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Avaliação molecular dos mecanismos de bioestimulação fibroblástica utilizando ácido hialurônico e extrato de chá verde (Camellia sinensis)

RESUMO

Objetivo: avaliar a capacidade do extrato de chá verde (Camellia sinensis) e ácido hialurônico em modular a expressão de genes relacionados com a sobrevivência e proliferação de fibroblastos. Material e métodos: após manutenção e expansão, fibroblastos humanos receberam diferentes concentrações (variando de 0% a 100% v/v) de extrato de chá verde e ácido hialurônico para avaliação da adesão celular pela metodologia de cristal violeta. Em seguida, a metodologia RT-qPCR foi aplicada para verificar a expressão dos genes AKT, CDK2, CDK4, VEGF e VEGFR1 frente aos genes controle (ß-ACT, GAPDH e 18S) e calculada pelo método CT. O teste Anova foi aplicado para comparar os resultados (5% significância). Resultados: ambos os compostos impactam na sobrevivência de fibroblastos por meio da ativação do gene AKT. Posteriormente, genes relacionados com a proliferação celular foram avaliados e nossos dados demonstraram que os genes CDK2 e CDK4 foram significantemente expressos em resposta a ambos os compostos analisados. Por fim, o estímulo angiogênico desses compostos foi afirmado pela capacidade de estimular a atividade dos genes VEGF e VEGFR1. Conclusão: em conjunto, os resultados apresentados demonstraram a capacidade do extrato da Camellia sinensis e do ácido hialurônico em promover eventos importantes em fibroblastos, os quais são pré-requisitos em mecanismos de regeneração tecidual. Devido às respostas complementares de ambos os compostos, ensaios de sinergia serão necessários para melhor análise biológica com vista ao desenvolvimento de novos produtos na área odontológica.

Palavras-chave – Regeneração tecidual; Extrato de chá verde (Camellia sinensis); Ácido hialurônico; Fibroblastos; Sinalização celular.

ABSTRACT

Objective: to evaluate the ability of both green tea extract (Camellia sinensis) and hyaluronic acid in modulating the expression of genes related to fibroblast survival and proliferation. Material and methods: after maintenance and expansion, human fibroblasts received di! erent concentrations of these compounds (concentrations ranging from 0% – 100% v/v) of green tea and hyaluronic acid to evaluate cell adhesion by using crystal violet dye. Then, the real-time RTqPCR technique was applied to verify the expression of AKT, CDK2, CDK4, VEGF, VEGFR1 genes against control genes (ß-ACT, GAPDH, 18S) and calculated by the “”CT method. The Anova test was used to compare the results (5% significance). Results: both compounds impact fibroblast survival through the activation of the AKT gene. Subsequently, genes related to cell proliferation were evaluated and our data demonstrated that CDK2 and CDK4 genes were significantly expressed in response to both analyzed compounds. Finally, the angiogenic stimulus of these compounds was confirmed by the ability to stimulate the activity of the VEGF and VEGFR1 genes. Conclusion: together, the results presented demonstrate the ability of Camellia sinensis extract and hyaluronic acid to promote important events in fibroblasts, which are prerequisites in tissue regeneration mechanisms. Due to the complementary responses of both compounds, synergy assays will be necessary for a better biological analysis with a view to the development of new products in the dental field.

Key words – Tissue regeneration; Green tea (Camellia sinensis) extract; Hyaluronic acid; Fibroblasts; Cell signaling.

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