Atividade antimicrobiana e citotoxicidade dos extratos glicólicos de Pfaffia paniculata K. E Juglans regia L.
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2016-12-19
Autores
Ramos, Lucas de Paula [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O objetivo do trabalho foi investigar se os extratos de Pfaffia paniculata K. e Juglans regia L. possuem ação antifúngica, antibacteriana e toxicidade celular, com testes in vitro. Para os testes antifúngicos foram utilizadas cepas ATCC de Candida spp., e para os testes antibacterianos foram utilizadas cepas ATCC de Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans e Pseudomonas aeruginosa. Para a atividade antimicrobiana primeiramente foram determinados os valores da Concentração Inibitória Mínima (CIM) e da Concentração Microbicida Mínima (CMM) dos extratos pelo método de microdiluição em caldo, segundo Clinical and Laboratory Standards Institute (CLSI). Os micro-organismos que apresentaram CMM foram selecionados para os testes em biofilme, no qual foi preparado em fundo de placa com 96 poços, por 48 h. Após os biofilmes foram tratados por 5 min. utilizando as concentrações de 200, 100 e 50 mg dos extratos. Para mensuração da biomassa foi utilizado o teste de Cristal violeta (CV), e para avaliar a atividade metabólica foi utilizado o teste de MTT. A citotoxicidade foi avaliada sobre fibroblastos gengivais humanos (FMM-1) utilizando os mesmos parâmetros de tratamento utilizados para os testes em biofilmes. Foram avaliadas a viabilidade celular pelos testes de MTT, vermelho neutro e cristal violeta. Os dados obtiveram distribuição normal e foram analisados por ANOVA e teste de Tukey, com significância de 5% (p<0.05%). O extrato de P. paniculata demostrou ação antifúngica em biofilmes, com reduções médias de 29,4 e 42,7% nos testes de CV e MTT. Já a ação antibacteriana foi restrita a S. mutans e P. aeruginosa com reduções médias de 15,7 e 28,6% nos respectivos testes. O extrato de J. regia também demostrou ação antifúngica com redução média de 22,2% na biomassa e 31,4% na atividade metabólica. A ação antimicrobiana ficou restrita a P. aeruginosa com reduções médias de 17,7 e 15,6%, indicados pelos testes de CV e MTT. Quanto a citotoxicidade, a média entre os três testes realizados, indicou que após exposição ao extrato de P. paniculata 58,8% das células continuaram viáveis e para J. regia a viabilidade foi de 65,1%. Conclui-se queo extrato de P. paniculata demostrou ação antifúngica sobre todas as cepas de Candida spp. testadas e demostrou ação antibacteriana para P. aeruginosa e S. mutans. As concentrações de 200, 100 e 50 mg do extrato demostraram ser citotóxicas conforme nova diretriz de toxicidade. J. regia demostrou ação antifúngica sobre todas as cepas de Candida spp. testadas e demostrou ação antibacteriana sobre P. aeruginosa. Apenas a concentração de 200 mg do extrato se mostrou tóxica a FMM-1.
The aim of this study was to investigate whether extracts of Pfaffia paniculata K. and Juglans regia L. have antifungal, antibacterial and cellular toxicity, with in vitro tests. ATCC strains of Candida spp. Were used for antifungal tests, and ATCC strains of Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans and Pseudomonas aeruginosa were used for the antibacterial tests. For the antimicrobial activity, the values of the Minimum Inhibitory Concentration (MIC) and the Minimal Microbicidal Concentration (CMM) of the extracts were determined by the microdilution method in broth, according to Clinical and Laboratory Standards Institute (CLSI). The microorganisms that presented CMM were selected for the biofilm tests, in which it was prepared on a 96-well plate bottom for 48 h. After the biofilms were treated for 5 min. Using the concentrations of 200, 100 and 50 mg of the extracts. To measure the biomass, the Violet Crystal test (CV) was used, and the MTT test was used to evaluate the metabolic activity. Cytotoxicity was assessed on human gingival fibroblasts (FMM-1) using the same treatment parameters used for biofilm tests. Cell viability was evaluated by the MTT, neutral red and violet crystal tests. The data obtained normal distribution and were analyzed by ANOVA and Tukey test, with significance of 5%. The extract of P. paniculata showed antifungal action in biofilms, with average reductions of 29.4 and 42.7% in CV and MTT tests; The antibacterial action was restricted to S. mutans and P. aeruginosa with mean reductions of 15.7 and 28.6% in the respective tests. The extract of J. regia also demonstrated antifungal action with an average reduction of 22.2% in biomass and 31.4% in metabolic activity. The antimicrobial action was restricted to P. aeruginosa with mean reductions of 17.7 and 15.6%, indicated by CV and MTT tests. As for cytotoxicity, the mean of the three tests carried out indicated that after exposure to P. paniculata extract 58.8% of the cells remained viable and for viability the viability was 65.1%. In conclusion the extract of P. paniculata showed antifungal action on all strains of Candida spp. Tested and demonstrated antibacterial action for P. aeruginosa and S. mutans. The concentrations of 200, 100 and 50 mg of the extract proved to be cytotoxic according to the new toxicity guideline. J. regia demonstrated antifungal action on all strains of Candida spp. Tested and demonstrated antibacterial action on P. aeruginosa. Only the 200 mg concentration of the extract was shown to be toxic to FMM-1.
The aim of this study was to investigate whether extracts of Pfaffia paniculata K. and Juglans regia L. have antifungal, antibacterial and cellular toxicity, with in vitro tests. ATCC strains of Candida spp. Were used for antifungal tests, and ATCC strains of Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans and Pseudomonas aeruginosa were used for the antibacterial tests. For the antimicrobial activity, the values of the Minimum Inhibitory Concentration (MIC) and the Minimal Microbicidal Concentration (CMM) of the extracts were determined by the microdilution method in broth, according to Clinical and Laboratory Standards Institute (CLSI). The microorganisms that presented CMM were selected for the biofilm tests, in which it was prepared on a 96-well plate bottom for 48 h. After the biofilms were treated for 5 min. Using the concentrations of 200, 100 and 50 mg of the extracts. To measure the biomass, the Violet Crystal test (CV) was used, and the MTT test was used to evaluate the metabolic activity. Cytotoxicity was assessed on human gingival fibroblasts (FMM-1) using the same treatment parameters used for biofilm tests. Cell viability was evaluated by the MTT, neutral red and violet crystal tests. The data obtained normal distribution and were analyzed by ANOVA and Tukey test, with significance of 5%. The extract of P. paniculata showed antifungal action in biofilms, with average reductions of 29.4 and 42.7% in CV and MTT tests; The antibacterial action was restricted to S. mutans and P. aeruginosa with mean reductions of 15.7 and 28.6% in the respective tests. The extract of J. regia also demonstrated antifungal action with an average reduction of 22.2% in biomass and 31.4% in metabolic activity. The antimicrobial action was restricted to P. aeruginosa with mean reductions of 17.7 and 15.6%, indicated by CV and MTT tests. As for cytotoxicity, the mean of the three tests carried out indicated that after exposure to P. paniculata extract 58.8% of the cells remained viable and for viability the viability was 65.1%. In conclusion the extract of P. paniculata showed antifungal action on all strains of Candida spp. Tested and demonstrated antibacterial action for P. aeruginosa and S. mutans. The concentrations of 200, 100 and 50 mg of the extract proved to be cytotoxic according to the new toxicity guideline. J. regia demonstrated antifungal action on all strains of Candida spp. Tested and demonstrated antibacterial action on P. aeruginosa. Only the 200 mg concentration of the extract was shown to be toxic to FMM-1.
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Palavras-chave
Pfaffia paniculata, Juglans regia, Fitoterapia, Antimicrobianos, Phytotherapy, Anti-infective agents