Análise da expressão dos fatores de crescimento FGF, VEGF e TGF associados ao reparo dérmico estimulado pelo látex natural (Hevea brasiliensis) e seu efeito na taxa de pulsação em Lumbriculus variegatus
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Data
2022-12-16
Autores
Paulino, Cristiane Garcia [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Estudos recentes revelam a eficácia do uso do látex natural, extraído da seringueira Hevea brasiliensis, como agente cicatrizante e como uma matriz carreadora de diversos produtos com efeitos farmacológicos. Por ser um evento sistêmico, a cicatrização abrange uma gama de fatores que precisam interagir entre si para que haja uma evolução de forma eficiente. Tendo em vista a capacidade angiogênica e cicatrizante do látex, esta pesquisa investigou através de análises moleculares a expressão dos fatores de crescimento FGF, VEGF e TGF presentes no látex, em cultura de fibroblastos 3T3. Dentre os fatores de crescimento que influenciam o crescimento dérmico, podemos destacar o fator de crescimento do endotélio vascular (VEGF), fator de crescimento transformador beta (TGF-β) e o fator de crescimento fibroblástico (FGF), que estimulam a proliferação celular, a produção de colágeno e de outros elementos da matriz celular, favorecendo o processo de cicatrização. Conforme as análises de FTIR, os componentes identificados presentes na membrana de látex, na fração F1 e no soro foram identificados pertencentes ao isopreno, principal constituinte do látex, como para os grupos funcionais. A amostra do soro apresentou maior discrepância evidenciando que a parte proteica pode estar agindo como indutor de angiogênese. O ensaio hemolítico das amostras, fração F1, soro e a membrana de látex não apresentaram um alto grau de hemólise significativa no período observado, demonstrando assim ser um biomaterial hemocompatível. Nas condições experimentais utilizadas, evidenciou-se baixo teor de fenóis e capacidade antioxidante da fração F1, soro e membrana de látex. Os resultados obtidos nos ensaios com os Blackworms indicam que as frações F1, soro e membrana de látex, nas concentrações testadas, tiveram efeito no sistema circulatório alterando a taxa de pulsação dos Blackworms. A fração F1 e o soro do látex, na concentração de 0,01%, no cultivo de fibroblastos e macrófagos apresentaram melhor viabilidade celular no período de 72h de incubação. A membrana de látex apresentou viabilidade celular superior em todos os períodos de incubação quando comparados ao controle, porém seus resultados foram mais satisfatórios no intervalo de 72h. Os resultados encontrados da análise de expressão gênica confirmaram que houve aumento significativo nas expressões dos genes VEGF, FGF e TGF na membrana e nas frações F1 e soro do látex quando comparadas ao controle, evidenciando o potêncial angiogênico dessas amostras. Este mecanismo de Engenharia Tecidual pode ser um dos procedimentos para regeneração dérmica no futuro.
Studies prove the effectiveness of using natural latex, extracted from the rubber tree Hevea brasiliensis, as a healing agent and as a carrier matrix for various products with pharmacological effects. As it is a systemic event, healing encompasses a range of factors that need to interact with each other so that there is an efficient evolution. In view of the angiogenic and healing capacity of latex, the purpose of this research will be to investigate, through molecular analysis, the expression of growth factors FGF, VEGF and TGF present in latex, in culture of 3T3 fibroblasts. Among the growth factors that influence dermal growth, we can highlight the vascular endothelial growth factor (VEGF), transforming growth factor beta (TGF-β) and fibroblastic growth factor (FGF), which stimulate cell proliferation, the production of collagen and other elements of the cellular matrix, favoring the healing process. According to the FTIR analyses, the identified components present in the latex membrane, in the F1 fraction and in the serum were identified belonging to isoprene, the main constituent of latex, as for the functional groups. The serum sample showed greater discrepancy, showing that the protein part may be acting as an angiogenesis inducer. The hemolytic assay of the samples, F1 fraction, serum and the latex membrane did not show a significant high degree of hemolysis in the observed period, thus demonstrating that it is a hemocompatible biomaterial. Under the experimental conditions used, a low phenol content and antioxidant capacity were found in the F1 fraction, serum and latex membrane. The results obtained in the tests with the Blackworms indicate that the F1 fractions, serum and latex membrane, in the tested concentrations, had an effect on the circulatory system, altering the pulsation rate of the Blackworms. The F1 fraction and the latex serum, at a concentration of 0.01%, in the culture of fibroblasts and macrophages showed better cell viability in the period of 72 hours of incubation. The latex membrane showed superior cell viability in all incubation periods when compared to the control, but its results were more satisfactory in the 72h interval. The results found from the gene expression analysis confirmed that there was a significant increase in the expressions of the VEGF, FGF and TGF genes in the membrane and in the F1 fractions and latex serum when compared to the control, evidencing the angiogenic potential of these samples. This Tissue Engineering mechanism may be one of the procedures for dermal regeneration in the future.
Studies prove the effectiveness of using natural latex, extracted from the rubber tree Hevea brasiliensis, as a healing agent and as a carrier matrix for various products with pharmacological effects. As it is a systemic event, healing encompasses a range of factors that need to interact with each other so that there is an efficient evolution. In view of the angiogenic and healing capacity of latex, the purpose of this research will be to investigate, through molecular analysis, the expression of growth factors FGF, VEGF and TGF present in latex, in culture of 3T3 fibroblasts. Among the growth factors that influence dermal growth, we can highlight the vascular endothelial growth factor (VEGF), transforming growth factor beta (TGF-β) and fibroblastic growth factor (FGF), which stimulate cell proliferation, the production of collagen and other elements of the cellular matrix, favoring the healing process. According to the FTIR analyses, the identified components present in the latex membrane, in the F1 fraction and in the serum were identified belonging to isoprene, the main constituent of latex, as for the functional groups. The serum sample showed greater discrepancy, showing that the protein part may be acting as an angiogenesis inducer. The hemolytic assay of the samples, F1 fraction, serum and the latex membrane did not show a significant high degree of hemolysis in the observed period, thus demonstrating that it is a hemocompatible biomaterial. Under the experimental conditions used, a low phenol content and antioxidant capacity were found in the F1 fraction, serum and latex membrane. The results obtained in the tests with the Blackworms indicate that the F1 fractions, serum and latex membrane, in the tested concentrations, had an effect on the circulatory system, altering the pulsation rate of the Blackworms. The F1 fraction and the latex serum, at a concentration of 0.01%, in the culture of fibroblasts and macrophages showed better cell viability in the period of 72 hours of incubation. The latex membrane showed superior cell viability in all incubation periods when compared to the control, but its results were more satisfactory in the 72h interval. The results found from the gene expression analysis confirmed that there was a significant increase in the expressions of the VEGF, FGF and TGF genes in the membrane and in the F1 fractions and latex serum when compared to the control, evidencing the angiogenic potential of these samples. This Tissue Engineering mechanism may be one of the procedures for dermal regeneration in the future.
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Palavras-chave
Látex, Cicatrização, Citocinas, Expressão gênica, Oligochaeta