Potencial antimicrobiano e biocompatibilidade de óleos essenciais em associação a biomateriais à base de silicato tricálcico
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Data
2020-03-23
Autores
Castro Núñez, Gabriela Mariana
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Os cimentos reparadores endodônticos à base de silicato de cálcio apresentam biocompatibilidade e bioatividade. Porém, sua ação antimicrobiana é limitada. O acréscimo de óleos essencias pode favorecer as propriedades antimicrobianas de materiais. Os óleos essenciais são substâncias de origem vegetal com propriedades antimicrobiana, antiviral, antifúngica, antitoxigênica, antiparasitária e inseticida. Este estudo teve como objetivo avaliar a associação de diferentes óleos essenciais nas propriedades físico-químicas, citocompatibilida, potencial bioativo e atividade antibiofilme dos cimentos reparadores à base de silicato de cálcio, MTA Repair HP (MTA HP) e Biodentine (BIO). Publicação 1: avaliou a associação de 1,5% de Farnesol (FAR). Publicação 2: analisou o acréscimo de 1% de Thyme Essential Oil (TEO). Publicação 3: associou 1% de lemongrass oil (LEO). As propriedades físico-químicas de tempo de presa, solubilidade após 7 dias de imersão em água destilada e pH em 1, 3, 7, 14 e 21 dias foram avaliadas. A citocompatibilidade foi avaliada por meio dos testes de methyltetrazolium (MTT) e Vermelho Neutro (VN) após exposição das células Saos-2 às diferentes diluições dos eluídos dos materiais por 24h. A bioatividade foi avaliada pela atividade enzimática da fosfatase alcalina (ALP) nos períodos de 1, 3 e 7 dias. Nódulos de mineralização foram também quantificados após 21 dias pela coloração com vermelho de Alizarina (ARS). A capacidade de inibição de Metaloproteinase de Matriz (MMP) 2 e 9 foi mensurada por fluorescência. A capacidade antibiofilme foi avaliada pelo teste de contato direto modificado (TCDM) sobre biofilme mono- e dual-espécies de Enterococcus faecalis e/ou Candida albicans, formados em dentina bovina. Os dados foram submetidos aos testes estatísticos Shapiro-Wilk, ANOVA e Tukey ou Bonferroni, com nível de significância de 5%. Todos os materiais tiveram tempo de presa semelhante e mostraram capacidade de alcalinização do meio em todos os períodos avaliados. BIO mostrou perda de massa puro ou associado ao FAR, TEO e LEO, enquanto que MTAHP ganhou massa com ou sem as associações. Todos os materiais mostraram citocompatibilidade e aumento na atividade de ALP nos períodos avaliados. Os cimentos avaliados foram capazes de induzir a produção de nódulos de mineralização. Os materiais não mostraram atividade anti-MMP-9, no entanto, mostraram inibição de MMP-2. A adição de FAR, TEO e LEO aumentou a capacidade antibiofilme mono e dual-espécie sobre E. faecalis e/ou C. albicans dos materiais. Conclui-se que a adição de óleos essenciais aos cimentos MTAHP e BIO melhora a atividade antibiofilme, sem alterar significativamente suas propriedades físico-químicas, mantendo a citocompatibilidade e potencial bioativo dos cimentos, e mostrando potencial para maiores estudos para uma futura aplicação clínica.
Calcium silicate-based endodontic repair cements have biocompatibility and bioactivity. However, its antimicrobial action is limited. The addition of essential oils can favor the antimicrobial properties of materials. Essential oils are substances of plant origin with antimicrobial, antiviral, antifungal, antitoxigenic, antiparasitic and insecticide properties. This study aimed to evaluate the association of different essential oils in the physico-chemical properties, cytocompatibility, bioactive potential and antibiofilm activity of calcium silicatebased cements, MTA Repair HP (MTAHP) and Biodentine (BIO). publication 1: evaluated the association of 1.5% of Farnesol (FAR). Publication 2: analyzed the increase of 1% of Thyme Essential Oil (TEO). Publication 3: associated 1% lemongrass oil (LEO). The physicochemical properties of setting time, solubility after 7 days of immersion in distilled water and pH at 1, 3, 7, 14 and 21 days were evaluated. Cytocompatibility was assessed using methyltetrazolium (MTT) and Neutral Red (NR) tests after exposure of Saos-2 cells to different dilutions of the extracts of the materials for 24h. Bioactivity was assessed by the enzymatic activity of alkaline phosphatase (ALP) in the periods of 1, 3 and 7 days. Mineralization nodules were also quantified after 21 days by staining with Alizarin red (ARS). The inhibition capacity of Matrix Metalloproteinase (MMP) 2 and 9 was measured by fluorescence. The antibiofilm activity was assessed by the modified direct contact test (MDCT) on biofilm mono- and dual-species of Enterococcus faecalis and/or Candida albicans, formed in bovine dentin. The data were submitted to Shapiro-Wilk, ANOVA and Tukey or Bonferroni statistical tests, with a significance level of 5%. All evaluated materials had a similar setting time and showed the ability to alkalize the medium in all periods. BIO showed mass loss when pure or associated with FAR, TEO and LEO, while MTAHP gained mass with or without the associations. All materials showed cytocompatibility and increased ALP activity in the evaluated periods. The evaluated cements were able to induce the production of mineralized nodules. The materials did not show anti-MMP-9 activity, however, they showed inhibition of MMP-2. The addition of FAR, TEO and LEO increased the mono and dual-species antibiofilm activity against E. faecalis and / or C. albicans of the materials. It is concluded that the addition of essential oils to repair cements, MTAHP and BIO, improves the antibiofilm activity, without significantly altering its physical-chemical properties, maintaining the cytocompatibility and bioactive potential of the cements, and showing potential for clinical application.
Calcium silicate-based endodontic repair cements have biocompatibility and bioactivity. However, its antimicrobial action is limited. The addition of essential oils can favor the antimicrobial properties of materials. Essential oils are substances of plant origin with antimicrobial, antiviral, antifungal, antitoxigenic, antiparasitic and insecticide properties. This study aimed to evaluate the association of different essential oils in the physico-chemical properties, cytocompatibility, bioactive potential and antibiofilm activity of calcium silicatebased cements, MTA Repair HP (MTAHP) and Biodentine (BIO). publication 1: evaluated the association of 1.5% of Farnesol (FAR). Publication 2: analyzed the increase of 1% of Thyme Essential Oil (TEO). Publication 3: associated 1% lemongrass oil (LEO). The physicochemical properties of setting time, solubility after 7 days of immersion in distilled water and pH at 1, 3, 7, 14 and 21 days were evaluated. Cytocompatibility was assessed using methyltetrazolium (MTT) and Neutral Red (NR) tests after exposure of Saos-2 cells to different dilutions of the extracts of the materials for 24h. Bioactivity was assessed by the enzymatic activity of alkaline phosphatase (ALP) in the periods of 1, 3 and 7 days. Mineralization nodules were also quantified after 21 days by staining with Alizarin red (ARS). The inhibition capacity of Matrix Metalloproteinase (MMP) 2 and 9 was measured by fluorescence. The antibiofilm activity was assessed by the modified direct contact test (MDCT) on biofilm mono- and dual-species of Enterococcus faecalis and/or Candida albicans, formed in bovine dentin. The data were submitted to Shapiro-Wilk, ANOVA and Tukey or Bonferroni statistical tests, with a significance level of 5%. All evaluated materials had a similar setting time and showed the ability to alkalize the medium in all periods. BIO showed mass loss when pure or associated with FAR, TEO and LEO, while MTAHP gained mass with or without the associations. All materials showed cytocompatibility and increased ALP activity in the evaluated periods. The evaluated cements were able to induce the production of mineralized nodules. The materials did not show anti-MMP-9 activity, however, they showed inhibition of MMP-2. The addition of FAR, TEO and LEO increased the mono and dual-species antibiofilm activity against E. faecalis and / or C. albicans of the materials. It is concluded that the addition of essential oils to repair cements, MTAHP and BIO, improves the antibiofilm activity, without significantly altering its physical-chemical properties, maintaining the cytocompatibility and bioactive potential of the cements, and showing potential for clinical application.
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Palavras-chave
Materiais Biocompatíveis, Propriedades Físicas e Químicas, Óleos Voláteis, Biofilmes, Cândida albicans, Enterococcus faecalis, Endodontia, Biocompatible Materials, Physical and Chemical Properties, Biofilms, Candida albicans, Enterococcus faecalis, Endodontics, Oils, Volatile