Grape seed extracts inhibit dentin matrix degradation by MMP-3

Abstract

Since Matrix metalloproteinases (MMPs) have been suggested to contribute to dentin caries progression, the hypothesis that MMP inhibition would affect the progression of dentin caries is clinically relevant. Grape seed extracts (GSE) have been previously reported to be natural inhibitors of MMPs.Objective: To evaluate the capacity of a GSE mouthrinse to prevent the degradation of demineralized dentin matrix by MMP-3 (stromelysin-1).Materials and Methods: Standardized blocks of dentin obtained from sound permanent teeth extracted for orthodontic reasons were demineralized with Ethylenediaminetetraacetic acid (EDTA) and pretreated either with (A) GSE (0.2% w/v), (B) amine fluoride (AmF) (20% w/v), (C) a mouthrinse which contains both, (D) placebo, (E) sodium fluoride (0.15 mg.ml−1), (F) PBS, (G) Chlorhexidine digluconate (CHX), or (H) zinc chloride (ZnCl2). The dentin blocks were then incubated with activated recombinant MMP-3. The supernatants were analyzed by Western Blot for several dentin matrix proteins known to be MMP-3 substrate. In parallel, scanning electron microscopy (SEM) was performed on resin replica of the dentin blocks.Results: Western blot analysis of the supernatants revealed that MMP-3 released from the dentin matrix small proteoglycans (decorin and biglycan) and dentin sialoprotein (DSP) in the AmF, sodium fluoride, PBS and placebo pretreated groups, but not in the GSE and mouthrinse pretreated groups. SEM examination of resin replica showed that the mouthrinse and its active components not only had an anti-MMP action but also modified the dentin surface accessibility.Conclusion: This study shows that GSE either alone or combined with AmF as in the evaluated mouthrinse limits dentin matrix degradation. This association may be promising to prevent the progression of caries within dentin. However, the procedure should be adapted to clinically relevant durations.

Matrix Metalloproteinases and Other Matrix Proteinases in Relation to Cariology: The Era of ‘Dentin Degradomics'

“…Suppa et al [2006] demonstrated reduced proteoglycan antigenicity together with loss of collagen antigenicity in caries-affected dentin. MMP-3, present in dentin [Boukpessi et al, 2008;Mazzoni et al, 2011] and known to degrade proteoglycans, has been shown to extract the proteoglycans decorin and biglycan and selectively also some phosphorylated proteins, whose presence might be important for remineralization [Boukpessi et al, 2008;Khaddam et al, 2014]. Inhibition of external MMP-3 by grape seed extracts totally and by fluorides partially abolished the release of decorin, biglycan and dentin sialoprotein from human dentin [Khaddam et al, 2014].…”

“…MMP-3, present in dentin [Boukpessi et al, 2008;Mazzoni et al, 2011] and known to degrade proteoglycans, has been shown to extract the proteoglycans decorin and biglycan and selectively also some phosphorylated proteins, whose presence might be important for remineralization [Boukpessi et al, 2008;Khaddam et al, 2014]. Inhibition of external MMP-3 by grape seed extracts totally and by fluorides partially abolished the release of decorin, biglycan and dentin sialoprotein from human dentin [Khaddam et al, 2014]. As papain gel, also known to extract proteoglycans, decreases the mechanical properties of demineralized dentin collagen and might even affect the collagen fibril structure ], MMP-3 might contribute to the loss of carious dentin matrix mechanical properties and reduce the ability for natural remineralization.…”

Role of Host-Derived Proteinases in Dentine Caries and Erosion

“…In addition to the reduced matrix degradation, proanthocyanidins may affect the preservation of nucleation sites for remineralization. GSE completely and AmF partially inhibit the MMP-3-induced liberation of decorin, biglycan and dentine sialoprotein, while NaF does not [Khaddam et al, 2014]. In vitro studies have also shown surface remineralization of artificial dentine caries lesions, the effect being comparable to fluoride [Xie et al, 2008;Tang et al, 2013].…”

“…Interestingly, a concentration-dependent increase in demineralized dentine elastic modulus and surface mineral deposition was seen when CHXtreated dentine was incubated in simulated body fluid [Kim et al, 2011], indicating that enzyme inhibition may improve dentine collagen remineralization. CHX also inhibits release of the non-collagenous proteins decorin, biglycan and dentine sialoprotein from the demineralized dentine matrix by MMP-3 [Khaddam et al, 2014], which might contribute to the remineralization seen with CHX. Partial remineralization (especially at the lesion surface) but deeper lesions were observed in the fluoride group, while nearly complete remineralization was seen in the fluoride-CHX group in a biofilm model [Deng et al, 2005].…”

“…In addition, the role of stromelysins (MMP-3) and the presence or absence of dentine non-collagenous proteins are not well known. MMP-3 can release small leucine-rich proteoglycans and small integrin-binding ligand N-linked glycoproteins from dentine [Chaussain et al, 2013;Khaddam et al, 2014]. These proteins participate in dentine mineralization [Chaussain et al, 2013], and immobilized phosphorylated proteins induce mineral formation in vitro and in vivo [Lussi and Linde, 1993;Saito et al, 1998].…”