the epiphyseal lamina.
Introduction. Thermal injuries that occur in children by spilling hot liquids are among the most common accidents in the world. When greater than 40% of the body extension, they are considered extensive and, in addition to local responses, lead to systemic repercussions. Objective. The aim of the present study was to analyze the effects of insulin treatment in the short and long term after thermal injury, on the morphology of bone and cartilage tissue of the femur of young Wistar rats. Methodology. For that, 40 male Wistar rats of 21 days were used, distributed in the groups: Control (C), Control with daily insulin treatment (C + I), submitted to Thermal Injury by Scald (SBI) and submitted to Thermal Injury by Scald with daily insulin treatment (SBI + I). The animals were euthanized after 4 or 14 days of injury or simulation. The proximal epiphysis of the femur was submitted to histopathological, morphometric analysis (measurement of the layers of the articular cartilage and layers of the epiphysial lamina, density and area of the cell profile of the chondrocytes) and evaluation of collagen fibers types I and III. Immunohistochemical analysis was performed for RANX-2 and RANK-L proteins. Statistical analysis was performed using analysis of variance (ANOVA) with three factors. Results. The epiphyseal lamina of the femur of animals that suffered thermal injury, in the long run, was smaller than the control, regardless of insulin treatment, this decrease occurs in the proliferative and hypertrophic + calcified zones. The effects of insulin were only seen in the short term after the injury only in the hypertrophic + calcified areas. Articular cartilage suffered a decrease in both long and short term as an effect of thermal injury. Insulin dramatically decreased the thickness of articular cartilage in groups C + I, this decrease occurred to a more moderate degree in groups SBI + I also in the long and short term, so the insulin modulating effect was different in the presence of thermal injury. Injury and insulin treatment interference occurred mainly in the Middle + Deep layers. Regarding the area of chondrocytes, results were observed only in SBI + I animals of 4 days that showed an area smaller than group C + I, demonstrating that insulin action was different in the presence of thermal injury. The analysis of collagen fibers of the articular cartilage showed a decrease in the collagen staining and impairment in the pantographic mesh of the SBI animals, after 14 days of the administration of insulin SBI + I showed a similar organization to group C, showing positive action of insulin in the long term. The immunohistochemical evaluation for osteogenic regulation factors RUNX-2 and RANK-L did not show any difference between the experimental groups. However, it should be noted that there were difficulties in performing the immunohistochemical technique. Conclusion. Insulin caused a decrease in joint cartilage. As for the evaluation of collagen, insulin was able to recover the pantographic mesh pattern of collagen from both the articular cartilage and the epiphyseal lamina.
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PEREIRA, Anny Rocha. Efeitos da insulina no tecido cartilagíneo do fêmur de ratos jovens submetidos à lesão térmica. 2021. 40f. Trabalho de conclusão de curso de graduação (Fisioterapia) - Instituto de Saúde e Sociedade, Universidade Federal de São Paulo, Santos, 2021.