Citotoxicidade do ácido peracético: avaliação metabólica, estrutural e de morte em fibroblastos L929
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Data
2016-03-17
Autores
Viola, Kennia Scapin [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A solução de ácido peracético (AP) vem sendo citada na literatura odontológica como uma possível solução irrigadora de canais radiculares. Ele apresenta a vantagem de reunir em um só produto atividade antibacteriana e capacidade de remoção de tecido orgânico e de smear layer. Para a seleção da solução irrigadora de canais radiculares deve-se levar em consideração não somente os seus benefícios terapêuticos, mas também os possíveis efeitos citotóxicos, uma vez que ela pode entrar em contato com os tecidos periapicais. O objetivo deste estudo foi investigar a citotoxicidade e o mecanismo de agressão celular do AP em comparação com o hipoclorito de sódio (NaOCl) sobre fibroblastos L929 por meio de avaliação metabólica, estrutural, ultraestrutural e de morte celular (necrose e/ou apoptose). As células foram incubadas com diferentes concentrações do AP a 1% e do NaOCl a 2,5%, por 10 minutos. Após este período, foram avaliados o metabolismo celular por meio do ensaio do metiltetrazólio (MTT), a morfologia externa das células em microscópio eletrônico de varredura, a ultraestrutura em microscópio eletrônico de transmissão, o citoesqueleto por meio de marcação para actina e α- tubulina, o tipo de morte celular (necrose ou apoptose) por citometria de fluxo. Os resultados foram analisados empregando ANOVA de dois fatores e pós-teste de Bonferroni (α=0.05). Observou-se que o AP apresentou maior citotoxicidade do que o NaOCl. As alterações no metabolismo, na viabilidade, nos filamentos de actina e nos microtúbulos, na morfologia externa e na ultraestrtura das células foram observadas em concetrações menores do AP quando comparado com o NaOCl. A morte celular provocada tanto para o AP quanto para o NaOCl ocorreu predominantemente por necrose. Conclui-se que, embora apresentem o mesmo mecanismo de citotoxicidade, o AP 1% é mais citotóxico que o NaOCl a 2,5%.
The solution peracetic acid (PA) has been reported in the dental literature as a possible root canal irrigating solution. It has the advantage of attending together in one product antibacterial activity and ability to remove of organic tissue and smear layer. For selection of root canal irrigating solution it should take into account not only its therapeutic benefits, but also the possible cytotoxic effects, since it can contact the periapical tissues. The aim of this study was to evaluate the aggression mechanism of 1% AP compared with 2.5% sodium hypochlorite (NaOCl) on L929 fibroblasts through metabolic evaluation, structural, ultrastructural and cell death (necrosis and / or apoptosis). The cells were incubated with different dilutions of the 1% AP and 2.5% NaOCl for 10 minutes. After this period, it was evaluated the cell metabolism by methyl tetrazolium (MTT) assay, the cell external morphology in scanning electron microscope, the ultrastructure in transmission electron microscope, the cytoskeleton by marking to actin and α-tubulin, the type of cell death (necrosis or apoptosis) by flow cytometry. The results were analyzed using two-way ANOVA and Bonferroni post-test (α = 0.05). It was observed that the AP showed higher cytotoxicity than NaOCl. Changes in metabolism, viability, in actin filaments and microtubules, surface morphology and ultraestrtura cells were observed at lower dilutions AP when compared to NaOCl. Cell death caused for both the AP and for NaOCl occurred predominantly by necrosis. It was concluded, though they have the same cytotoxicity mechanism, 1% AP is more cytotoxic than the 2.5% NaOCl.
The solution peracetic acid (PA) has been reported in the dental literature as a possible root canal irrigating solution. It has the advantage of attending together in one product antibacterial activity and ability to remove of organic tissue and smear layer. For selection of root canal irrigating solution it should take into account not only its therapeutic benefits, but also the possible cytotoxic effects, since it can contact the periapical tissues. The aim of this study was to evaluate the aggression mechanism of 1% AP compared with 2.5% sodium hypochlorite (NaOCl) on L929 fibroblasts through metabolic evaluation, structural, ultrastructural and cell death (necrosis and / or apoptosis). The cells were incubated with different dilutions of the 1% AP and 2.5% NaOCl for 10 minutes. After this period, it was evaluated the cell metabolism by methyl tetrazolium (MTT) assay, the cell external morphology in scanning electron microscope, the ultrastructure in transmission electron microscope, the cytoskeleton by marking to actin and α-tubulin, the type of cell death (necrosis or apoptosis) by flow cytometry. The results were analyzed using two-way ANOVA and Bonferroni post-test (α = 0.05). It was observed that the AP showed higher cytotoxicity than NaOCl. Changes in metabolism, viability, in actin filaments and microtubules, surface morphology and ultraestrtura cells were observed at lower dilutions AP when compared to NaOCl. Cell death caused for both the AP and for NaOCl occurred predominantly by necrosis. It was concluded, though they have the same cytotoxicity mechanism, 1% AP is more cytotoxic than the 2.5% NaOCl.
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Palavras-chave
Ácido peracético, Endodontia, Hipoclorito de sódio, Toxicidade, Fibroblastos, Peracetic acid, Endodontics, Sodium hypochlorite, Toxicity, Fibroblasts