Citotoxicidade da Punica granatum L. (romã) sobre cultura de fibroblastos e de células de linhagem cancerígena
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Data
2009-07-22
Autores
Werkman, Cristina [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Embora diversos estudos já tenham sido realizados sobre a Punica granatum L., seus possíveis efeitos citotóxicos não são bem conhecidos. No presente trabalho foi avaliada a toxicidade da Punica granatum L. (PG) por meio de cultura celular com duas linhagens: fibroblastos humanos de mucosa oral (FLM) e células de carcinoma epidermóide oral humano (KB). As células foram submetidas ao teste de viabilidade celular por 24 horas em placas de 96 poços nas concentrações da PG (1%, 0,5%, 0,25%, 0,125%, 0,062% e 0,031%) e aos testes de citotoxicidade celular em placas de 24 poços, em cinco períodos diferentes (4 horas, 1, 3, 5 e 7 dias) e com quatro diferentes concentrações (1%, 0,5%, 0,25%, 0,062%). Todos os testes foram realizados em triplicata e em três momentos diferentes. Foram adicionados dois grupos, um controle negativo (Tryton 10%) e um controle positivo constituído de meio de cultura acrescido de soro bovino fetal 10% e sem o extrato de PG. A quantificação celular foi realizada pelo método do Resazurin, com avaliação por espectrofotometria óptica (espectrofotômetro UVM340 - Asys Hitech GmbH) e leitura de absorbância dos comprimentos de onda de 570 nm a 600 nm. Os dados obtidos foram submetidos à ANOVA (5%). Os resultados de viabilidade mostraram que nas concentrações de 0.031%; 0,062%; 0,125%; 0,25% e 0,05% o extrato de PG inibiu a viabilidade celular (≥ 40%) principalmente das células KB. O teste de citotoxicidade mostrou que apenas as culturas tratadas com PG a 0,062% exibiram citotoxicidade para as linhagens celulares. A PG a 1% foi tóxica para as células FLM e KB. O extrato de PG inibiu a citotoxicidade celular nas demais concentrações testadas. Estes resultados podem ser relacionados a propriedade anticarcinogênica da Punica granatum L.
Several studies have been conducted on Punica granatum L., but their cytotoxic effects are not well known. The purpose of this study was to evaluate the cytotoxicity of Punica granatum L. extract (PG) in two cell lines: human oral mucosa fibroblasts (FLM) and cells of human oral squamous cell carcinoma (KB). The cells were analized to viability on 24 h using different concentrations of PG (1%, 0.5%, 0.25%, 0.125%, 0.062% and 0.031%) and to cell proliferation in five different periods (4 h, 1, 3, 5 and 7 days) using four different concentrations of PG (1%, 0.5%, 0.25%, 0.062%). All tests were performed in triplicate and in three different times. There were a negative control (Tryton 10%) and a positive control (culture medium plus 10% SBF, without PG extract). Cell quantification was performed by the Resazurin method, with evaluation by optical spectrophotometry (spectrophotometer UVM340 - Asys Hitech GmbH) and measure of absorbance at a wavelength of 570 nm to 600 nm. Data were submitted to ANOVA (5%). The results showed inhibition of cell proliferation in 0,031%, 0.062%, 0.125%, 0.25% and 0.05% concentrations of PG extract (≥ 40%) specialty to KB cells in comparison to FLM cells. PG 1% was toxic to KB and FLM cells. The proliferation test showed cell proliferation only when using PG 0,062%, for both cell lines and inhibited proliferation whith all other PG concentrations. These effects might be related to the anticarcinogenic properties of PG.
Several studies have been conducted on Punica granatum L., but their cytotoxic effects are not well known. The purpose of this study was to evaluate the cytotoxicity of Punica granatum L. extract (PG) in two cell lines: human oral mucosa fibroblasts (FLM) and cells of human oral squamous cell carcinoma (KB). The cells were analized to viability on 24 h using different concentrations of PG (1%, 0.5%, 0.25%, 0.125%, 0.062% and 0.031%) and to cell proliferation in five different periods (4 h, 1, 3, 5 and 7 days) using four different concentrations of PG (1%, 0.5%, 0.25%, 0.062%). All tests were performed in triplicate and in three different times. There were a negative control (Tryton 10%) and a positive control (culture medium plus 10% SBF, without PG extract). Cell quantification was performed by the Resazurin method, with evaluation by optical spectrophotometry (spectrophotometer UVM340 - Asys Hitech GmbH) and measure of absorbance at a wavelength of 570 nm to 600 nm. Data were submitted to ANOVA (5%). The results showed inhibition of cell proliferation in 0,031%, 0.062%, 0.125%, 0.25% and 0.05% concentrations of PG extract (≥ 40%) specialty to KB cells in comparison to FLM cells. PG 1% was toxic to KB and FLM cells. The proliferation test showed cell proliferation only when using PG 0,062%, for both cell lines and inhibited proliferation whith all other PG concentrations. These effects might be related to the anticarcinogenic properties of PG.
Descrição
Palavras-chave
Matéria médica vegetal, Toxicidade - Testes, Medicina alternativa, Punica granatum - Toxicidade, Punica granatum - Cytotoxicity
Como citar
WERKMAN, Cristina. Citotoxicidade da Punica granatum L. (romã) sobre cultura de fibroblastos e de células de linhagem cancerígena. 2009. 115 f. Tese (doutorado) - Universidade Estadual Paulista, Faculdade de Odontologia de São José dos Campos., 2009.