Limits...
Grape seed extracts inhibit dentin matrix degradation by MMP-3.

Khaddam M, Salmon B, Le Denmat D, Tjaderhane L, Menashi S, Chaussain C, Rochefort GY, Boukpessi T - Front Physiol (2014)

Bottom Line: Grape seed extracts (GSE) have been previously reported to be natural inhibitors of MMPs.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).However, the procedure should be adapted to clinically relevant durations.

View Article: PubMed Central - PubMed

Affiliation: EA 2496, Orofacial Pathologies, Imaging and Biotherapies, Dental school, University Paris Descartes, Sorbonne Paris Cité Montrouge, France.

ABSTRACT

Unlabelled: 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.

No MeSH data available.


Related in: MedlinePlus

Quantification of NCP release from Western blot analysis. Quantification of the different bands by ImageJ after normalization with placebo bands shows a significant weaker protein release in samples pre-treated with AmF (all proteins tested) and PBS (BGN), and a significant stronger release in NaF (BGN and DCN) and PBS (DSP and DCN) pre-treated groups. Black bars: DSP; gray bars: BGN; white bars: DCN. *p < 0.05 vs. placebo value.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4215787&req=5

Figure 2: Quantification of NCP release from Western blot analysis. Quantification of the different bands by ImageJ after normalization with placebo bands shows a significant weaker protein release in samples pre-treated with AmF (all proteins tested) and PBS (BGN), and a significant stronger release in NaF (BGN and DCN) and PBS (DSP and DCN) pre-treated groups. Black bars: DSP; gray bars: BGN; white bars: DCN. *p < 0.05 vs. placebo value.

Mentions: Western blot analysis of supernatants retrieved from the samples pre-treated with mouthrinse and GSE, followed by MMP-3 treatment, did not reveal any detectable DCN band (Figure 1). In contrast, DCN was visualized as a large band around 60 kDa in the AmF, placebo, NaF, and PBS pre-treated groups, and as an additional weaker band at 130 kDa mainly detectable in the AmF and placebo pre-treated group (Figure 1A). BGN was detected as a single large band around 140 kDa in the supernatants of the samples pre-treated with AmF, NaF, placebo, and PBS, but not with the mouthrinse and GSE (Figure 1B). For both SLRPs, no band was detected in the CHX and ZnCl2 pre-treated groups (Figures 1A,B). DSP was also released by MMP-3 and was observed as two bands around 150 and 75 kDa in the AmF, placebo, NaF and PBS pre-treated groups. No signal was detected in the mouthrinse, GSE and ZnCl2 supernatants, whereas a smear labeling was seen with CHX pre-treatment (Figure 1C). Quantification of the different bands after normalization with placebo is showed in Figure 2. In particular, release of DSP after pre-treatment with AmF was around 0.6 (p < 0.05), compared to release of DSP in placebo (set at 1), and release of BGN and DCN after pre-treatment with AmF were both around 0.4 (p < 0.05). Contrary to this, release of BGN and DCN after NaF pre-treatment were significantly higher compared to placebo (1.6 and 1.8, respectively, p < 0.05). At last, release of BSP and DCN after pre-treatment with PBS were significantly higher compared to placebo (1.8 and 2.8, respectively, p < 0.05) whereas release of BGN was significantly lower compared to placebo (0.9, p < 0.05). For MEPE, no signal was detected (data not shown).


Grape seed extracts inhibit dentin matrix degradation by MMP-3.

Khaddam M, Salmon B, Le Denmat D, Tjaderhane L, Menashi S, Chaussain C, Rochefort GY, Boukpessi T - Front Physiol (2014)

Quantification of NCP release from Western blot analysis. Quantification of the different bands by ImageJ after normalization with placebo bands shows a significant weaker protein release in samples pre-treated with AmF (all proteins tested) and PBS (BGN), and a significant stronger release in NaF (BGN and DCN) and PBS (DSP and DCN) pre-treated groups. Black bars: DSP; gray bars: BGN; white bars: DCN. *p < 0.05 vs. placebo value.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215787&req=5

Figure 2: Quantification of NCP release from Western blot analysis. Quantification of the different bands by ImageJ after normalization with placebo bands shows a significant weaker protein release in samples pre-treated with AmF (all proteins tested) and PBS (BGN), and a significant stronger release in NaF (BGN and DCN) and PBS (DSP and DCN) pre-treated groups. Black bars: DSP; gray bars: BGN; white bars: DCN. *p < 0.05 vs. placebo value.
Mentions: Western blot analysis of supernatants retrieved from the samples pre-treated with mouthrinse and GSE, followed by MMP-3 treatment, did not reveal any detectable DCN band (Figure 1). In contrast, DCN was visualized as a large band around 60 kDa in the AmF, placebo, NaF, and PBS pre-treated groups, and as an additional weaker band at 130 kDa mainly detectable in the AmF and placebo pre-treated group (Figure 1A). BGN was detected as a single large band around 140 kDa in the supernatants of the samples pre-treated with AmF, NaF, placebo, and PBS, but not with the mouthrinse and GSE (Figure 1B). For both SLRPs, no band was detected in the CHX and ZnCl2 pre-treated groups (Figures 1A,B). DSP was also released by MMP-3 and was observed as two bands around 150 and 75 kDa in the AmF, placebo, NaF and PBS pre-treated groups. No signal was detected in the mouthrinse, GSE and ZnCl2 supernatants, whereas a smear labeling was seen with CHX pre-treatment (Figure 1C). Quantification of the different bands after normalization with placebo is showed in Figure 2. In particular, release of DSP after pre-treatment with AmF was around 0.6 (p < 0.05), compared to release of DSP in placebo (set at 1), and release of BGN and DCN after pre-treatment with AmF were both around 0.4 (p < 0.05). Contrary to this, release of BGN and DCN after NaF pre-treatment were significantly higher compared to placebo (1.6 and 1.8, respectively, p < 0.05). At last, release of BSP and DCN after pre-treatment with PBS were significantly higher compared to placebo (1.8 and 2.8, respectively, p < 0.05) whereas release of BGN was significantly lower compared to placebo (0.9, p < 0.05). For MEPE, no signal was detected (data not shown).

Bottom Line: Grape seed extracts (GSE) have been previously reported to be natural inhibitors of MMPs.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).However, the procedure should be adapted to clinically relevant durations.

View Article: PubMed Central - PubMed

Affiliation: EA 2496, Orofacial Pathologies, Imaging and Biotherapies, Dental school, University Paris Descartes, Sorbonne Paris Cité Montrouge, France.

ABSTRACT

Unlabelled: 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.

No MeSH data available.


Related in: MedlinePlus