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Master's Dissertation
DOI
https://doi.org/10.11606/D.25.2021.tde-06122021-130157
Document
Author
Full name
Caroline Gomes Carvalho
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 2021
Supervisor
Committee
Santos, Paulo Sergio da Silva (President)
Costa, Cyrene Piazera Silva
Magalhães, Ana Carolina
Medeiros, Frederico Buhatem
Title in Portuguese
Avaliação do efeito antimicrobiano da biguanida de polihexametileno (PHMB) comparada com digluconato de clorexidina (CHX) sobre um biofilme de microcosmos de saliva: estudo in vitro
Keywords in Portuguese
Antissépticos bucais
Biguanida de polihexametileno
Clorexidina
Abstract in Portuguese
A biguanida de polihexametileno (PHMB) é quimicamente semelhante ao digluconato de clorexidina (CHX), ambas biguanidas, porém cadeias terminais distintas, o que proporciona diferença na atividade terapêutica e perfil toxicológico. O objetivo desta pesquisa foi avaliar e comparar o efeito antimicrobiano da solução de PHMB com a CHX sobre a viabilidade de biofilmes de microcosmos de saliva. Para a formação do inóculo, foi coletada a saliva de 4 indivíduos, e selecionadas as 3 amostras mais heterogêneas para o pool bacteriano. A saliva artificial de McBain modificada com sacarose foi utilizada como meio de crescimento. Amostras de esmalte bovino (n=180) foram preparadas para a formação do biofilme com 5 dias de crescimento, para posteriormente serem tratadas com as soluções em concentrações comerciais (PHMB 0.2% e CHX 0.12%) e não comerciais (PHMB 0.1% e CHX 0.06%). Após os tratamentos, realizou-se avaliação imediata e tardia (24h após o tratamento), sobre as Unidades Formadoras de Colônias (UFC), as concentrações de L-lactato, análise da resazurina, e a vitalidade de bactérias vivas, matriz extracelular e polissacarídeos extracelulares em microscopia confocal de varredura a laser. Os testes não-paramétricos de Kruskal-Wallis e post-hoc de Dunn foram realizados sobre os grupos (p<0,05). As concentrações não comerciais (CHX 0.06 e PHMB 0.1) apresentaram maior potencial antimicrobiano no momento imediato sobre as cepas de microrganismos estudados, sendo que a CHX 0.06 se sobressaiu em relação a PHMB 0.1. Para a substantividade, a concentração comercial da CHX (0.12%) apresentou maior eficácia, fato que pode justificar seu uso corriqueiro na prática clínica. Enquanto a concentração não comercial da PHMB (0.1%) apresentou maior substantividade em relação àquela disponibilizada comercialmente (PHMB 0.2). As soluções de PHMB apresentaram efeito antimicrobiano promissores no momento imediato contra os microrganismos cariogênicos, (Estreptococos mutans e Lactobacilos totais); e sobre a estrutura do biofilme de microcosmos de saliva, devido a degradação da matriz extracelular e redução dos polissacarídeos extracelulares. No entanto, o enxaguatório de CHX se sobressaiu quanto ao efeito da substantividade sobre a ecologia de biofilmes de microcosmos salivares.
Title in English
Evaluation of the antimicrobial effect of polyhexamethylenebiguanide (PHMB) compared with chlorhexidinedigluconate (CHX) on a saliva microcosm biofilm: in vitro study
Keywords in English
Chlorhexidine
Mouthwashes
Polyhexamethylen-biguanide
Abstract in English
Polyhexamethylene biguanide (PHMB) is chemically similar to chlorhexidine gluconate (CHX), both biguanides, but with distinct terminal chains, which provide a difference in therapeutic activity and toxicological profile. This research aimed to evaluate and compare the antimicrobial effect of PHMB solution with CHX on the viability of saliva microcosm biofilms. For inoculum formation, saliva from 4 individuals was collected, and the 3 most heterogeneous samples were selected for the bacterial pool. McBain's artificial saliva modified with sucrose was used as the growth. Bovine enamel samples (n=180) were prepared for biofilm formation after 5 days of growth, to be later treated with solutions in commercial (PHMB 0.2% and CHX 0.12%) and non-commercial (PHMB 0.1% and CHX 0.06%) concentrations. After the treatments, an immediate and late evaluation (24h after treatment) was carried out on the Colony Forming Units (CFU), L-lactate concentrations, resazurin assay, and the vitality of live bacteria, extracellular matrix, and polysaccharides extracellular cells in confocal laser scanning microscopy. The non-parametric Kruskal-Wallis and Dunn posthoc tests were performed on the groups (p<0.05). The non-commercial concentrations (CHX 0.06 and PHMB 0.1) showed greater antimicrobial potential in the immediate moment on the strains of microorganisms studied, and CHX 0.06 stood out concerning PHMB 0.1. For substantivity, the commercial concentration of CHX (0.12%) was more effective, a fact that may justify its common use in clinical practice. While the non-commercial concentration of PHMB (0.1%) was more substantial than commercially available (PHMB 0.2). The PHMB solutions showed promising antimicrobial effect immediately against cariogenic microorganisms (Streptococci mutans and total Lactobacilli); and on the biofilm structure of saliva microcosms, due to extracellular matrix degradation and reduction of extracellular polysaccharides. However, the CHX mouthwash excelled in terms of the effect of substantivity on the ecology of salivary microcosm biofilms.
 
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Publishing Date
2021-12-06
 
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