Biofilmes heterotípicos formados por C.albicans e espécies não albicans: estudo da interação microbiana e da sensibilidade à terapia fotodinâmica
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Data
2012-07-23
Autores
Piva, Elisabete [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A maioria das infecções por Candida estão relacionadas com biofilmes microbianos, que muitas vezes são formados por associações de diferentes espécies. O objetivo deste estudo foi avaliar as interações entre as diferentes espécies de Candida em biofilmes formados in vitro e verificar a sensibilidade dos biofilmes monotípicos de C. albicans e heterotípicos à terapia fotodinâmica (PDT). Foram estudadas cepas clínicas de C. albicans, C. tropicalis, C. glabrata e C. krusei. A partir de suspensões padronizadas de cada cepa (107 células/mL), biofilmes monotípicos e heterotípicos (formados por C. albicans e espécies não albicans) foram formados em placas de microtitulação de 96 poços durante 48 horas. A seguir, os biofilmes monotípicos de C. albicans e heterotípicos foram submetidos à PDT com fotossensibilizador eritrosina associado ao diodo emissor de luz (LED) verde. Os biofilmes foram analisados por meio da contagem de unidades formadoras de colônia (UFC/mL) em HiCrome Candida por 48h e pela viabilidade celular através do ensaio colorimétrico XTT (2 metoxi 4 nitro 5 sulfofenil 5 fenilalanina carbonil 2H tetrazolium hidróxido). Os resultados obtidos foram submetidos à ANOVA e teste de Tukey. Tanto na contagem de UFC/mL quanto no ensaio colorimétrico XTT, foi observado que o crescimento de C. albicans em biofilme monotípico e heterotípicos com C. tropicalis e C. glabrata foi semelhante. Entretanto, foi inibido em biofilme heterotípico com C. krusei. Foi demonstrado também que a PDT reduziu a viabilidade celular e a contagem de UFC/mL tanto dos biofilmes monotípicos quanto heterotípicos. Concluiu-se que C. albicans estabeleceu relações favoráveis com C. tropicalis e C. glabrata e relações antagônicas com C. krusei em biofilmes heterotípicos. Além disso, os biofilmes monotípicos e heterotípicos foram sensíveis à PDT mediada por LED e eritrosina
Most Candida infections are related to microbial biofilm, which are often formed by the association of different species. The objective of this study was to evaluate the interactions between different species of Candida biofilms in vitro and investigate the sensitivity of C. albicans monotypic biofilms and heterotypic biofilms to photodynamic therapy (PDT). From standardized suspensions of each strain (107 cells/ml), monotypic and heterotypic biofilms (formed by C. albicans and non-albicans species) were developed on flat-bottom 96-well microtiter plates for 48 hours. After that, C. albicans monotypic biofilms and heterotypic biofilms were subjected to PDT with photosensitizer erythrosine associated with green light emitting diode (LED). Biofilms were analysed by counting colony-forming units (CFU/m)l in Candida HiCrome for 48h and by cell viability using the XTT (2 methoxy 4 nitro 5 sulfophenyl 5 phenylamino carbonyl 2H tetrazolium hydroxide) colorimetric assay. The results were subjected to ANOVA and Tukey test. Both in the counting of CFU/ml as in the XTT colorimetric assay, it was observed that the growth of C. albicans in monotypic and heterotypic biofilm with C. glabrata and C. tropicalis was similar. However, it was inhibited in heterotypic biofilm with C. krusei. It was also demonstrated that PDT reduced cell viability and the number of CFU/mL of both monotypic and heterotypic biofilms. It was concluded that C albicans had favorable interaction with C. glabrata and C. tropicalis and antagonistic interaction with C.krusei in heterotypic biofilms. In addition, both of monotypic and heterotypic biofilms were sensitive to LED- and erythrosine-mediated Photodynamic Therapy
Most Candida infections are related to microbial biofilm, which are often formed by the association of different species. The objective of this study was to evaluate the interactions between different species of Candida biofilms in vitro and investigate the sensitivity of C. albicans monotypic biofilms and heterotypic biofilms to photodynamic therapy (PDT). From standardized suspensions of each strain (107 cells/ml), monotypic and heterotypic biofilms (formed by C. albicans and non-albicans species) were developed on flat-bottom 96-well microtiter plates for 48 hours. After that, C. albicans monotypic biofilms and heterotypic biofilms were subjected to PDT with photosensitizer erythrosine associated with green light emitting diode (LED). Biofilms were analysed by counting colony-forming units (CFU/m)l in Candida HiCrome for 48h and by cell viability using the XTT (2 methoxy 4 nitro 5 sulfophenyl 5 phenylamino carbonyl 2H tetrazolium hydroxide) colorimetric assay. The results were subjected to ANOVA and Tukey test. Both in the counting of CFU/ml as in the XTT colorimetric assay, it was observed that the growth of C. albicans in monotypic and heterotypic biofilm with C. glabrata and C. tropicalis was similar. However, it was inhibited in heterotypic biofilm with C. krusei. It was also demonstrated that PDT reduced cell viability and the number of CFU/mL of both monotypic and heterotypic biofilms. It was concluded that C albicans had favorable interaction with C. glabrata and C. tropicalis and antagonistic interaction with C.krusei in heterotypic biofilms. In addition, both of monotypic and heterotypic biofilms were sensitive to LED- and erythrosine-mediated Photodynamic Therapy
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Palavras-chave
Candida, Biofilme, Eritrosina
Como citar
PIVA, Elisabete. Biofilmes heterotípicos formados por C.albicans e espécies não albicans: estudo da interação microbiana e da sensibilidade à terapia fotodinâmica. 2012. 80 f. Dissertação (mestrado) – Universidadde Estadual Paulista. Faculdade de Odontologia de São José dos Campos, 2012.