Análise da atividade antimicrobiana de probióticos e sua adesividade a bráquetes ortodônticos: estudo in vitro

Thayse Caroline de Abreu BRANDI; Amanda Nunes MONTEIRO; Hugo Leandro Azevedo da SILVA; Adriano Gomes da CRUZ; Lucianne Cople MAIA; Matheus Melo PITHON

doi:10.1590/1807-2577.09219

Original Article

Análise da atividade antimicrobiana de probióticos e sua adesividade a bráquetes ortodônticos: estudo in vitro

Analysis of probiotic antimicrobial activity and its adherence to orthodontic brackets: in vitro study

Thayse Caroline de Abreu BRANDI; Amanda Nunes MONTEIRO; Hugo Leandro Azevedo da SILVA; Adriano Gomes da CRUZ; Lucianne Cople MAIA; Matheus Melo PITHON

http://dx.doi.org/10.1590/1807-2577.09219 Rev. odontol. UNESP, vol.48, e20190092, 2019

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Resumo

Resumo: Introdução:

A presença de aparelho ortodôntico fixo dificulta a higienização e potencializa o acúmulo de biofilme bacteriano nas superfícies dentárias. O desenvolvimento de produtos que minimize isso é desejo de pesquisadores em todo o mundo.

Objetivo: Verificar a ação bacterapêutica de produtos lácteos contendo ou não probióticos sob pool de Streptococcus mutans (SM) (ATCC 25175) e S salivarius (SS) (ATCC 7073), além da adesão desses produtos à superfície de bráquetes ortodônticos.

Material e método: Pool de cepas ATCC de SM e SS foi formado e plaqueado sobre placa de Petri contendo meio de cultura brain heart infusion ágar (BHI). Após formação do meio, um orifício foi feito no centro da placa seguido do seu preenchimento com 150 µL dos produtos a serem testados, formando os seguintes grupos: GL - Leite bovino; GLP - Leite bovino com probiótico; GLF - Leite fermentado; e GLFP - Leite fermentado com probiótico. Na sequência, as placas foram incubadas por 48h, em estufa a 37ºC. A seguir, foi feita a medição do halo formado entre o produto e o meio com régua milimetrada. Já no disco de membrana, foi formado biofilme com o mesmo pool de cepas, sob discos de membrana. Em seguida, foi feita a diluição seriada contendo o produto de acordo com o grupo: P1 (água); P2 (L); P3 (LP); P4 (LFP), seguida do plaqueamento e a contagem total de micro-organismos. Para a adesividade dos produtos lácteos, bráquetes ortodônticos foram submergidos em cada solução (GL, GLP, GLF e GLFP) e foram incubadas a 37°C/24h. Posteriormente, cada bráquete foi transferido para um ependorf contendo solução salina estéril, que foi submetida a diluições seriadas, posteriormente incubadas a 37°C/48h sob microaerofilia para contagem das UFC/mL. Para análise dos dados, utilizaram-se os testes Levene, Shapiro-Wilk e Kruskal-Wallis. O nível de significância adotado foi de 5% (α = 0,05).

Resultado: Não houve formação de halo de inibição entre os produtos e o meio de cultura (p<0,05); no disco de membrana, não foram observadas diferenças estatísticas entre os grupos (p=0,679); os grupos tratados com leite bovino com probiótico e leite fermentado com probiótico apresentaram adesividade aos bráquetes ortodônticos (p=0,056).

Conclusão: Os achados do presente estudo permitem concluir que, em estudos in vitro, não foi possível verificar a bacterioterapia a partir de produtos lácteos contendo ou não probióticos em cepas de SM e SS.

Palavras-chave

Disco de membrana, halo de inibição, adesividade, bráquetes, probióticos

Abstract

Abstract: Introduction: The presence of a fixed orthodontic appliance makes cleaning difficult and enhances the accumulation of bacterial biofilm on dental surfaces. It is the desire of researchers worldwide.

Objective: To verify the bacteriotherapeutic action of dairy products containing or not probiotics under Streptococcus mutans (SM) (ATCC 25175) and S salivarius (SS) (ATCC 7073) pools, as well as their adhesion to the surface of orthodontic brackets.

Material and method: Pool of SM and SS ATCC strains was formed and plated on a petri dish containing brain heart infusion agar (BHI) culture medium. After formation of the medium a hole was made in the center of the plate followed by filling with 150 µL of the products to be tested, forming the following groups: GL - Bovine milk; GLP - Probiotic bovine milk; GLF - Fermented milk and GLFP - Probiotic fermented milk. Following the plates were incubated for 48h in an oven at 37°C. After that the halo formed between the product and the medium was measured with a millimeter ruler. Already in the membrane disc was formed biofilm with the same pool of strains, under membrane discs. After this the serial dilution containing the product was made according to the group: P1 (water); P2 (L); P3 (LP); P4 (LFP), followed by plating and total microorganism count. For the adhesiveness of dairy products, orthodontic brackets were submerged in each solution (GL, GLP, GLF and GLFP), which were incubated at 37°C/24h. Subsequently, each bracket was transferred to an ependorf containing sterile saline solution, which was subjected to serial dilutions, then incubated at 37°C/48h under microaerophilia for counting CFU/mL. For data analysis, the Levene, Shapiro-Wilk and Kruskal-Wallis tests were used. The adopted significance level was 5% (α=0.05).

Result: There was no inhibition halo formation between the products and the culture medium (p<0.05); In the membrane disc no statistical differences were observed between the groups (p=0.679); The group treated with probiotic bovine milk and probiotic fermented milk showed adherence to orthodontic brackets (p = 0.056).

Conclusion: The findings of the present study allow us to conclude that, in in vitro studies, it was not possible to verify bacteriotherapy from dairy products containing or not probiotics in SM and SS strains.

Keywords

Membrane disc, halo inhibition, adhesiveness, brackets, probiotics

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