OBJECTIVE: This study evaluated the inhibitory effect of proanthocyanidin (PA), a natural collagen cross-linker, on soluble and matrix-bound proteases, which are responsible for progressive degradation of exposed collagen fibrils within the hybrid layer and resin-dentine bond failure over time. METHODS: The inhibitory effects of PA (1%, 2%, 3%, 4.5% and 6%) on soluble recombinant matrix metalloproteinases (MMP-2, -8 and -9) and cysteine cathepsins (cathepsin B and K) were evaluated using MMP and cysteine cathepsins fluorometric assay kits. Chlorhexidine (CHX) was used as an inhibitor control. The effect of PA on endogenous matrix-bound proteases was examined by determining the change in dry mass of demineralized dentine beams and solubilized collagen peptides over 30 days. Two-way ANOVA and Tukey multiple comparison tests were used to analyze the effect of PA and proteases on the percentage inhibition of soluble proteases (α=0.05). Kruskal-Wallis one-way ANOVA and Dunn's multiple comparison tests were used to analyse the effect of PA on loss of dry mass and hydroxyproline content over time (α=0.05). RESULTS: Proanthocyanidin inactivated more than 90% of soluble recombinant MMP-2, -8 and -9 and around 75-90% of cysteine cathepsin B and K, which was significantly higher than CHX (P<0.05). The inhibition of endogenous proteases by PA increased in a dose-dependent manner. The loss of dry mass and hydroxyproline release in the medium over time was the lowest in dentine beams pretreated with PA<CHX<control (P<0.05). CONCLUSION: Proanthocyanidin exhibited both dentine MMP and cysteine cathepsins inhibition, which was higher than chlorhexidine.
OBJECTIVE: This study evaluated the inhibitory effect of proanthocyanidin (PA), a natural collagen cross-linker, on soluble and matrix-bound proteases, which are responsible for progressive degradation of exposed collagen fibrils within the hybrid layer and resin-dentine bond failure over time. METHODS: The inhibitory effects of PA (1%, 2%, 3%, 4.5% and 6%) on soluble recombinant matrix metalloproteinases (MMP-2, -8 and -9) and cysteine cathepsins (cathepsin B and K) were evaluated using MMP and cysteine cathepsins fluorometric assay kits. Chlorhexidine (CHX) was used as an inhibitor control. The effect of PA on endogenous matrix-bound proteases was examined by determining the change in dry mass of demineralized dentine beams and solubilized collagen peptides over 30 days. Two-way ANOVA and Tukey multiple comparison tests were used to analyze the effect of PA and proteases on the percentage inhibition of soluble proteases (α=0.05). Kruskal-Wallis one-way ANOVA and Dunn's multiple comparison tests were used to analyse the effect of PA on loss of dry mass and hydroxyproline content over time (α=0.05). RESULTS:Proanthocyanidin inactivated more than 90% of soluble recombinant MMP-2, -8 and -9 and around 75-90% of cysteinecathepsin B and K, which was significantly higher than CHX (P<0.05). The inhibition of endogenous proteases by PA increased in a dose-dependent manner. The loss of dry mass and hydroxyproline release in the medium over time was the lowest in dentine beams pretreated with PA<CHX<control (P<0.05). CONCLUSION:Proanthocyanidin exhibited both dentine MMP and cysteine cathepsins inhibition, which was higher than chlorhexidine.
Authors: Ana Beatriz Silva Sousa; Cristina M P Vidal; Ariene Arcas Leme-Kraus; Fernanda C P Pires-de-Souza; Ana K Bedran-Russo Journal: Dent Mater Date: 2016-08-11 Impact factor: 5.304
Authors: Mackeler Ramos Polassi; Thales de Sá Oliveira; Ana Calheiros de Carvalho; Lívia Soman de Medeiros Medeiros; Thiago André Moura Veiga; Carlos Frederico de Oliveira Graeff; Alejandra Hortencia Miranda González; Maria Cristina Marcucci; Simone Dos Santos Grecco; Paulo Henrique Perlatti D'Alpino Journal: ScientificWorldJournal Date: 2021-06-15