Estudo da aplicação do extrato de própolis verde brasileiro em terapia fotodinâmica antimicrobiana contra Streptococcus mutan
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Data
2022-03-08
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Universidade Estadual Paulista (Unesp)
Resumo
O interesse pelos fotossensibilizadores naturais tem crescido nos últimos anos. Esse
crescimento está relacionado à capacidade de aderir ou atravessar a membrana
citoplasmática e ao potencial de produção de espécies reativas de oxigênio. Porém, até o
momento, não havia nenhum estudo que avaliasse o extrato de própolis como
fotossensibilizador contra micro-organismos. Dessa forma, o objetivo do presente
estudo foi avaliar in vitro a eficácia de dois extratos de própolis verde brasileira (EPVB)
como fotossensibilizadores para uso na terapia fotodinâmica antimicrobiana (TFDa)
contra Streptococcus mutans. A TFDa foi realizada em suspensões e biofilmes de S.
mutans (ATCC 35668) usando EPVB em concentrações subinibitórias. A eficiência do
TFDa foi avaliada por meio da avaliação da viabilidade celular. A produção de espécies
reativas de oxigênio foi avaliada usando sondas para detecção do radical hidroxila e do
oxigênio singleto. A citotoxicidade foi avaliada em queratinócitos orais por ensaio de
MTT. O estudo foi cego para o examinador que avaliou a viabilidade celular de S.
mutans e para o estatístico. A significância estatística foi considerada em α <0,05. Para
as suspensões, foi observada uma redução microbiana total de 7,86 log UFC / mL para
ambos os EPVBs. Para biofilmes, reduções superiores a 3 log UFC / mL foram
observadas para ambos os EPVBs. Os dois extratos produziram grande quantidade de
oxigênio singleto, sem diferença estatística entre eles (p> 0,05). Após a aplicação de
TFDa, apenas um EPVB apresentou citotoxicidade aceitável para queratinócitos
(73,07%), sem diferenças em comparação ao grupo controle vivo (p> 0,05). Os
resultados do presente estudo mostraram que os EPVBs têm potencial para serem
usados em TFDa contra S. mutans.
Interest in natural photosensitizers has grown in recent years. This growth is related to the ability to adhere to or cross the cytoplasmic membrane and the potential to produce reactive oxygen species. However, until now, there were no studies that evaluated the propolis extract as a photosensitizer against microorganisms. Thus, the aim of the present study was to evaluate in vitro the efficacy of two extracts of Brazilian green propolis (EPVB) as photosensitizers for use in antimicrobial photodynamic therapy (ADT) against Streptococcus mutans. TFDa was performed on suspensions and biofilms of S. mutans (ATCC 35668) using EPVB at subinhibitory concentrations. The efficiency of TFDa was evaluated through the evaluation of cell viability. The production of reactive oxygen species was evaluated using probes for the detection of hydroxyl radical and singlet oxygen. Cytotoxicity was evaluated in oral keratinocytes by MTT assay. The study was blinded to the examiner who evaluated S. mutans cell viability and to the statistician. Statistical significance was considered at α <0.05. For the suspensions, a total microbial reduction of 7.86 log CFU/mL was observed for both EPVBs. For biofilms, reductions greater than 3 log CFU/mL were observed for both EPVBs. The two extracts produced a large amount of singlet oxygen, with no statistical difference between them (p> 0.05). After the application of TFDa, only one EPVB showed acceptable cytotoxicity for keratinocytes (73.07%), with no differences compared to the live control group (p> 0.05). The results of the present study showed that EPVBs have the potential to be used in TFDa against S. mutans.
Interest in natural photosensitizers has grown in recent years. This growth is related to the ability to adhere to or cross the cytoplasmic membrane and the potential to produce reactive oxygen species. However, until now, there were no studies that evaluated the propolis extract as a photosensitizer against microorganisms. Thus, the aim of the present study was to evaluate in vitro the efficacy of two extracts of Brazilian green propolis (EPVB) as photosensitizers for use in antimicrobial photodynamic therapy (ADT) against Streptococcus mutans. TFDa was performed on suspensions and biofilms of S. mutans (ATCC 35668) using EPVB at subinhibitory concentrations. The efficiency of TFDa was evaluated through the evaluation of cell viability. The production of reactive oxygen species was evaluated using probes for the detection of hydroxyl radical and singlet oxygen. Cytotoxicity was evaluated in oral keratinocytes by MTT assay. The study was blinded to the examiner who evaluated S. mutans cell viability and to the statistician. Statistical significance was considered at α <0.05. For the suspensions, a total microbial reduction of 7.86 log CFU/mL was observed for both EPVBs. For biofilms, reductions greater than 3 log CFU/mL were observed for both EPVBs. The two extracts produced a large amount of singlet oxygen, with no statistical difference between them (p> 0.05). After the application of TFDa, only one EPVB showed acceptable cytotoxicity for keratinocytes (73.07%), with no differences compared to the live control group (p> 0.05). The results of the present study showed that EPVBs have the potential to be used in TFDa against S. mutans.
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Palavras-chave
Streptococcus mutans, Produtos com ação antimicrobiana, Fármacos fotossensibilizantes, Própole, Streptococcus mutans, Products with antimicrobial action, Photosensitizing agents, Propolis