Viabilidade e morte celular em queimaduras In vitro

• Ano de defesa: 2018
• Autor(a) principal: Fernandes, Ana Carolina Morais [UNIFESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo (UNIFESP)
• Palavras-chave em Português: Queimaduras; Linhagem celular; Células NIH 3t3; Fibroblastos; Morte celular
• Link de acesso: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=6371073; https://repositorio.unifesp.br/handle/11600/52530

Introduction: Burns are responsible for pathophysiological, cellular and molecular changes and represent a severe form of trauma. There are few in vitro studies that allow the analysis of viability and cell death in thermal burns. Objective: To develop an experimental in vitro burn model capable of evaluating the activity of skin cells such as NIH-3T3 fibroblast line cells under thermal stress conditions for specific temperature values. Characterizing the cellular morphology and the process of cell death by necrosis and / or apoptosis associated with thermal burn, impossible to be evaluated for skin cells in vivo. Methods: 1) NIH-3T3 fibroblast line cells were used, which were subjected to thermal burn for 30s at specified temperatures; 2) The effects of thermal injury on cell viability were determined by: MTT assay, flow cytometry and immunohistochemistry; 3) statistical analysis was performed by the ANOVA variance test, for p <0.05, considering the mean and standard error of quadriplicates, n=4. Results: the MTT assay determined two graphical curves (two phases) with an inflection point for the delta temperature of 15 ° C; flow cytometric analysis indicated that with the increase of the delta temperature there was progressively increase in apoptosis followed by progressive increase in necrosis; the immunohistochemistry evidenced that with the increase in temperature there was a decrease in number of cells and number of mitosis, changes in membrane structure and cytoskeleton (formation of vesicles), with pycnotic and irregular nuclei. Conclusion: With the increase of thermal injury temperature there were significant changes in cellular activity and viability; drastic changes in morphology, with cellular lysis and progressive increase of apoptosis and necrosis of NIH 3T3 cells.