Effects of Cathepsin K Inhibitors on Dentin Erosion: An in vitro Study.

Cathepsin K (catK) modulates the degradation of dentin collagen. This study aimed to evaluate the effects of catK inhibitors on dentin erosion. Dentin beams were eroded (4 times/d for 5 days) and immersed in deionized water (negative control), 0.1 M NaCl, 0.3 M NaCl, 0.5 M NaCl, or 1 µm odanacatib (each n = 16) for 30 min after each erosive challenge. Erosive dentin loss (EDL) and demineralized organic matrix (DOM) thickness were evaluated profilometrically. Additionally, dentin beams were demineralized, immersed in the respective solutions for 30 min each (n = 5), and then incubated in artificial saliva for 5 days. Dentin collage degradation was evaluated by quantifying the levels of the C-terminal peptide of type I collagen (CTX), C-terminal cross-linked telopeptide of type I collagen (ICTP), and hydroxyproline (HYP) in the incubation media. Significantly lower EDL and dentin collagen degradation (CTX, ICTP, and HYP) and thicker DOM layers were observed in the samples treated with 0.3 m NaCl and 1 µm odanacatib than in those treated with deionized water (all p < 0.05). The samples treated with 1 µm odanacatib showed significantly lower levels of CTX and HYP than those treated with 0.3 M NaCl (all p < 0.05). The present findings support the potential use of catK inhibitors in controlling dentin erosion.

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.

Effects of the application timing of anti-erosive agents on dentin erosion.

This in vitro study aimed to evaluate the effects of the application timing of anti-erosive agents on dentin erosion. Eighty dentin specimens with dimensions of 2 × 2 × 2 mm were prepared and randomly divided into 4 groups based on the treatment solutions: 1.23 × 104 µg/ml sodium fluoride (NaF), 120 µg/ml chlorhexidine (CHX), 300 µg/ml quercetin (QUE), and deionized water (DW, negative control). The specimens in each group were further divided into 2 subgroups according to the application timing of the treatment solutions (n = 10): before the erosive challenges (PRE) and after the erosive challenges (POST). All specimens were submitted to 4 daily erosive challenges for 5 d. For each erosive challenge, the specimens in the subgroup PRE were treated with the respective solutions for 2 min and then immersed in cola drinks for 5 min, while the specimens in the subgroup POST were immersed in cola drinks for 5 min followed by treatment with the respective solutions for 2 min. The erosive dentin loss (EDL) was measured using a contact profilometer, and the surface morphology of the dentin specimens was evaluated by scanning electron microscopy at the end of the experiment. The data were statistically analyzed using two-way analysis of variance (ANOVA) and Bonferroni's test (α = 0.05). Significantly less EDL was observed in the groups NaF, CHX, and QUE than in the group DW (all P < 0.001). Significantly lower EDL was observed in the groups CHX and QUE than in the group NaF (P = 0.001 and P < 0.001, respectively). For CHX, subgroup POST exhibited significantly less EDL than subgroup PRE (P < 0.001). Regarding QUE, subgroup PRE showed significantly less EDL than subgroup POST (P < 0.001). Furthermore, a relatively greater number of obliterated dentinal tubules was visible in the subgroup POST rather than in the subgroup PRE of the group CHX, while in the group QUE, narrower dentinal tubules were observed in the subgroup PRE than those in subgroup POST. In conclusion, CHX and QUE showed the best performance in controlling dentin erosion. CHX was more effective in reducing EDL when applied after erosive challenges, whereas QUE worked more effectively when used before erosive attacks. The application timing should be considered when evaluating the effects of anti-erosive agents because it may determine their effectiveness.

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.

Effect of quercetin pretreatment on the immediate and aged bond strength of bleached dentin.

This in vitro study aimed to investigate the effect of quercetin pretreatment on the bond strength of bleached dentin. Human dentin blocks (2 × 2 × 1 mm) were prepared and randomly divided into 5 groups (n = 16): deionized water pretreatment + no bleaching treatment (DNB); deionized water pretreatment + bleaching treatment (DYB); 75 µg/mL quercetin pretreatment + bleaching (Q75B); 150 µg/mL quercetin pretreatment + bleaching (Q150B); and 300 µg/mL quercetin pretreatment + bleaching (Q300B). The surfaces of superficial dentin (bonding surfaces) were treated with the respective solutions for 2 min, and then the surfaces opposite to the bonding surfaces (near pulp, bleaching surfaces) were subjected to bleaching treatment with 40% hydrogen peroxide (Ultradent, USA) for two 15-min sessions (groups DYB, Q75B, Q150B, and Q300B). After the bleaching procedure, the bonding surfaces were bonded with resin cements (Panavia V5, Kuraray, Japan). The bonded specimens were then divided into 2 subgroups (n = 8): the aging group (subgroup T), which was subjected to 10,000 thermocycles, and the nonaging group (subgroup N), which was not subjected to thermocycling. The microshear bond strength (µSBS) was obtained using a universal testing machine (AGS-X, Shimadzu, Tokyo, Japan). Additional dentin blocks (5 × 5 × 1 mm) were prepared and treated the same as the groups DYB, Q75B, Q150B, and Q300B (n = 8) to evaluate the color change, defined as groups CCDYB, CCQ75B, CCQ150B, and CCQ300B, respectively. Color evaluation was performed using a spectrophotometer (Vita Easyshade Advance 4.0, Vident, USA) to obtain a baseline and again at the end of the bleaching treatment. The data were analyzed via two-way analysis of variance (ANOVA) and Tukey's post-hoc test (α = 0.05). For the immediate bond strength, the specimens in the groups Q75B, Q150B, and Q300B showed significantly higher µSBS values than those in the group DYB (all P < 0.05). No significant differences in the µSBS values were found among the groups Q75B, Q150B, Q300B, and DNB, respectively (all P > 0.05). For the aged bond strength, both the groups Q150B and Q300B exhibited significantly higher µSBS values than groups DYB and DNB (all P < 0.05), whereas no significance differences were found between groups Q150B and Q300B (P = 1.00) or between the groups DYB and DNB (P = 1.00). No significant differences were observed in the △E values among all the groups tested (P = 0.80). Therefore, the application of quercetin for 2 min prior to the bleaching procedure preserved the immediate bond strength and improved the aged bond strength of bleached dentin while maintaining the effectiveness of bleaching.

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.

Effects of remaining dentin thickness on the bond strength of bleached dentin.

BACKGROUND: The bond strength of resin composites to dentin was reported to be related to either the remaining dentin thickness (RDT) or bleaching treatment. However, information is limited regarding the effects of RDT on the bond strength of bleached dentin. The present study aimed to investigate the effects of RDT on the microshear bond strength (µSBS) of resin cement to bleached dentin. METHODS: A total of 120 dentin specimens were prepared and randomly divided into 2 groups: a bleaching group (group B) and a control group (group C). Hydrogen peroxide with a concentration of 35% (Ultradent, USA) was applied on the dentin surface for 2 × 1 d for group B, while no bleaching treatment was performed for group C. After the treatment, the specimens were finished and polished to obtain different RDTs (2, 1, and 0.5 mm) and divided into 3 groups of 20 specimens each. The bonding procedure was performed using Panavia V5 (Kuraray, Japan) with a bonding area of 0.785 mm2. For each group, half of the specimens were subjected to 5000 thermal cycles (subgroup T), while the other half did not receive thermocycling (subgroup N) (n = 10). The specimens were then subjected to the µSBS test using a universal testing machine. Data were analyzed by a three-way analysis of variance (α = 0.05). The fracture modes of the specimens were confirmed with a measuring microscope. Representative specimens with different fracture modes were observed with scanning electron microscopy (SEM). RESULTS: The µSBS values were significantly affected by bleaching treatment (p < 0.001), whereas no significant effect was observed for thermocycling (p = 0.293). In terms of RDT, a significantly different µSBS value was found among the subgroups with different RDTs in group C (p = 0.003). However, the RDT did not significantly affect the µSBS values of bleached dentin in group B (p = 0.779). The µSBS values were significantly lower in group B than in group C (p < 0.001). A higher percentage of adhesive failure was observed in group B than in group C. CONCLUSION: Based on the present findings, it can be concluded that the RDT did not affect the bond strength of resin cement to bleached dentin. CLINICAL SIGNIFICANCE: Since RDT did not affect the bond strength of resin cement to bleached dentin, bonding procedures should not be performed immediately after intracoronal bleaching, even if the dentin is planned to be removed due to a tooth preparation process.

Stress and its association with academic performance among dental undergraduate students in Fujian, China: a cross-sectional online questionnaire survey.

BACKGROUND: The aim of this study was to investigate the amount and sources of stress in dental undergraduate students in Fujian, China, and the factors associated with stress. METHODS: This cross-sectional study was conducted during the second semester of the 2017-2018 academic year at the School of Stomatology, Fujian Medical University, China. A total of 396 students were surveyed with the Dental Environment Stress Questionnaire (DES) and the Perceived Stress Scale (PSS) using an online survey system. The participants' demographic information, including sex, age, year of study, and grade point average (GPA) was also collected. One-way analysis of variance (ANOVA) was performed to compare the stress scores. Pearson correlation and multiple linear regression analyses were conducted to explore the associated factors of stress and academic performance. All statistical analyses were performed at a significance level of 5%. RESULTS: A total of 347 undergraduate students participated in the present study, for a response rate of 87.6%. There were no significant differences in the DES and PSS total scores among students of different grades and sexes. Significant differences were found in the DES "workload" and "self-efficacy beliefs" scores among students from different study years (all P < 0.05). The Multiple linear regression showed that DES and PSS scores were negatively correlated with GPA, while sex was positively correlated with GPA (all P < 0.05). Female students had significantly higher GPAs than male students. CONCLUSIONS: Dental undergraduates in Fujian, China experienced moderate levels of stress. While the amount of stress did not differ by year of study, the sources of stress did differ. Stress scores and sex were negatively correlated with academic performance.

Knowledge of and attitudes towards erosive tooth wear among students of two Chinese universities.

BACKGROUND: To assess the knowledge of and attitudes towards erosive tooth wear among dental, medical, and non-medical university students of two Chinese universities. METHODS: A questionnaire containing 15 questions on knowledge of erosive tooth wear and 10 questions on attitudes towards erosive tooth wear was designed, and its psychometric properties (reliability and validity) were analysed in a pilot study (n = 120 students). The following 3 groups of university students (n = 635) were recruited based on a convenience sampling technique and were distributed the questionnaire via an online survey system: dental students (DSs), medical students (MSs), and non-medical students (NSs). Differences in the scores between groups and genders were analysed by one-way analysis of variance (ANOVA). The Pearson correlation coefficient was used to determine the association between the knowledge and attitude scores (P < 0.05). RESULTS: The questionnaire was found to be reliable, valid and reproducible. A total of 435 students participated in this study (response rate: 69.6%). The knowledge score of the DSs (11.5 ± 3.4) was significantly higher than those of the NSs (5.5 ± 4.0) and MSs (6.1 ± 4.0) (P < 0.001). The attitude score of the DSs (45.2 ± 6.5) was significantly higher than those of the NSs (41.1 ± 6.9) and MSs (41.8 ± 6.4) (P < 0.001). The majority of DSs expressed attitudes that were more accurate and positive than those expressed by the other 2 groups. The attitude score was positively correlated with the knowledge score (r = 0.237, P < 0.001). CONCLUSIONS: Dental students had more accurate knowledge of and more positive attitudes towards erosive tooth wear than medical and non-medical students. In this population, a positive correlation was established between knowledge of and attitudes towards erosive tooth wear.