Ação antimicrobiana e citotoxicidade de extratos aquoso e glicólico de própolis sobre bactérias anaeróbias de importância odontológica
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Data
2018-11-27
Autores
Assis, Maria Angélica de Sá [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
As plantas medicinais e os fitoterápicos têm sido utilizados como coadjuvantes e alternativos no combate a diversas doenças com crescente frequência, no entanto, na Odontologia seu uso ainda é bastante limitado. Com isso, o objetivo deste estudo é avaliar a ação antimicrobiana dos extratos aquoso e glicólico de própolis verde sobre micro-organismos anaeróbios de interesse odontológico, bem como sua citotoxicidade, a fim de introduzir e incentivar o uso efetivo e sistemático desse fitoterápico em produtos como dentifrícios e enxaguatórios bucais no combate a cáries e doenças periodontais. Os extratos comerciais aquoso e glicólico de própolis foram obtidos das empresas Apis Flora e Mapric, respectivamente. Para avaliação da atividade antimicrobiana foram utilizadas cepas-padrão (ATCC) de Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis Porphyromonas gingivalis e Prevotella intermedia em cultura planctônica, verificando a concentração inibitória mínima e concentração microbicida mínima (CIM e CMM), segundo Clinical and Laboratory Standards Institute. Para biofilmes monotípicos, suspensões padronizadas (107 céls/mL) foram adicionadas em poços de microplacas e após 48 h em anaerobiose foram tratados com 3 concentrações do extrato de própolis (n=12) por 5 min. Foi incluído um controle positivo (solução fisiológica) e um controle negativo (clorexidina). O biofilme foi mensurado pelos testes MTT e Cristal Violeta. Para análise de citotoxicidade, queratinócitos humanos (HaCaT) foram cultivados e colocados em contato com os extratos por 5 min e 24h. Os dados foram analisados estatisticamente pelos testes ANOVA e Tukey Test (5%). Os resultados mostraram que os extratos tiveram ação antimicrobiana contra as suspensões planctônicas e os biofilmes monotípicos dos patógenos, sendo tão ou mais eficazes que a clorexidina. Quanto à citotoxicidade, observou-se diminuição da viabilidade celular dos queratinócitos humanos após a aplicação dos extratos, do mesmo modo que a clorexidina, sendo o extrato glicólico menos citotóxico que o aquoso. Com isto conclui-se que os extratos comerciais aquoso e glicólico de própolis verde tem ação antimicrobiana contra os micro-organismos anaeróbios orais estudados e apresentam potencial para serem utilizados nos tratamentos contra os referidos patógenos, já que no que se refere à citotoxicidade, se comportaram de forma semelhante à Clorexidina, cujo uso é conhecidamente seguro e eficaz.
Medicinal plants and herbal medicines have been used as adjuvants and alternatives in fighting various diseases with increasing frequency. However, in dentistry its use is still quite limited. Therefore, the objective of this study is to evaluate the antimicrobial action of the aqueous and glycolic extracts of green propolis on anaerobic microorganisms of dental interest, in order to introduce and encourage the effective and systematic use of this herbal medicine in products such as dentifrices and mouthwashes in the fight against tooth decay and periodontal diseases. Aqueous and glycolic commercial extracts of propolis were obtained from Apis Flora and Mapric companies, respectively.To evaluate the antimicrobial activity, standard strains (ATCC) of Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in planktonic culture was used, verifying the minimum inhibitory concentration and minimum microbicide concentration (MIC and CMM), according to Clinical and Laboratory Standards Institute. For monotypic biofilms, standardized suspensions (107 cells / mL) was added to microplate wells and after 48 h in anaerobiosis was treated with 3 concentrations of propolis extracts (n = 12) for 5 min. A positive control (saline solution) and a negative control (chlorhexidine) was included. The biofilm was measured by the MTT and Violet Crystal tests. For cytotoxicity analysis, human keratinocytes (HaCaT) were cultured and placed in contact with the extracts for 5 min and 24 h. Data were analyzed statistically by ANOVA and Tukey Test (5%). The results showed that the extracts had antimicrobial action against planktonic suspensions and monotypic pathogen biofilms, being as or more effective than chlorhexidine. As for cytotoxicity, the cellular viability of human keratinocytes was observed after application of the extracts, in the same way as chlorhexidine, the glycolic extract being less cytotoxic than aqueous. It is concluded that the aqueous and glycolic extracts of green propolis have an antimicrobial action against the studied oral anaerobic microorganisms and and have the potential to be used in the treatments against these pathogens, since in cytotoxicity they behaved in a way similar to chlorhexidine, the use of which is known to be safe and effective.
Medicinal plants and herbal medicines have been used as adjuvants and alternatives in fighting various diseases with increasing frequency. However, in dentistry its use is still quite limited. Therefore, the objective of this study is to evaluate the antimicrobial action of the aqueous and glycolic extracts of green propolis on anaerobic microorganisms of dental interest, in order to introduce and encourage the effective and systematic use of this herbal medicine in products such as dentifrices and mouthwashes in the fight against tooth decay and periodontal diseases. Aqueous and glycolic commercial extracts of propolis were obtained from Apis Flora and Mapric companies, respectively.To evaluate the antimicrobial activity, standard strains (ATCC) of Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in planktonic culture was used, verifying the minimum inhibitory concentration and minimum microbicide concentration (MIC and CMM), according to Clinical and Laboratory Standards Institute. For monotypic biofilms, standardized suspensions (107 cells / mL) was added to microplate wells and after 48 h in anaerobiosis was treated with 3 concentrations of propolis extracts (n = 12) for 5 min. A positive control (saline solution) and a negative control (chlorhexidine) was included. The biofilm was measured by the MTT and Violet Crystal tests. For cytotoxicity analysis, human keratinocytes (HaCaT) were cultured and placed in contact with the extracts for 5 min and 24 h. Data were analyzed statistically by ANOVA and Tukey Test (5%). The results showed that the extracts had antimicrobial action against planktonic suspensions and monotypic pathogen biofilms, being as or more effective than chlorhexidine. As for cytotoxicity, the cellular viability of human keratinocytes was observed after application of the extracts, in the same way as chlorhexidine, the glycolic extract being less cytotoxic than aqueous. It is concluded that the aqueous and glycolic extracts of green propolis have an antimicrobial action against the studied oral anaerobic microorganisms and and have the potential to be used in the treatments against these pathogens, since in cytotoxicity they behaved in a way similar to chlorhexidine, the use of which is known to be safe and effective.
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Palavras-chave
Extratos naturais, Própolis, Atividade antimicrobiana, Citotoxicidade, Bactérias Anaeróbias, Natural extracts, Propolis. Antimicrobian activity, Cytotoxicity, Anaerobic bacteria