Atividade antimicrobiana e antibiofilme de óleos essenciais contra micro-organismos orais
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Data
2018-11-22
Autores
Albuquerque, Yasmin Etienne
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Universidade Estadual Paulista (Unesp)
Resumo
Os objetivos deste trabalho foram: a) realizar uma revisão crítica sobre as plantas listadas na Relação Nacional de Plantas Medicinais de Interesse ao SUS (RENISUS) que apresentam evidência de atividade antimicrobiana contra micro-organismos cariogênicos (Publicação 1); b): avaliar e comparar a atividade antibacteriana e anti-aderente de óleos essenciais de plantas incluídas na RENISUS obtidos de duas fontes (Publicação 2); c) comparar o comportamento de biofilmes de Streptococcus mutans com biofilmes polimicrobianos e avaliar a citotoxicidade e a capacidade de óleos essenciais de plantas incluídas na RENISUS em interferir na maturação desses biofilmes (Publicação 3). Publicação 1: foram selecionados artigos que avaliaram extratos de plantas listadas na RENISUS contra micro-organismos cariogênicos. Apenas 17 das 71 plantas da RENISUS foram estudadas. Dos 24 estudos in vitro, apenas 6 realizaram testes com biofilmes. Apenas Lippia sidoides e Punica granatum foram avaliadas em estudos clínicos randomizados ou cruzados. Publicação 2: de cada espécie vegetal selecionada foram obtidas duas amostras de óleos essenciais de fontes diferentes (Fonte comercial: Empórios Laszlo ou Quinari; Fonte ex-situ: Banco de germoplasma). Foram utilizados micro-organismos orais de referência (Actinomyces naeslundii, Lactobacillus acidophilus, Streptococcus oralis, Streptococcus mutans, Streptococcus parasanguinis, Streptococcus salivarius e Streptococcus mitis). Para fins comparativos, foi utilizado o digluconato de clorexidina 0,12% (CHX). A concentração inibitória mínima (CIM) e bactericida mínima (CBM) dos óleos essenciais foram determinadas. Os óleos essenciais mais ativos (Cordia curassavica e Eugenia uniflora da Fonte comercial e Eugenia uniflora e Tagetes minuta da Fonte ex-situ) foram avaliados em concentrações sub-inibitórias quanto à sua atividade anti-aderente e caracterizados quimicamente. Os óleos essenciais de C. curassavica (Fonte comercial) e T. minuta (Fonte ex-situ) apresentaram atividade anti-aderente contra um maior número cepas. Os constituintes majoritários dos óleos essenciais são sesquiterpenos. Publicação 3: os óleos essenciais foram avaliados quanto à CIM e CBM de S. mutans e inóculo polimicrobiano proveniente de saliva humana. Os óleos essenciais mais ativos foram avaliados quanto à capacidade em interferir na maturação de biofilmes. Os biofilmes foram analisados quanto a sua acidogenicidade, composição microbiana e bioquímica. A citotoxicidade dos óleos essenciais ativos em biofilmes foi avaliada utilizando cultura de fibroblastos e queratinócitos. O óleo de E. uniflora de ambas as fontes foi o mais ativo e foi capaz de reduzir a viabilidade e a acidogenicidade dos biofilmes e apresentaram citotoxicidade menor ou igual a CHX. Biofilmes polimicrobianos apresentaram comportamento diferente em comparação à biofilmes de S. mutans (menor viabilidade, maior acidogenicidade e maior concentração de compostos bioquímicos). Conclusão geral: poucas plantas listadas na RENISUS foram avaliadas quanto ao seu potencial anticariogênico (Publicação 1). Os óleos essenciais de C. curassavica (Fonte comercial) e T. minuta (Fonte ex-situ) apresentaram melhor atividade anti-aderente. Óleos essenciais obtidos de fonte comercial podem ser tão efetivos quanto os óleos obtidos de coleções ex-situ (Publicação 2). O óleo essencial de E. uniflora de ambas as fontes foram efetivos em biofilmes. Biofilmes monotípicos podem superestimar a ação antimicrobiana de produtos naturais, sendo indicado então, para estudos in vitro, o uso de biofilmes polimicrobianos (Publicação 3).
The objectives of this work were: a) to carry out a critical review of the plants listed in the “Relação Nacional de Plantas Medicinais de Interesse ao SUS” (RENISUS) with evidence of antimicrobial activity against cariogenic microorganisms (Publication 1); b) to evaluate and compare antibacterial and anti-adherent activity of essential oils from plants included in RENISUS obtained from two sources (Publication 2); c) to compare the behavior of Streptococcus. mutans and polymicrobial biofilms and to evaluate the cytotoxicity and the ability of essential oils from plants included in RENISUS to interfere on the maturation of these biofilms (Publication 3). Publication 1: Original papers were searched that evaluated plants extracts listed in the RENISUS against cariogenic microorganisms. Only 17 plants of the 71 RENISUS plants were studied. Of the 24 in vitro studies, only 6 carried out tests with biofilms. Only Lippia sidoides and Punica granatum were evaluated in randomized or cross-over clinical trials. Publication 2: Two samples of essential oils from different sources were obtained from each selected plant species (Marketed source: Empório Laszlo or Quinari; Ex-situ source: Germoplasm bank). Reference microorganisms (Actinomyces naeslundii ATCC 19039, Lactobacillus acidophilus ATCC 4356, Streptococcus oralis ATCC 10557, Streptococcus mutans ATCC 35688, Streptococcus parasanguinis ATCC 903, Streptococcus salivarius ATCC 7073 and Streptococcus mitis ATCC 9811). For comparison purposes, chlorhexidine digluconate 0.12% (CHX) was used. The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of essential oils were determined. The most active essential oils (Cordia curassavica and E. uniflora from marketed sources and Eugenia uniflora and Tagetes minuta from ex-situ collection) were evaluated at sub-inhibitory concentrations for their anti-adherent activity and chemically characterized. The essential oils of C. curassavica (marketed source) and T. minuta (ex-situ source) showed anti-adherent activity against a larger number of reference microorganisms. The major constituents of essential oils are sesquiterpenes. Publication 3: essential oils were evaluated for CIM and CBM of S. mutans ATCC 35688 and polymicrobial inoculum from human saliva. The most active essential oils were evaluated for the ability to interfere in the maturation of biofilms. Biofilms were analyzed for their acidogenicity, microbiological and biochemical composition. The cytotoxicity of the most active essential oils was evaluated using fibroblasts and keratinocytes cultures. E. uniflora essential oil from both sources reduced biofilm viability and acidogenicity and showed cytotoxicity lower or equal to CHX. Polymicrobial biofilms and S. mutans biofilms showed different behavior (lower viability, higher acidogenicity and higher concentration of biochemical compounds). General conclusion: few plants listed in RENISUS were evaluated for their anticariogenic potential (Publication 1). The essential oils of C. curassavica (marketed source) and T. minuta (ex-situ source) showed the best anti-adherence activity. Essential oils obtained from marketed sources can be as effective as oils obtained from ex-situ collections (Publication 2). E. uniflora essential oils from both sources were effective in biofilms. Monotypic biofilms may overestimate the antimicrobial action of natural products then being indicated, in vitro studies, the use of polymicrobial biofilms (Publication 3).
The objectives of this work were: a) to carry out a critical review of the plants listed in the “Relação Nacional de Plantas Medicinais de Interesse ao SUS” (RENISUS) with evidence of antimicrobial activity against cariogenic microorganisms (Publication 1); b) to evaluate and compare antibacterial and anti-adherent activity of essential oils from plants included in RENISUS obtained from two sources (Publication 2); c) to compare the behavior of Streptococcus. mutans and polymicrobial biofilms and to evaluate the cytotoxicity and the ability of essential oils from plants included in RENISUS to interfere on the maturation of these biofilms (Publication 3). Publication 1: Original papers were searched that evaluated plants extracts listed in the RENISUS against cariogenic microorganisms. Only 17 plants of the 71 RENISUS plants were studied. Of the 24 in vitro studies, only 6 carried out tests with biofilms. Only Lippia sidoides and Punica granatum were evaluated in randomized or cross-over clinical trials. Publication 2: Two samples of essential oils from different sources were obtained from each selected plant species (Marketed source: Empório Laszlo or Quinari; Ex-situ source: Germoplasm bank). Reference microorganisms (Actinomyces naeslundii ATCC 19039, Lactobacillus acidophilus ATCC 4356, Streptococcus oralis ATCC 10557, Streptococcus mutans ATCC 35688, Streptococcus parasanguinis ATCC 903, Streptococcus salivarius ATCC 7073 and Streptococcus mitis ATCC 9811). For comparison purposes, chlorhexidine digluconate 0.12% (CHX) was used. The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of essential oils were determined. The most active essential oils (Cordia curassavica and E. uniflora from marketed sources and Eugenia uniflora and Tagetes minuta from ex-situ collection) were evaluated at sub-inhibitory concentrations for their anti-adherent activity and chemically characterized. The essential oils of C. curassavica (marketed source) and T. minuta (ex-situ source) showed anti-adherent activity against a larger number of reference microorganisms. The major constituents of essential oils are sesquiterpenes. Publication 3: essential oils were evaluated for CIM and CBM of S. mutans ATCC 35688 and polymicrobial inoculum from human saliva. The most active essential oils were evaluated for the ability to interfere in the maturation of biofilms. Biofilms were analyzed for their acidogenicity, microbiological and biochemical composition. The cytotoxicity of the most active essential oils was evaluated using fibroblasts and keratinocytes cultures. E. uniflora essential oil from both sources reduced biofilm viability and acidogenicity and showed cytotoxicity lower or equal to CHX. Polymicrobial biofilms and S. mutans biofilms showed different behavior (lower viability, higher acidogenicity and higher concentration of biochemical compounds). General conclusion: few plants listed in RENISUS were evaluated for their anticariogenic potential (Publication 1). The essential oils of C. curassavica (marketed source) and T. minuta (ex-situ source) showed the best anti-adherence activity. Essential oils obtained from marketed sources can be as effective as oils obtained from ex-situ collections (Publication 2). E. uniflora essential oils from both sources were effective in biofilms. Monotypic biofilms may overestimate the antimicrobial action of natural products then being indicated, in vitro studies, the use of polymicrobial biofilms (Publication 3).
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Palavras-chave
Plantas medicinais, Produtos com ação antimicrobiana, Placa dentária, Medicinal plants, Products with antimicrobial action, Dental plaque