Avaliação da eficácia de tt-farnesol liberado por novo sistema de liberação controlada de fármacos contra biofilmes orais patogênicos
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Data
2017-07-07
Autores
Rocha, Guilherme Roncari
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O desenvolvimento de terapias eficazes para o controle de biofilme oral é um desafio. Agentes tópicos usados para tratar ou prevenir doenças orais causadas por biofilme, geralmente, não são mantidos na boca por um período suficiente para exercer o seu potencial terapêutico. Portanto, estratégias que objetivem na retenção e liberação controlada de agentes ativos no local onde o biofilme virulento se desenvolve são necessárias. Assim, nanopartículas carreadoras (NPC) de fármaco que se aderem à hidroxiapatita, à película salivar e aos exopolissacarídeos da matriz extracelular de biofilme foram desenvolvidas. As NPC possuem elementos responsivos à mudanças de pH para a liberação controlada de agentes terapêuticos. Portanto, o objetivo deste estudo foi avaliar a eficácia de tratamento tópico utilizando sistema de liberação de fármacos carregado com tt-farnesol sobre película salivar e biofilme misto de Streptococcus mutans e Candida albicans em diferentes superfícies encontradas na cavidade bucal. Para tanto, foram formados biofilmes mistos sobre a superfície de zircônia (LAVA Frame, 3M), de resina acrílica (Lucitone 550, Dentsply) e de hidroxiapatita (controle). Foram aplicados 6 tratamentos tópicos com tempo de exposição de 5 minutos: PBS (tampão fosfato diluente de NPC); NPC-Livre; NPC + tt-farnesol; Veículo (PBS com 15% álcool etílico diluente de tt-farnesol isolado); e tt-farnesol. As análises realizadas foram: biomassa, exopolissacarídeos solúveis e insolúveis em água, pH do meio de cultura, população microbiana e análise da morfologia 3D via microscopia confocal. Os tratamentos com nanopartículas carreada associada ao farnesol mostraram redução na população de S. mutans e de exopolissacarídeos solúveis em água (os quais geram proteção para este biofilme), independente do substrato. Porém, esse tratamento não alterou a população do fungo e aumentou a quantidade de exopolissacarídeos insolúveis em água. Assim, a redução da população da bactéria e a menor proteção da mesma por exopolissacarídeos solúveis torna eficiente o tratamento associado à tecnologia de nanopartículas. Porém, é necessário adequar a estratégia para reduzir a população do fungo e de exopolissacarídeos insolúveis em água.
The development of efficacious therapies to control oral biofilms is challenging. Topical agents used to treat or prevent oral diseases are generally not retained in the mouth for sufficient duration to exert their full therapeutic potential. Thus, novel strategies for biofilm control focused on retention and sustained delivery of active agents at the site where virulent biofilm develops are warranted. Therefore, nanoparticle-carriers (NPC) that bind avidly to hydroxyapatite, to tooth-pellicle and exopolysaccharides were developed. NPC contains pH-responsive elements that facilitate controlled-release of therapeutic agents. Therefore, the aim of this study was to evaluate the efficacy of topical treatment using farnesol loaded delivery system against pathogenic mixed-species biofilm of Streptococcus mutans and Candida albicans on different surfaces found in oral cavity. Therefore, mixed-species biofilms were formed on different surfaces: zirconia (LAVA Frame, 3M), acrylic resin (Lucitone 550, Dentsply) and hydroxyapatite (control). Six topical treatments were applied on different surfaces for 5 minutes: PBS (phosphate buffer used to prepare NPC); NPC-free; NPC + tt-farnesol; Vehicle (phosphate buffer with 15% ethanol used to prepare tt-farnesol); and tt-farnesol. The assays performed were: biomass, watersoluble and -insoluble exopolysaccharides, pH of spent media, microbial population, and 3D architecture of biofilms by confocal microscopy. Nanoparticulate treatments associated with tt-farnesol showed a reduction in the amount of S. mutans population and amounts of water-soluble exopolysaccharides (which provides protection for biofilm), independent of the substrate. In addition, this treatment had no effect on C. albicans population and on amounts of water-insoluble exopolysaccharides. Therefore, the reduction of bacterial population and of protection of the microbial population by water-soluble exopolysaccharides demonstrated that the treatment associated with nanoparticle technology is efficient. However, it is necessary to improve the strategy to reduce also the fungal population and the water-insoluble exopolysaccharides
The development of efficacious therapies to control oral biofilms is challenging. Topical agents used to treat or prevent oral diseases are generally not retained in the mouth for sufficient duration to exert their full therapeutic potential. Thus, novel strategies for biofilm control focused on retention and sustained delivery of active agents at the site where virulent biofilm develops are warranted. Therefore, nanoparticle-carriers (NPC) that bind avidly to hydroxyapatite, to tooth-pellicle and exopolysaccharides were developed. NPC contains pH-responsive elements that facilitate controlled-release of therapeutic agents. Therefore, the aim of this study was to evaluate the efficacy of topical treatment using farnesol loaded delivery system against pathogenic mixed-species biofilm of Streptococcus mutans and Candida albicans on different surfaces found in oral cavity. Therefore, mixed-species biofilms were formed on different surfaces: zirconia (LAVA Frame, 3M), acrylic resin (Lucitone 550, Dentsply) and hydroxyapatite (control). Six topical treatments were applied on different surfaces for 5 minutes: PBS (phosphate buffer used to prepare NPC); NPC-free; NPC + tt-farnesol; Vehicle (phosphate buffer with 15% ethanol used to prepare tt-farnesol); and tt-farnesol. The assays performed were: biomass, watersoluble and -insoluble exopolysaccharides, pH of spent media, microbial population, and 3D architecture of biofilms by confocal microscopy. Nanoparticulate treatments associated with tt-farnesol showed a reduction in the amount of S. mutans population and amounts of water-soluble exopolysaccharides (which provides protection for biofilm), independent of the substrate. In addition, this treatment had no effect on C. albicans population and on amounts of water-insoluble exopolysaccharides. Therefore, the reduction of bacterial population and of protection of the microbial population by water-soluble exopolysaccharides demonstrated that the treatment associated with nanoparticle technology is efficient. However, it is necessary to improve the strategy to reduce also the fungal population and the water-insoluble exopolysaccharides
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Palavras-chave
Biofilmes, Streptococcus mutans, Candida albicans, Biofilms, Streptococcus mutans, Candida albicans