Avaliação da aplicação da terapia fotodinâmica antimicrobiana para o controle e tratamento de infecções envolvendo Candida spp. e Staphylococcus aureus
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Data
2023-03-03
Autores
Barbosa, Elis Rodrigues Oliveira
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Universidade Estadual Paulista (Unesp)
Resumo
Esta dissertação foi dividida em duas publicações que objetivaram: a) realizar uma revisão sistemática da literatura e meta-análise acerca da aplicação clínica e eficácia da TFDa no tratamento de infecções orais envolvendo Candida spp. e b) avaliar o potencial in vitro de frações do extrato de vegetal de Senna spp. como monoterapia na TFDa sobre biofilmes simples e mistos de Candida albicans e Staphylococcus aureus. Publicação 1: Foram realizadas buscas bibliográficas à procura de estudos clínicos randomizados que avaliassem o efeito da TFDa como tratamento de infecções orais envolvendo Candida spp. A revisão seguiu o protocolo PRISMA, e os estudos selecionados foram sintetizados qualitativamente e quantitativamente. O nível de evidência e o risco de viés dos estudos foram avaliados. Meta-análise em rede foi realizada considerando a redução da contagem microbiana no palato e superfície protética. A maioria dos trabalhos investigou o efeito da TFDa na estomatite protética. C. albicans foi a espécie mais prevalente. Todos os estudos incluídos foram classificados como Nível 2 de evidência e apresentaram risco de viés em relação a dados de desfecho ausentes. Quatro artigos foram incluídos na meta-análise em rede. No palato, todos os tratamentos foram eficazes quando comparados ao grupo controle. A TFDa e TFDa+nistatina foram eficazes de forma semelhante, enquanto nistatina ou miconazol apresentaram maior eficácia. A TFDa se mostrou eficaz também na desinfecção das superfícies de próteses. Os resultados sugerem que a aplicação da TFDa é uma abordagem efetiva para o tratamento de infecções orais envolvendo Candida spp. Publicação 2: Extratos brutos de Senna macranthera (SM) e Senna splendida (SS) foram fracionados por cromatografia. As frações tiveram o espectro de absorção de luz verificado e foram caracterizadas quimicamente. Biofilmes simples e mistos de C. albicans e S. aureus foram cultivados durante 48 h. Os grupos testados foram: controle negativo (CN), frações sem irradiação (SM1-, SM2-, SS1-, SS2-), controle de luz (CL), controle de veículo com ou sem luz (DMSO 1%; CV+, CV-, respectivamente) e fração com irradiação (TFDa: SM1+, SM2+, SS1+, SS2+). Cada fração (0,2 mg/ml) foi aplicada nos grupos frações sem irradiação e TFDa e pré-incubadas por 15 min (biofilme simples) ou 20 min (biofilme misto). A irradiação foi feita a 450 nm e 139,5 J/cm2 por 900 s em modo fracionado. Os dados foram analisados nos programas IBM SPSS 20.0 (α=0,05) e R (IC=95%). Foram obtidas 12 frações, dentre as quais quatro apresentaram absorção de luz na região de interesse e foram selecionadas para o estudo. A caracterização fitoquímica evidenciou a presença da classe antraquinonas nas frações ativas. Nos grupos TFDa, todas as frações reduziram a viabilidade microbiana em biofilmes simples e mistos. Na presença de luz, foi observada uma maior redução microbiana nos biofilmes mistos. Apenas a fração SM1 levou a redução > 3 log UFC/mL todos os biofilmes. TFDa mediada por frações dos extratos de Senna spp. demonstraram potencial in vitro para redução de biofilmes simples e mistos de S. aureus e C. albicans.
This Dissertation was divided into two publications that aimed: a) to perform a systematic literature review and meta-analysis on the clinical application and efficacy of aPDT in the treatment of oral infections involving Candida spp. and b) to evaluate the in vitro potential of Senna spp. as photosensitizer in aPDT on single- and dual species biofilms of Candida albicans and Staphylococcus aureus. Publication 1: Electronic searches were performed seeking for randomized clinical trials that evaluated the effect of aPDT as a treatment for oral infections involving Candida spp. The review followed the PRISMA protocol, and the selected studies were synthesized qualitatively and quantitatively. The level of evidence and the risk of bias of each study were assessed. A network meta-analysis was performed considering the reduction of microbial counting on palate and prosthetic surface. Most studies investigated the effect of aPDT on denture stomatitis. C. albicans was the most prevalent species. All included studies were classified as Level 2 of Evidence and had bias risk regarding missing outcome data. Four articles were included in the network meta-analysis. On the palate, all treatments were effective when compared to the control group. aPDT and aPDT+nystatin were similarly effective, while nystatin or miconazole were more effective than aPDT. aPDT was also effective to prosthesis surfaces disinfection. The results suggest that aPDT application represents an effective approach for the treatment of oral infections involving Candida spp. Publication 2: Crude extracts of Senna macranthera (SM) and Senna splendida (SS) were fractionated by the chromatographic method. The fractions had the light absorption spectrum checked and were chemically characterized. Single- and dualspecies biofilms of C. albicans and S. aureus were cultured for 48 h. The following groups were tested: negative control (NC), fractions without light (SM1-, SM2-, SS1-, SS2-), light control (LC), vehicle control with or without light (DMSO 1%; VC+, VC-, respectively) and fractions with irradiation (aPDT: SM1+, SM2+, SS1+, SS2+). Each fraction (0.2 mg/ml) was applied to the fractions without irradiation and aPDT groups and pre-incubated for 15 min (single-species biofilm) or 20 min (dual-species biofilm). The VC+, LC and aPDT groups were irradiated at 450 nm and 139.5 J/cm2 for 900 s in fractionated mode. Data were analyzed using IBM SPSS 20.0 (α=0.05) and R (CI=95%) programs. From the 12 fractions obtained, four showed light absorption between 400 and 450 nm and were selected for the study. Phytochemical characterization showed the presence of anthraquinones class. In aPDT groups, all fractions reduced microbial viability in single- and dual species biofilms. In the presence of light, a greater microbial reduction was observed in dual-species biofilms. Only the SM1 fraction led to > 3 log CFU/mL reduction in all biofilms. aPDT mediated by fractions of Senna spp. demonstrated in vitro potential for the reduction of single- and dual species biofilms of S. aureus and C. albicans.
This Dissertation was divided into two publications that aimed: a) to perform a systematic literature review and meta-analysis on the clinical application and efficacy of aPDT in the treatment of oral infections involving Candida spp. and b) to evaluate the in vitro potential of Senna spp. as photosensitizer in aPDT on single- and dual species biofilms of Candida albicans and Staphylococcus aureus. Publication 1: Electronic searches were performed seeking for randomized clinical trials that evaluated the effect of aPDT as a treatment for oral infections involving Candida spp. The review followed the PRISMA protocol, and the selected studies were synthesized qualitatively and quantitatively. The level of evidence and the risk of bias of each study were assessed. A network meta-analysis was performed considering the reduction of microbial counting on palate and prosthetic surface. Most studies investigated the effect of aPDT on denture stomatitis. C. albicans was the most prevalent species. All included studies were classified as Level 2 of Evidence and had bias risk regarding missing outcome data. Four articles were included in the network meta-analysis. On the palate, all treatments were effective when compared to the control group. aPDT and aPDT+nystatin were similarly effective, while nystatin or miconazole were more effective than aPDT. aPDT was also effective to prosthesis surfaces disinfection. The results suggest that aPDT application represents an effective approach for the treatment of oral infections involving Candida spp. Publication 2: Crude extracts of Senna macranthera (SM) and Senna splendida (SS) were fractionated by the chromatographic method. The fractions had the light absorption spectrum checked and were chemically characterized. Single- and dualspecies biofilms of C. albicans and S. aureus were cultured for 48 h. The following groups were tested: negative control (NC), fractions without light (SM1-, SM2-, SS1-, SS2-), light control (LC), vehicle control with or without light (DMSO 1%; VC+, VC-, respectively) and fractions with irradiation (aPDT: SM1+, SM2+, SS1+, SS2+). Each fraction (0.2 mg/ml) was applied to the fractions without irradiation and aPDT groups and pre-incubated for 15 min (single-species biofilm) or 20 min (dual-species biofilm). The VC+, LC and aPDT groups were irradiated at 450 nm and 139.5 J/cm2 for 900 s in fractionated mode. Data were analyzed using IBM SPSS 20.0 (α=0.05) and R (CI=95%) programs. From the 12 fractions obtained, four showed light absorption between 400 and 450 nm and were selected for the study. Phytochemical characterization showed the presence of anthraquinones class. In aPDT groups, all fractions reduced microbial viability in single- and dual species biofilms. In the presence of light, a greater microbial reduction was observed in dual-species biofilms. Only the SM1 fraction led to > 3 log CFU/mL reduction in all biofilms. aPDT mediated by fractions of Senna spp. demonstrated in vitro potential for the reduction of single- and dual species biofilms of S. aureus and C. albicans.
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Palavras-chave
Fotoquimioterapia, Extratos vegetais, Candida, Extrato de Senna, Photochemotherapy, Plant Extracts, Candida, Senna extract