O peróxido de hidrogênio aumenta a eficácia da terapia fotodinâmica em biofilmes de Candida albicans suscetível e resistente ao fluconazol
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Data
2022-03-07
Autores
Sousa, Tábata Viana de
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Universidade Estadual Paulista (Unesp)
Resumo
Candida albicans e a matriz são componentes fundamentais para o desenvolvimento de biofilmes, e os ensaios devem visar ambos os componentes. Nesse contexto, o objetivo deste estudo foi avaliar a eficácia do peróxido de hidrogênio (H2O2) combinado à terapia fotodinâmica antimicrobiana (aPDT), mediada por Photodithazine® (P) e LED (L) sobre biofilme de Candida albicans sensível ao fluconazol (CaS) ATCC 90028 e resistente (CaR) ATCC 96901. Os biofilmes foram cultivados por 48 horas e foram expostos a H2O2 (10mM) por 5 min, seguido pela aplicação de P e L isoladamente ou combinados (H2O2+P+L+, H2O2+P-L+, H2O2, P+L+, PL -, P+L-, P-L+; n=12). Após o tratamento, os biofilmes foram avaliados por unidade formadora de colônia/mL, componentes da matriz extracelular [polissacarídeos solúveis em água (WSP) e em álcali (ASP), proteínas, eDNA], biomassa (total e insolúvel) e proteínas. A Microscopia Eletrônica de Varredura (MEV), foi realizada. Os dados foram analisados por um teste ANOVA de três vias com o teste post-hoc de Bonferroni. A exposição dos biofilmes a 10 mM de H2O2 por 5 minutos seguido de aPDT reduziu as proteínas solúveis e insolúveis da matriz, eDNA, ASP, WSP e o peso seco insolúvel e solúvel dos biofilmes de CaS e de CaR. A MEV revelou redução no número de células do biofilme e dispersão das hifas do grupo H2O2+P+L+ em relação ao P-L- (controle). Em suma, a associação com H2O2 aumentou a eficácia da aPDT e reduziu os componentes da matriz extracelular do biofilme; logo essa associação se mostra promissora para possíveis aplicações in vivo, já que in vitro foi obtido resultados bastante satisfatórios.
Candida albicans and matrix are essential for the development of biofilms, and assays should therefore target both components. The aim of this study was evaluated the effectiveness of the hydrogen peroxide (H2O2) combined with antimicrobial photodynamic therapy (aPDT), mediated by Photodithazine® (P) and LED (L) on biofilms formed by fluconazole susceptible (CaS) and resistant (CaR) Candida albicans strains. Biofilms were grown for 48 hours and were exposed to H2O2 (10mM) for 5 min, followed by application of P and L either singly or combined (H2O2+P+L+, H2O2 +P-L+, H2O2, P+L+, P-L-, P+L-, P-L+; n=12). After the treatment, biofilms were evaluated by colony-forming unit/mL, extracellular matrix components [polysaccharides soluble in water (WSP) and in alkali (ASP), proteins, eDNA,], biomass (total and insoluble) and proteins. Scanning Electron Microscopy (SEM) was performed. The data were analyzed by a three-way ANOVA test with Bonferroni's post-hoc test. In both strains, the treatment of biofilms with 10 mM H2O2 for 5 minutes followed by aPDT reduced soluble and insoluble matrix proteins, eDNA, ASP, WSP and insoluble and soluble dry weight of CaS and CaR biofilms. SEM revealed a reduction in the number of biofilm cells and hyphae dispersion in the H2O2+P+L+ group in relation to the P-L- (control). In short, the association with H2O2 increased the effectiveness of aPDT and reduced the extracellular matrix components of the biofilm; therefore, this association shows promise for possible applications in vivo, since in vitro results were promissory.
Candida albicans and matrix are essential for the development of biofilms, and assays should therefore target both components. The aim of this study was evaluated the effectiveness of the hydrogen peroxide (H2O2) combined with antimicrobial photodynamic therapy (aPDT), mediated by Photodithazine® (P) and LED (L) on biofilms formed by fluconazole susceptible (CaS) and resistant (CaR) Candida albicans strains. Biofilms were grown for 48 hours and were exposed to H2O2 (10mM) for 5 min, followed by application of P and L either singly or combined (H2O2+P+L+, H2O2 +P-L+, H2O2, P+L+, P-L-, P+L-, P-L+; n=12). After the treatment, biofilms were evaluated by colony-forming unit/mL, extracellular matrix components [polysaccharides soluble in water (WSP) and in alkali (ASP), proteins, eDNA,], biomass (total and insoluble) and proteins. Scanning Electron Microscopy (SEM) was performed. The data were analyzed by a three-way ANOVA test with Bonferroni's post-hoc test. In both strains, the treatment of biofilms with 10 mM H2O2 for 5 minutes followed by aPDT reduced soluble and insoluble matrix proteins, eDNA, ASP, WSP and insoluble and soluble dry weight of CaS and CaR biofilms. SEM revealed a reduction in the number of biofilm cells and hyphae dispersion in the H2O2+P+L+ group in relation to the P-L- (control). In short, the association with H2O2 increased the effectiveness of aPDT and reduced the extracellular matrix components of the biofilm; therefore, this association shows promise for possible applications in vivo, since in vitro results were promissory.
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Palavras-chave
Fotoquimioterapia, Candida albicans, Peróxido de hidrogênio, Photochemotherapy, Candida albicans, Hydrogen peroxide