Avaliação da terapia fotodinâmica em candida albicans in vitro e in vivo
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Data
2011-07-25
Autores
Costa, Anna Carolina Borges Pereira da [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Os objetivos do presente estudo foram avaliar a ação antifúngica da Terapia Fotodinâmica (TFD) em culturas planctônicas e biofilmes de Candida albicans formados in vitro e em modelo de candidose experimental em camundongos, bem como sua interferência na aderência de C. albicans às células epiteliais bucais humanas in vitro. Foi utilizada cepa padrão de C. albicans (ATCC 18804) para os ensaios. Como fonte de luz foi utilizado o Diodo Emissor de Luz (LED) com emissão de luz verde (532±10 nm) com potência de 90 mW e fotossensibilizador eritrosina nas concentrações variando de 200 a 0,39 μM para os ensaios em cultura plactônica. O biofilme foi formado em placas de 96 poços e tratado com o fotossensibilizador eritrosina na concentração de 400 μM e luz LED. 56 camundongos machos e adultos foram imunossuprimidos e inoculados com suspensões contendo 108 células/mL de C. albicans. Os animais foram submetidos a TFD mediada pelo corante eritrosina (400 μM) e irradiados por LED. Antes e após os tratamentos, foram recuperadas leveduras da cavidade bucal dos animais. As leveduras recuperadas após a TFD e grupo controle foram avaliadas quanto a interferência da TFD na aderência de C. albicans às células do epitélio bucal humano. Em seguida, os animais foram sacrificados e as línguas retiradas para análise macroscópica e histológica. Os biofilmes, línguas e lâminas de aderência foram também analisados por Microscopia Eletrônica de Varredura (MEV). A análise dos dados foi feita por ANOVA e Teste de Tukey, teste t de Student e Kruskal-Wallis (P < 0,05). A TFD aplicada à cultura planctônica de C. albicans foi dose-dependente com redução significativa a partir da menor concentração testada (0,39 μM) e 100% de morte das células a partir de 3,12 μM. A TFD em biofilme formado in vitro reduziu 0,74 log10 de C. albicans com redução de leveduras e hifas...
The aims of the present study were evaluate the antifungal action of Photodynamic Therapy (PDT) on Candida albicans planktonic cultures and biofilms formed in vitro and experimental candidosis model in mice, as well as its interference on C. albicans adherence on humans buccal epithelial cells in vitro. Standard strain of C. albicans (ATCC 18804) was used for assays. Green Light Emitting Diode (LED- 532 ± 10 nm) was used as light source with an output power of 90 mW and erythrosine photosensitizer at concentrations range from 200 to 0.39 μM for the assays on plaktonic cultures. The biofilm was formed on plates of 96- wells and treated with the erythrosine photosensitizer at a concentration of 400 μM and LED light. 56 adults and males mice were immunosuppressed and inoculated with suspensions containing 108 cells/ mL of C. albicans. The animals were submitted to erythrosine dye- mediated PDT (400 μM) and irradiated by LED. Before and after the treatments, yeasts from the animals´ oral cavities were recovered. The yeasts recovered after PDT and control group were evaluated for interference of PDT on C. albicans adherence to humans buccal epithelial cells. Following, the animals were sacrified and their tongues taken away for macroscopic and histological analysis. The biofilms, tongues and adherence slices were also analized by Scanning Electron Microscopy (SEM). The analysis of the dates were done by ANOVA and Tukey test, Student t test and Kruskal-Wallis (P < 0.05). PDT applied on C. albicans planktonic culture was concentrationdependent with significant reduction from minor concentration tested (0.39 μM) and 100% of cells death from 3.12 μM. C. albicans biofilm formed in vitro was reduced 0.74 log10 by PDT with reduction of yeasts and hyphaes verified by SEM. In vivo, 0.73 log10 of C. albicans and 35% adherence on buccal epithelial cells were reduced, but there was not... (Complete abstract click electronic access below)
The aims of the present study were evaluate the antifungal action of Photodynamic Therapy (PDT) on Candida albicans planktonic cultures and biofilms formed in vitro and experimental candidosis model in mice, as well as its interference on C. albicans adherence on humans buccal epithelial cells in vitro. Standard strain of C. albicans (ATCC 18804) was used for assays. Green Light Emitting Diode (LED- 532 ± 10 nm) was used as light source with an output power of 90 mW and erythrosine photosensitizer at concentrations range from 200 to 0.39 μM for the assays on plaktonic cultures. The biofilm was formed on plates of 96- wells and treated with the erythrosine photosensitizer at a concentration of 400 μM and LED light. 56 adults and males mice were immunosuppressed and inoculated with suspensions containing 108 cells/ mL of C. albicans. The animals were submitted to erythrosine dye- mediated PDT (400 μM) and irradiated by LED. Before and after the treatments, yeasts from the animals´ oral cavities were recovered. The yeasts recovered after PDT and control group were evaluated for interference of PDT on C. albicans adherence to humans buccal epithelial cells. Following, the animals were sacrified and their tongues taken away for macroscopic and histological analysis. The biofilms, tongues and adherence slices were also analized by Scanning Electron Microscopy (SEM). The analysis of the dates were done by ANOVA and Tukey test, Student t test and Kruskal-Wallis (P < 0.05). PDT applied on C. albicans planktonic culture was concentrationdependent with significant reduction from minor concentration tested (0.39 μM) and 100% of cells death from 3.12 μM. C. albicans biofilm formed in vitro was reduced 0.74 log10 by PDT with reduction of yeasts and hyphaes verified by SEM. In vivo, 0.73 log10 of C. albicans and 35% adherence on buccal epithelial cells were reduced, but there was not... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Candida albicans, Celulas - Adesão adesão, Candidiase bucal, Terapia fotodinâmica, Eritrosina, Photodynamic therapy, Erythrosine, Oral candidosis
Como citar
COSTA, Anna Carolina Borges Pereira da. Avaliação da terapia fotodinâmica em candida albicans in vitro e in vivo. 2011. 116 f. Dissertação (mestrado ) – Universidade Estadual Paulista, Faculdade de Odontologia de São Jose dos Campos, 2011.