Effects of protease inhibitors on dentin erosion: an in situ study.

OBJECTIVE: This in situ study aimed to evaluate the effects of the inhibitors of matrix metalloproteinases (MMPs) and cysteine cathepsins on dentin erosion. MATERIALS AND METHODS: Ten volunteers participated in this study. Each volunteer wore an intraoral appliance containing 4 dentin specimens subjected to different treatments: deionized water as a control, 1 mM 1,10-phenanthroline (an MMP inhibitor), 50 µM E-64 (a cysteine cathepsin inhibitor), and 1 mM 1,10-phenanthroline + 50 µM E-64. The specimens were dipped in 5 ml of the respective solutions for 30 min at room temperature and then exposed to in vivo erosive challenges by rinsing with 150 ml of a cola drink (4 × 5 min/day) for 7 days. The substance loss of the specimens was measured by profilometry. The transverse sections of the specimens were examined using scanning electron microscopy. Thereafter, the demineralized organic matrix (DOM) of the specimens was removed using type I collagen enzyme and assessed by performing profilometry. The differences in substance loss and DOM thickness among the groups were analyzed by one-way repeated-measures analysis of variance (ANOVA) and Bonferroni's test at a level of P < 0.05. RESULTS: Protease inhibitors significantly reduced substance loss in comparison to that of the control group (all P < 0.05). A significantly thicker DOM was observed for the specimens treated with protease inhibitors than for the control specimens (all P < 0.05). No significant differences in substance loss or DOM thickness were found among the MMP inhibitor, cysteine cathepsin inhibitor, and MMP + cysteine cathepsin inhibitor groups. CONCLUSIONS: The use of MMP and cysteine cathepsin inhibitors was shown to increase the acid resistance of human dentin, which may be due to the preservation of the DOM. CLINICAL RELEVANCE: The application of protease inhibitors could be considered an appropriate preventive strategy for dentin erosion.

Does delayed toothbrushing after the consumption of erosive foodstuffs or beverages decrease erosive tooth wear? A systematic review and meta-analysis.

OBJECTIVE: Controversy exists regarding the effectiveness of delayed toothbrushing in decreasing erosive tooth wear (ETW). The purpose of this systematic review and meta-analysis was to assess the effects of delayed toothbrushing on ETW. MATERIALS AND METHODS: This systematic review and meta-analysis was conducted according to the PRISMA statement and registered in PROSPERO (CRD42020200463). PubMed, Embase, and Web of Science databases were systematically searched with no publication year limits. Screening and data extraction were performed independently by two reviewers. In situ and in vitro studies comparing ETW after delayed and immediate toothbrushing following an erosive attack were included. Review Manager software 5.3 (The Cochrane Collaboration, Oxford, UK) was used for statistical analyses. Heterogeneity was assessed with the Cochran Q test and I2 statistics. RESULTS: Of the 565 potentially relevant studies, 26 full-text articles were assessed for eligibility. Twelve articles were included in the systematic review, and 11 were included in the qualitative analyses. No significant difference in the ETW of human enamel was observed between delayed and immediate toothbrushing (P = 0.13), whereas significantly less ETW of bovine enamel was observed after delayed toothbrushing (P < 0.001). No significant difference in the ETW of bovine dentin was observed between delayed and immediate toothbrushing (P = 0.34). Studies on human dentin were not available. Subgroup analyses revealed a significant contribution of the use of fluoridated toothpaste to decreasing the ETW of human enamel after erosion and toothbrush abrasion (P = 0.02). CONCLUSIONS: Bovine and human teeth behaved differently in response to erosion and toothbrush abrasion. Delayed toothbrushing after an erosive attack was not effective at decreasing the ETW of human enamel compared to immediate toothbrushing, whereas it was effective at decreasing the ETW of bovine enamel. CLINICAL RELEVANCE: Delayed toothbrushing alone after the consumption of erosive foodstuffs or beverages is not capable of preventing erosive enamel wear.

Dual function of quercetin as an MMP inhibitor and crosslinker in preventing dentin erosion and abrasion: An in situ/in vivo study.

OBJECTIVE: The aim of the present study was to evaluate the in situ/in vivo effect of quercetin on dentin erosion and abrasion. METHODS: Human dentin blocks (2 × 2 × 2 mm) were embedded and assigned to 6 groups: 75 µg/mL, 150 µg/mL and 300 µg/mL quercetin (Q75, Q150, Q300); 120 µg/mL chlorhexidine (CHX, positive control); and deionized water and ethanol (the negative controls). The specimens were treated with the respective solutions for 2 min and then subjected to in situ/in vivo erosive/abrasive challenge for 7 d as follows: in vivo erosion 4 times a day and then in vivo toothbrush abrasion after the first and last erosive challenges of each day. Dentin loss was assessed by profilometry. An additional dentin specimen was used to evaluate the penetration depth of quercetin into dentin by tracking the spatial distribution of its characteristic Raman peak. Moreover, dentin blocks (7 × 1.7 × 0.7 mm) were used to detect the impact of quercetin on dentin-derived matrix metalloproteinase (MMP) inhibition by in situ zymography, and the inhibition percentage (%) was calculated. Additionally, the potential collagen crosslinking interactions with quercetin were detected by Raman spectroscopy, and the crosslinking degree was determined with a ninhydrin assay. Fully demineralized dentin beams (0.5 × 0.5 × 10 mm) were used to evaluate the impact of quercetin on the mechanical properties of dentin collagen fibre by the ultimate micro-tensile strength test (µUTS). The data were analysed by one-way analysis of variance and Tukey's test (α = 0.05). RESULTS: Compared to the negative controls, all treatment solutions significantly reduced dentin loss. The dentin loss of Q150 and Q300 was significantly less than that of CHX (all P < 0.05). The amount of quercetin decreased with increasing dentin depth, and the maximum penetration depth was approximately 25-30 µm. In situ zymography showed that quercetin significantly inhibited the activities of dentin-derived MMPs. The inhibitory percentages of Q75 and Q150 were significantly lower than that of CHX (all P < 0.05), but no significant difference was found between Q300 and CHX (P = 0.58). The collagen crosslinking interactions with quercetin primarily involved hydrogen bonding and the degree of crosslinking increased in a concentration-dependent manner. Statistically significant increases in µUTS values were observed for demineralized dentin beams after quercetin treatment compared with those of the control treatments (all P < 0.05). SIGNIFICANCE: This study provides the first direct evidence that quercetin could penetrate approximately 25-30 µm into dentin and further prevent dentin erosion and abrasion by inhibiting dentin-derived MMP activity as well as crosslinking collagen of the demineralized organic matrix.