Extrato da casca de Punica granatum potencializa o efeito anti cárie de um enxaguatório bucal contendo trimetafosfato de sódio e flúor
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Data
2020-03-11
Autores
Fernandes, Gabriela Lopes [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A cárie dentária está entre as principais e mais comuns doenças bucais. É causada por ácidos produzidos pelo biofilme microbiano que levam à desmineralização do esmalte. A prevenção e controle dessa doença crônica consistem na desorganização periódica do biofilme e na promoção da remineralização dentária. Para resolver esse problema, associamos o extrato de casca de Punica granatum (romã) (PPE) ao trimetafosfato de sódio (TMP) e fluoreto (F) em formulações para uso como enxaguatório bucal, e avaliamos sua eficácia na redução do processo de desmineralização do esmalte dental, bem como seu potencial anti biofilme contra importantes patógenos orais presentes na cárie dentária (Streptococcus mutans ATCC 25175 e Candida albicans ATCC 10231). Blocos de esmalte bovino (4 mm × 4 mm) selecionados por dureza superficial inicial (SHi) foram alocados aleatoriamente de acordo com grupos de tratamentos de formulação (n = 12 / grupo): ETF1 (3,0% PPE + 0,2% TMP + 100ppmF), TF1 (0,2% TMP + 100ppmF), ETF2 (3,0% PPE + 0,3% TMP + 225ppmF), TF2 (0,3% TMP + 225ppmF), F1 (100 ppmF), F2 (225 ppmF) e P (formulação sem E / T / F - placebo). Os blocos foram tratados 2x / dia com cada formulação e submetidos a cinco ciclos de pH (soluções desmineralizantes / remineralizantes) a 37° C. A seguir, determinaram-se a dureza superficial final (SHf), a dureza integrada da subsuperfície de perda (ΔKHN) padronizada e as concentrações de fluoreto de esmalte (F) de cálcio (Ca) e fósforo (P). A porcentagem de perda de dureza superficial (% SH) foi calculada (% SH = [(SHf - SHi) / SHi)] x 100), e as formulações que promoveram menores porcentagens de desmineralização do esmalte (% SH) e suas contrapartes foram selecionadas para os ensaios anti biofilme, bem como a formulação contendo apenas PPE (E). Para isso, ETF2 e TF2 (% SH = -34,5% e -53,1%, respectivamente), e a formulação E foram usadas para tratar por 1 ou 10 minutos biofilmes duplos de C. albicans e S. mutans crescidos por 24 horas em discos de hidroxiapatita (HA). A desmineralização da superfície do esmalte foi menor nas amostras tratadas com a formulação ETF2, resultando em uma diminuição de 46% na% SH em comparação com a F2. Novamente, a capacidade de reduzir o corpo da lesão (ΔKHN) foi maior (~ 26%) com ETF2 em relação a F2, e F2 proporcionou a maior concentração de F, Ca e P na superfície do esmalte. Entre as formulações de enxaguatório bucal ETF2, TF2 e E, as maiores taxas de redução de células viáveis foram exibidas tratando o biofilme com ETF2 por 10 minutos, independentemente do microrganismo testado. Em conclusão, a adição de PPE (3%) em enxaguatórios bucais contendo TMP (0,3) e F (225ppm) promoveu uma diminuição considerável no mineral sem perda de esmalte dental, além de reduzir consideravelmente o biofilme cariogênico formado por S. mutans e C albicans. Assim, cria uma perspectiva promissora para o desenvolvimento de um produto comercial dental sem álcool com os benefícios de milênios reconhecidos à saúde do Punica granatum.
Dental caries is among the main and most common oral diseases. It is caused by acids produced by microbial biofilm that lead to enamel demineralization. The prevention and control of this chronic disease consist of periodic disorganization of the biofilm and the promotion of dental remineralization. To address this problem, we associate Punica granatum (pomegranate) peel extract (PPE) with sodium trimemtaphosphate (TMP), and fluoride (F) in formulations for being used as mouthwash, and evaluate its efficacy on reducing dental enamel demineralization process as well as its antibiofilm potential against important oral pathogens present in dental caries (Streptococcus mutans ATCC 25175 and Candida albicans ATCC 10231). Bovine enamel blocks (4 mm × 4 mm) selected by initial surface hardness (SHi) were randomly allocated according to groups of formulation treatments (n= 12/group): ETF1 (3.0%PPE+0.2%TMP+100ppmF), TF1 (0.2%TMP+100ppmF), ETF2 (3.0%PPE+0.3%TMP+225ppmF), TF2 (0.3%TMP+225ppmF), F1 (100 ppmF), F2 (225 ppmF), and P (formulation without E/T/F - placebo). The blocks were treated 2×/day with each formulation and submitted to five pH cycles (demineralizing/remineralizing solutions) at 37°C. Next, final surface hardness (SHf), integrated loss subsurface hardness (ΔKHN), and enamel fluoride (F) calcium (Ca) and phosphorus (P) concentrations were determined. The percentage of surface hardness loss (%SH) was calculated (%SH = [(SHf - SHi)/SHi)] x 100), and the formulations which promoted lower percentages of enamel demineralization (%SH) and its counterparts were selected to the antibiofim assays, as well as formulation containing only PPE (E). For that, ETF2 and TF2 (%SH= -34.5% and -53.1% respectively), and formulation E were used to treat for 1 or 10 minutes dual biofilms of C. albicans and S. mutans grown for 24 hours on hydroxyapatite discs (HA). Demineralization of the enamel surface was lower in samples treated with formulation ETF2, resulting in a 46% decrease in %SH in comparison with F2. Again, the capacity to reduce the lesion body (ΔKHN) was higher (~ 26%) with ETF2 in relation to F2, and F2 provided the highest concentration of F, Ca and P in enamel surface. Amongst the mouthwash formulations ETF2, TF2 and E, the highest rates of viable cells reduction were exhibited by treating biofilm with ETF2 for 10 minutes regardless of the microorganism tested. In conclusion, the addition of PPE (3%) in mouthwashes containing TMP (0.3) and F (225ppm) promoted a considerably decrease in the mineral loss of dental enamel besides considerable reducing cariogenic biofilm formed by S. mutans and C. albicans. It thus creates a promising prospect for the development of an alcohol free dental commercial product with the millennial recognized health benefits of Punica granatum.
Dental caries is among the main and most common oral diseases. It is caused by acids produced by microbial biofilm that lead to enamel demineralization. The prevention and control of this chronic disease consist of periodic disorganization of the biofilm and the promotion of dental remineralization. To address this problem, we associate Punica granatum (pomegranate) peel extract (PPE) with sodium trimemtaphosphate (TMP), and fluoride (F) in formulations for being used as mouthwash, and evaluate its efficacy on reducing dental enamel demineralization process as well as its antibiofilm potential against important oral pathogens present in dental caries (Streptococcus mutans ATCC 25175 and Candida albicans ATCC 10231). Bovine enamel blocks (4 mm × 4 mm) selected by initial surface hardness (SHi) were randomly allocated according to groups of formulation treatments (n= 12/group): ETF1 (3.0%PPE+0.2%TMP+100ppmF), TF1 (0.2%TMP+100ppmF), ETF2 (3.0%PPE+0.3%TMP+225ppmF), TF2 (0.3%TMP+225ppmF), F1 (100 ppmF), F2 (225 ppmF), and P (formulation without E/T/F - placebo). The blocks were treated 2×/day with each formulation and submitted to five pH cycles (demineralizing/remineralizing solutions) at 37°C. Next, final surface hardness (SHf), integrated loss subsurface hardness (ΔKHN), and enamel fluoride (F) calcium (Ca) and phosphorus (P) concentrations were determined. The percentage of surface hardness loss (%SH) was calculated (%SH = [(SHf - SHi)/SHi)] x 100), and the formulations which promoted lower percentages of enamel demineralization (%SH) and its counterparts were selected to the antibiofim assays, as well as formulation containing only PPE (E). For that, ETF2 and TF2 (%SH= -34.5% and -53.1% respectively), and formulation E were used to treat for 1 or 10 minutes dual biofilms of C. albicans and S. mutans grown for 24 hours on hydroxyapatite discs (HA). Demineralization of the enamel surface was lower in samples treated with formulation ETF2, resulting in a 46% decrease in %SH in comparison with F2. Again, the capacity to reduce the lesion body (ΔKHN) was higher (~ 26%) with ETF2 in relation to F2, and F2 provided the highest concentration of F, Ca and P in enamel surface. Amongst the mouthwash formulations ETF2, TF2 and E, the highest rates of viable cells reduction were exhibited by treating biofilm with ETF2 for 10 minutes regardless of the microorganism tested. In conclusion, the addition of PPE (3%) in mouthwashes containing TMP (0.3) and F (225ppm) promoted a considerably decrease in the mineral loss of dental enamel besides considerable reducing cariogenic biofilm formed by S. mutans and C. albicans. It thus creates a promising prospect for the development of an alcohol free dental commercial product with the millennial recognized health benefits of Punica granatum.
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Palavras-chave
Punica granatum, Polifosfatos, Desmineralização, Esmalte dental, Antimicrobiano, Polyphosphates