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Master's Dissertation
DOI
https://doi.org/10.11606/D.25.2013.tde-14082013-094509
Document
Author
Full name
Bruno Lara Zarella
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 2013
Supervisor
Committee
Buzalaf, Marilia Afonso Rabelo (President)
Prakki, Anuradha
Wang, Linda
Title in Portuguese
Efeito de resinas experimentais contendo inibidores de proteases da matriz sobre gelatinases e colagenases
Keywords in Portuguese
Catequina
Clorexidina
Inibidores de MMP
Liberação de drogas
Abstract in Portuguese
A evolução das resinas compostas fez com que esses materiais passassem a ter uma durabilidade maior e características estéticas muito boas, mas o risco de cárie recorrente é ainda um problema a ser resolvido. Na tentativa de solucionar esse problema, estudos vêm sendo conduzidos na tentativa de se formularem resinas compostas contendo agentes antibacterianos, como é o caso da incorporação de clorexidina (CHX). Outro fato que impede a longevidade deste material é a degradação de matriz de colágeno por proteases ativadas por pH ácido. Para tentar contornar esse problema, a adição de clorexidina, assim como Epigallocatechin gallate (EGCg), clássicos antibacterianos e inibidores de proteases da matriz , como as metaloproteinases da matriz (MMP) a resinas, poderia melhorar a eficácia destes materiais como substitutos de dentina em procedimentos restauradores, aumentando a longevidade do tratamento restaurador, mediante preservação das propriedades mecânicas do material. Assim, o objetivo desse estudo é avaliar o poder de inibição de resinas experimentais contendo inibidores conhecidos de proteases da matriz sobre gelatinases e colagenase. Para isso, copolímeros experimentais foram preparados combinando Bis-GMA com o diluente TEGDMA (70/30 mol%). Com exceção do copolímero placebo (sem drogas), EGCg ou CHX foram incorporados a 1% em peso isoladamente ou em combinação, a 0,5% em peso cada. Amostras contendo EGCg, CHX ou EGCg e CHX concentradas 10X foram obtidas do armazenamento de espécimes polimerizados da resina experimental em água deionizada (1 mL) após o período de 24h a 37°C e sua posterior concentração. O efeito da ação dos inibidores foi checado por zimografia e confirmado por um ensaio enzimático específico para colagenases e gelatinases. Os dados passaram por teste de homogeneidade (Bartlett) e normalidade (Kolmogorov-Smirnov) e foram avaliados por ANOVA a 2 critérios, seguido pelo teste de Bonferroni para comparações individuais (p<0,05). Os resultados do presente estudo, mostraram que, in vitro, a liberação de EGCg e CHX incorporados em resinas é capaz de reduzir a atividade gelatinolítica das MMPs -2 e -9, bem como a atividade da colagenase bacteriana, sugerindo um efeito potencial no aumento da longevidade de restaurações de resinas. Com isso, podemos afirmar que a liberação de ativos de resinas experimentais é possível e que esses ativos são capazes de inibir as MMPs, assim sugerindo um novo substituto para dentina em procedimentos restauradores.
Title in English
Effect of experimental resins containing protease inhibitors on gelatinases and collagenase
Keywords in English
Catechin
Chlorhexidine
Drug release
MMP inhibitors
Abstract in English
The evolution of composite resins made these materials to have a greater durability and very good esthetics characteristics, but the risk of recurrent caries is still a problem to be solved. In the attempt to solve this problem, studies are being conducted with the purpose to formulate composite resins containing antibacterial agents, such as chlorhexidine (CHX). Another fact that prevents the longevity of this material is the degradation of the collagen matrix by the proteases activated by acidic pH. In order to solve this problem, the addition of chlorhexidine and/or Epigallocatechin gallate (EGCg), classical antibacterial agents and inhibitors of matrix proteases, such as matrix metalloproteinases (MMP) in resins, could improve the efficacy of these materials as dentin substitutes in restorative procedures, increasing the longevity of the restorative treatment, while preserving the mechanical properties of the material. Thus, the aim of this study is to evaluate the ability of experimental resins containing known matrix protease inhibitors on the inhibition of gelatinases and collagenase. For this purpose, experimental copolymers were prepared combining Bis-GMA with the diluent TEGDMA (70/30 mol%). Except for the placebo copolymer (drug free), EGCg or CHX were incorporated at 1% in weight, isolated or in combination (0.5% in weight each). Samples containing EGCg, CHX or EGCg and CHX concentrated 10X were obtained after storage of polymerized specimens of the experimental resin in deionized water (1 mL) after the period of 24 h, at 37°C and after that were concentra. The effect of the action of the inhibitors was checked by zymography and confirmed by an enzymatic test specific for collagenases and gelatinases. The data passed in the tests of homogeneity (Bartlett test) and normality (Kolmogorov-Smirnov test), and were evaluated by 2-way ANOVA, followed by Bonferroni test for individual comparisons (p<0.05). The results of this study showed that the in vitro release of EGCG and CHX incorporated in resins was able to reduce the gelatinolytic activity of MMPs-2 and -9 and bacterial collagenase activity, suggesting a potential effect in increasing the longevity of resin restorations. It can be concluded that the release of drugs from experimental resins is possible and that these drugs are able to inhibit MMPs, thereby suggesting a new substitute for dentin in restorative procedures.
 
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Publishing Date
2013-08-15
 
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