Avaliação da terapia fotodinâmica contra biofilmes acneicos polimicrobianos: um estudo pré-clínico
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Data
2022-05-25
Autores
De Annunzio, Sarah Raquel
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A acne vulgar é uma das doenças dermatológicas mais comuns encontradas na prática clínica. As opções de tratamento para esta dermatose são diversas, porém nem sempre são efetivas. A resistência microbiana frente aos antibióticos e os efeitos adversos das terapias convencionais têm feito pesquisadores investigarem tratamentos alternativos. Dessa forma, este trabalho teve por objetivo avaliar a ação da terapia fotodinâmica (TFD) mediada por curcumina, azul de metileno e clorina-e6 sobre biofilmes de bactérias provenientes de pacientes com acne vulgar. Foram realizadas de uma a quatro aplicações fotodinâmicas nos biofilmes e a identificação dos microrganismos presentes nas amostras foi realizada por PCR convencional, por amplificação de genes com primers espécie-específicos. A biocompatibilidade dos tratamentos foi avaliada em um modelo de co-cultura em três dimensões (3D) de pele. Os microrganismos identificados nas amostras clínicas foram Cutibacterium acnes, Staphylococcus epidermidis e Staphylococcus aureus. Os resultados mostraram que a TFD mediada por azul de metileno foi capaz de eliminar completamente as células dos biofilmes acneicos após três aplicações, enquanto que a TFD mediada por curcumina e clorina-e6, embora tenham apresentado reduções significativas, não eliminaram totalmente as células dos biofilmes, mesmo após quatro aplicações fotodinâmicas. Dessa forma, o sucesso das aplicações consecutivas pode ser dependente do fotossensibilizador. Os ensaios de biocompatibilidade mostraram que a TFD mediada pelos três fotossensibilizadores reduziram significativamente a viabilidade das células no modelo de cultivo 3D de pele e esse efeito pode ser dependente da fluência utilizada. Tomados em conjunto, os resultados obtidos nesse estudo revelaram que os três fotossensibilizadores estudados podem ser promissores para o desenvolvimento de protocolos clínicos eficientes no tratamento da acne vulgar.
Acne vulgaris is one of the most common dermatological diseases encountered in clinical practice. The treatment options for this dermatosis are diverse, but they are not always effective. Microbial resistance to antibiotics and the adverse effects of conventional therapies have led researchers to investigate alternative treatments. Thus, this study aimed to evaluate the action of photodynamic therapy (PDT) mediated by curcumin, methylene blue and chlorin-e6 on bacterial biofilms from patients with acne vulgaris. One to four photodynamic applications were performed on the biofilms and the identification of microorganisms present in the samples was performed by conventional PCR, by gene amplification with species-specific primers. The biocompatibility of the treatments was evaluated in a three-dimensional (3D) skin co-culture model. The microorganisms identified in the clinical samples were Cutibacterium acnes, Staphylococcus epidermidis and Staphylococcus aureus. The results showed that PDT mediated by methylene blue was able to completely eliminate cells from acne biofilms after three applications, while PDT mediated by curcumin and chlorin-e6, although they showed significant reductions, did not completely eliminate cells from biofilms even after four photodynamic applications. Thus, the success of consecutive applications may be dependent on the photosensitizer. Biocompatibility assays showed that PDT mediated by the three photosensitizers significantly reduced cell viability in the 3D skin culture model and this effect may be dependent on the fluence used. Taken together, the results obtained in this study revealed that the three photosensitizers studied may be promising for the development of efficient clinical protocols in the treatment of acne vulgaris.
Acne vulgaris is one of the most common dermatological diseases encountered in clinical practice. The treatment options for this dermatosis are diverse, but they are not always effective. Microbial resistance to antibiotics and the adverse effects of conventional therapies have led researchers to investigate alternative treatments. Thus, this study aimed to evaluate the action of photodynamic therapy (PDT) mediated by curcumin, methylene blue and chlorin-e6 on bacterial biofilms from patients with acne vulgaris. One to four photodynamic applications were performed on the biofilms and the identification of microorganisms present in the samples was performed by conventional PCR, by gene amplification with species-specific primers. The biocompatibility of the treatments was evaluated in a three-dimensional (3D) skin co-culture model. The microorganisms identified in the clinical samples were Cutibacterium acnes, Staphylococcus epidermidis and Staphylococcus aureus. The results showed that PDT mediated by methylene blue was able to completely eliminate cells from acne biofilms after three applications, while PDT mediated by curcumin and chlorin-e6, although they showed significant reductions, did not completely eliminate cells from biofilms even after four photodynamic applications. Thus, the success of consecutive applications may be dependent on the photosensitizer. Biocompatibility assays showed that PDT mediated by the three photosensitizers significantly reduced cell viability in the 3D skin culture model and this effect may be dependent on the fluence used. Taken together, the results obtained in this study revealed that the three photosensitizers studied may be promising for the development of efficient clinical protocols in the treatment of acne vulgaris.
Descrição
Palavras-chave
biofilmes polimicrobianos, terapia fotodinâmica, Cutibacterium acnes, acne vulgar, curcumina, azul de metileno, clorina-e6, polymicrobial biofilms, photodynamic therapy, acne vulgaris, curcumin, methylene blue, chlorin-e6