Avaliação antifúngica e citotoxicidade do ácido peracético e extrato de Camellia sinensis (chá verde) em Candida spp
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Data
2011-06-12
Autores
Majewski, Marta [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O objetivo deste trabalho foi avaliar in vitro a atividade antifúngica e a citotoxicidade do ácido peracético e extrato vegetal de Camellia sinensis a (chá verde) e também avaliar o efeito antifúngico da anfotericina B e fluconazol em 22 cepas de Candida spp. isoladas da cavidade bucal de pacientes HIV positivos. Para determinação da concentração fungicida mínima e concentração inibitória mínima (CFM e CIM) em células planctônicas, foram realizadas diluições seriadas do extrato e do ácido peracético em placas de 96 poços em caldo RPMI tamponado, em suspensões padronizadas de cada cepa de Candida (106 células/mL). Após determinação da concentração do extrato e do ácido peracético no CIM e CFM, foi preparado o biofilme de cada cepa em placas de 24 poços utilizando-se corpos-de-prova de resina acrílica (n=10), as quais foi adicionado 0,1 mL da suspensão de Candida e 2mL caldo BHI sacarosado. As placas foram incubadas e mantidas a 37° por 5 dias. Após a incubação, os corpos-de-prova foram lavados em solução fisiológica e colocados em imersão por 5 minutos no ácido peracético ou no extrato vegetal. Os corpos-de-prova foram sonicados e realizadas diluições seriadas, 0,1 ml de cada diluição foi semeada em placas de ágar Sabouraud em duplicata e incubadas em estufa bacteriológica (37ºC/48 h). Posteriormente foi realizada a contagem das unidades formadoras de colônia (UFC/mL). Os dados foram avaliados estatisticamente utilizando-se teste de Mann Whitney (p<0,05) para o biofilme, observação visual para CIM e CFM, Tukey e Anova para citotoxicidade. Foi avaliada a citotoxicidade do ácido peracético e extrato de chá verde em macrófagos de camundongos (RAW 264.7), para verificar a viabilidade celular...
The objective of this study was to evaluate the in vitro antifungal activity and cytotoxicity of peracetic acid and plant extract from the Camellia sinensis (green tea) and also to evaluate the antifungal effect of amphotericin B and fluconazole in 22 strains of Candida spp. isolated from the oral cavity of HIV positive patients. To determine the minimum fungicidal concentration and minimum inhibitory concentration (CFM and CIM) in planktonic cells, were performed serial dilutions of the extract and peracetic acid in 96-well plates in RPMI broth buffered in standardized suspensions of each strain of Candida (106 cells / mL). After determining the concentration of the extract and peracetic acid in CIM and CFM, the biofilm was prepared for each strain in 24-well plates using body-of-proof acrylic resin (n = 10), which was added 0, 1 mL of the suspension of Candida sucrose and 2 ml BHI broth. The plates were incubated at 37 ° and maintained for 5 days. After incubation, the bodies of the test piece was washed in saline and placed for 5 minutes immersion in peracetic acid or the plant extract. The bodies of the test piece were sonicated and serial dilutions performed, 0.1 ml of each dilution was seeded on Sabouraud agar plates in duplicate and incubated in a bacteriological incubator (37 º C/48 h). Was subsequently performed the counting of colony forming units (CFU / mL). The data were statistically analyzed using Mann Whitney test (p <0.05) for the biofilm, visual observation for CIM and CFM, ANOVA and Tukey for cytotoxicity. We evaluated the cytotoxicity of peracetic acid and green tea extract in mouse macrophages (RAW 264.7) to verify the cell viability of these substances. The results showed that there was feasibility of peracetic acid and green tea extract in the cells analyzed. The antifungal agents have... (Complete abstract click electronic access below)
The objective of this study was to evaluate the in vitro antifungal activity and cytotoxicity of peracetic acid and plant extract from the Camellia sinensis (green tea) and also to evaluate the antifungal effect of amphotericin B and fluconazole in 22 strains of Candida spp. isolated from the oral cavity of HIV positive patients. To determine the minimum fungicidal concentration and minimum inhibitory concentration (CFM and CIM) in planktonic cells, were performed serial dilutions of the extract and peracetic acid in 96-well plates in RPMI broth buffered in standardized suspensions of each strain of Candida (106 cells / mL). After determining the concentration of the extract and peracetic acid in CIM and CFM, the biofilm was prepared for each strain in 24-well plates using body-of-proof acrylic resin (n = 10), which was added 0, 1 mL of the suspension of Candida sucrose and 2 ml BHI broth. The plates were incubated at 37 ° and maintained for 5 days. After incubation, the bodies of the test piece was washed in saline and placed for 5 minutes immersion in peracetic acid or the plant extract. The bodies of the test piece were sonicated and serial dilutions performed, 0.1 ml of each dilution was seeded on Sabouraud agar plates in duplicate and incubated in a bacteriological incubator (37 º C/48 h). Was subsequently performed the counting of colony forming units (CFU / mL). The data were statistically analyzed using Mann Whitney test (p <0.05) for the biofilm, visual observation for CIM and CFM, ANOVA and Tukey for cytotoxicity. We evaluated the cytotoxicity of peracetic acid and green tea extract in mouse macrophages (RAW 264.7) to verify the cell viability of these substances. The results showed that there was feasibility of peracetic acid and green tea extract in the cells analyzed. The antifungal agents have... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Candida spp, Fitoterápicos, Ácido peracético - Citotoxicidade, Camellia sinensis - Citotoxicidade
Como citar
MAJEWSKI, Marta. Avaliação antifúngica e citotoxicidade do ácido peracético e extrato de Camellia sinensis (chá verde) em Candida spp. 2011. 75 f. Tese (doutorado) – Universidade Estadual Paulista, Faculdade de Odontologia de São José dos Campos, 2011.