Initial Enamel Erosion Research Articles

Effect of combining aminomethacrylate and fluoride against erosive and abrasive challenges on enamel and dentin.

This study evaluated the effect of solutions containing aminomethacrylate copolymer (AA) and sodium fluoride (F; 225ppm F-) or fluoride plus stannous chloride (FSn; 225ppm F-, 800ppm Sn2+) against enamel and dentin erosion/abrasion. Solutions F, FSn, AA, F+AA, FSn+AA, and deionized water as negative control were tested. Bovine enamel and dentin specimens (n=13/solution/substrate) underwent a set of erosion-abrasion cycles (0.3% citric acid [5min, 4×/day], human saliva [1h, 4×/day], brushing [15 s, 2×/day], and treatments [2min, 2×/day]) for each of five days. Initial enamel erosion was evaluated using Knoop microhardness after the first and second acid challenge on day 1, and surface loss with profilometry after day 5. KOH-soluble fluoride was assessed. Data were analyzed with ANOVA/Tukey tests. The combination of fluoride and AA resulted in higher protection against enamel erosion, whereas this was not the case for the combination of AA and FSn. All treatments protected against enamel and dentin loss. The lowest surface loss values were observed with F+AA and FSn+AA. The polymer did not significantly influence the KOH-soluble fluoride formation on enamel/dentin specimens. The aminomethacrylate copolymer effectively enhanced the efficacy of sodium fluoride against initial erosion and improved the control of enamel and dentin wear of F and FSn solutions.

The effect of toothpaste with reduced concentration of fluoride-containing sodium trimetaphosphate and polyols on initial enamel erosion.

This in vitro study evaluated the efficacy of toothpaste containing fluoride (F), sodium trimetaphosphate (TMP), and xylitol and erythritol (XE) to inhibit or repair initial enamel erosion lesions. Bovine enamel blocks (n = 120) were selected according to surface hardness (SH) and randomly divided into 5 experimental groups (n = 24 blocks/group): Placebo (no F, TMP, XE); 1100ppm F; 16% xylitol + 4% erythritol (XE); 200ppm F + 0.2% TMP (200ppm F/TMP); and 200ppm F + 0.2% TMP + 16% xylitol + 4% erythritol (200ppm F/TMP/XE). The sound and softened blocks were immersed in toothpaste slurry in human saliva for 2min. The blocks were then submitted to 4 erosive challenges in citric acid (0.75%, pH 3.5), each challenge for 1min, with stirring. The SH of the blocks was determined after treatment (t) and after the 4 erosive challenges. In addition, the precipitates were analyzed by scanning electron microscopy (SEM). Variables were submitted to a two-way repeated-measures analysis of variance followed by a Student-Newman-Keuls test (p < 0.05). Toothpaste containing 200ppm F/TMP/XE led to the highest protective and repair effect compared to the other groups (p < 0.001). The protective and repair effect was XE > 200ppm F/TMP > 1100ppm F > placebo (p < 0.001). All groups produced precipitates with a thicker layer for XE and TMP groups. Toothpaste containing 200ppm F, TMP, and polyols demonstrated a superior protective and repair effect in initial enamel erosive lesions in vitro. Toothpaste containing F and polyols could be an advantage in patients affected by dental erosion, due to its higher preventive potential, mainly in individuals who frequently drink acidic beverages. However, future studies are needed to confirm these results.

New insights into the anti-erosive property of a sugarcane-derived cystatin: different vehicle of application and potential mechanism of action.

A new sugarcane-derived cystatin (CaneCPI-5) showed anti-erosive properties when included in solutions and strong binding force to enamel, but the performance of this protein when added to gel formulations and its effect on surface free energy (SFE) requires further studies. Objective 1) to evaluate the protective effect of gels containing different concentrations of CaneCPI-5 against initial enamel erosion (Experiment 1); and 2) to analyze the SFE (γS) after treating the enamel surface with CaneCPI-5 solution (Experiment 2).Methodology In Experiment 1, 75 bovine enamel specimens were divided into five groups according to the gel treatments: placebo (negative control); 0.27%mucin+0.5%casein (positive control); 0.1 mg/mL CaneCPI-5; 1.0 mg/mL CaneCPI-5; or 2.0 mg/mL CaneCPI-5. Specimens were treated with the gels for 1 min, the AP was formed (human saliva) for 2 h and the specimens were incubated in 0.65% citric acid (pH=3.4) for 1 min. The percentage of surface hardness change (%SHC) was estimated. In Experiment 2, measurements were performed by an automatic goniometer using three probing liquids: diiodomethane, water and ethylene glycol. Specimens (n=10/group) remained untreated (control) or were treated with solution containing 0.1 mg/mL CaneCPI-5, air-dried for 45 min, and 0.5 µL of each liquid was dispensed on the surface to measure contact angles.Results Gels containing 0.1 and 1.0 mg/mL CaneCPI-5 significantly reduced %SHC compared to the other treatments (p<0.05). Treated enamel showed significantly lower γS than control, without changes in the apolar component (γSLW), but the polar component (γSAB=Lewis acid-base) became more negative (p<0.01). Moreover, CaneCPI-5 treatment showed higher γS - (electron-donor) values compared to control (p<0.01).Conclusions Gels containing 0.1 mg/mL or 1.0 mg/mL CaneCPI-5 protected enamel against initial dental erosion. CaneCPI-5 increased the number of electron donor sites on the enamel surface, which may affect AP formation and could be a potential mechanism of action to protect from erosion.

Impact of early biofilm on the assessment of initial enamel erosion with swept-source optical coherence tomography.

Objective: This study examined the impact of early biofilm on the tooth surface, during the assessment of initial enamel erosion using swept-source optical coherence tomography (SS-OCT). Method and materials: Forty-five enamel windows of 2 × 4 mm2 were prepared on 23 extracted human teeth. The specimens were exposed to citric acid (pH 3.2) for 30 minutes and randomly divided into three groups (n = 15): Group 1, no biofilm; Group 2, 1-day-old biofilm; and Group 3, 3-day-old biofilm. Specimens in Groups 2 and 3 were inoculated with oral bacteria (Streptococcus sanguinis, Streptococcus mitis, and Actinomyces naeslundii) to produce early laboratory-cultivated biofilms for 1 and 3 days respectively. Surface microhardness (SMH) measurements were taken at pre- (t1) and post-erosion (t2); and SS-OCT scans were done at t1, t2, and post-biofilm cultivation (t3). Integrated reflectivity (IR) of the tooth-air interface (IRsurface) and enamel (IRenamel) were computed from the mean A-scans. Statistical analysis was performed using paired t tests and one-way ANOVA (α = .05). Results: A significant increase in IRenamel was observed at t2 (P < .05). At t3, IRsurface between Group 1 (control) and Group 2 (P = .012) as well as Group 3 (P = .001) were significantly different. Significant variances in IRenamel were perceived between t2 and t3 for Groups 2 and 3 but not for Group 1. Conclusion: As early biofilm affected SS-OCT assessment of initial enamel erosion, they should be removed from the tooth surface prior to OCT procedures..

Salivary Hemoglobin Protects against Erosive Tooth Wear in Gastric Reflux Patients

Objective: Saliva is the most important biological factor to protect against erosive tooth wear (ETW). Gastroesophageal reflux disease (GERD) patients have an increased risk of ETW due to the frequent presence of intrinsic acids in the oral cavity. Remarkably, not all GERD patients suffer from ETW, which might be due to differences in the composition of the saliva. Methods: This study compared the proteomic profile of saliva in patients (1) with GERD and ETW (basic erosive wear examination, BEWE, score ≥9; GE group) and (2) with GERD without ETW (BEWE = 0; GNE group) using shotgun label-free quantitative proteomic analysis nLC-ESI-MS/MS. The ability of hemoglobin (Hb) to protect against initial enamel erosion caused by a daily 10-s immersion of enamel specimens in 0.01 M HCl (pH 2.3) for 3 days was evaluated in vitro for proof of concept. Surface hardness change was used as response variable. Results: The differential expression of Hb subunits was significantly increased in the GNE group versus the GE group, in particular the Hb α-subunit that showed a >22-fold increase. Expressions of serum albumin (4.5-fold) and isoforms of cytoskeletal keratin type II (>3-fold) were also increased in the GNE group. Proteinase inhibitors, such as α₁-antitrypsin and α₂-macroglobulin, were only identified in the GNE group. In vitro, Hb (1.0 and 4.0 mg/mL) significantly reduced initial enamel erosion compared to a negative control after 3 days. Conclusions: Our results indicate that many proteins, with special emphasis on Hb, may be involved in the resistance of GERD patients to the occurrence of ETW. These proteins may be candidates for inclusion in dental products to protect against ETW.

Statherin-derived peptide protects against intrinsic erosion

ObjectivesIn the present study, we used an in vitro initial intrinsic erosion model to evaluate: (experiment 1) the influence of the degree of serine (Ser) phosphorylation of peptides containing the 15 N-terminal residues of statherin and (experiment 2) the effect of different concentrations of the peptide with the best performance in experiment 1 on initial enamel erosion. DesignBovine enamel specimens were divided into 6 groups (n = 15/group) for each experiment. In experiment 1, the peptides evaluated (at 1.88 × 10−5 M) were: not phosphorylated (StatSS), phosphorylated in Ser2 (StatpSS), phosphorylated in Ser3 (StatSpS) phosphorylated in Ser2 and Ser3 (StatpSpS). Phosphate buffer and human recombinant statherin were used as negative and positive controls, respectively. In experiment 2, StatpSpS was evaluated at different concentrations: 0.94, 1.88, 3.76 and 7.52 × 10−5 M. Phosphate buffer and 0.1 mg/mL CaneCPI-5 were employed as negative and positive controls, respectively. In each experiment, the specimens were incubated with the solutions for 2 h, then the AEP was allowed to form (under human pooled saliva) for 2 h. The specimens were then challenged with 0.01 M HCl for 10 s. Demineralization was evaluated by percentage of surface hardness change (%SHC). Data were analyzed by ANOVA and Tukey’s test (p < 0.05). ResultsIn experiment 1, only StatpSpS significantly reduced the % SHC in comparison with control. In experiment 2, 1.88 × 10−5 M StatpSpS significantly reduced the %SHC in comparison with control. ConclusionsThis is the first study showing that statherin-derived peptide might protect against intrinsic erosion.

Chemical and physical factors of desensitizing and/or anti-erosive toothpastes associated with lower erosive tooth wear

Toothpastes have a complex formulation and their different chemical and physical factors will influence their effectiveness against erosive tooth wear (ETW). We, therefore, investigated the effect of different desensitizing and/or anti-erosive toothpastes on initial enamel erosion and abrasion, and analysed how the interplay of their chemical and physical factors influences ETW. Human enamel specimens were submitted to 5 erosion-abrasion cycles using 9 different toothpastes and an artificial saliva group, and enamel surface loss (SL) was calculated. Chemical and physical factors (pH; presence of tin; calcium, phosphate and fluoride concentrations; % weight of solid particles; wettability; and particle size) of the toothpaste slurries were then analysed and associated with the amount of SL in a multivariate model. We observed that all desensitizing and/or anti-erosive toothpastes presented different degrees of SL. Besides pH and fluoride, all other chemical and physical factors were associated with SL. The results of this experiment indicate that enamel SL occurs independent of whether the toothpastes have a desensitizing or anti-erosive claim, and that lower SL is associated with the presence of tin, higher concentration of calcium and phosphate, higher % weight of solid particles, smaller particle size, and lower wettability.

A New Sugarcane Cystatin Strongly Binds to Dental Enamel and Reduces Erosion

Cystatin B was recently identified as an acid-resistant protein in acquired enamel pellicle; it could therefore be included in oral products to protect against caries and erosion. However, human recombinant cystatin is very expensive, and alternatives to its use are necessary. Phytocystatins are reversible inhibitors of cysteine peptidases that are found naturally in plants. In plants, they have several biological and physiological functions, such as the regulation of endogenous processes, defense against pathogens, and response to abiotic stress. Previous studies performed by our research group have reported high inhibitory activity and potential agricultural and medical applications of several sugarcane cystatins, including CaneCPI-1, CaneCPI-2, CaneCPI-3, and CaneCPI-4. In the present study, we report the characterization of a novel sugarcane cystatin, named CaneCPI-5. This cystatin was efficiently expressed in Escherichia coli, and inhibitory assays demonstrated that it was a potent inhibitor of human cathepsins B, K, and L (Ki = 6.87, 0.49, and 0.34 nM, respectively). The ability of CaneCPI-5 to bind to dental enamel was evaluated using atomic force microscopy. Its capacity to protect against initial enamel erosion was also tested in vitro via changes in surface hardness. CaneCPI-5 showed a very large force of interaction with enamel (e.g., compared with mucin and casein) and significantly reduced initial enamel erosion. These results suggest that the inclusion of CaneCPIs in dental products might confer protection against enamel erosion.

Fluoride varnishes containing calcium glycerophosphate: fluoride uptake and the effect on in vitro enamel erosion.

Calcium glycerophosphate (CaGP) was added to fluoride varnishes to analyze their preventive effect on initial enamel erosion and fluoride uptake: potassium hydroxide (KOH)-soluble and KOH-insoluble fluoride bound to enamel. This study was carried out in two parts. Part 1: 108 enamel samples were randomly distributed into six varnish groups: base varnish (no active ingredients); Duraphat® (2.26%NaF); Duofluorid® (5.63%NaF/CaF2); experimental varnish 1 (1%CaGP/5.63 NaF/CaF2); experimental varnish 2 (5%CaGP/5.63%NaF/CaF2); and no varnish. Cyclic demineralization (90 s; citric acid, pH = 3.6) and remineralization (4 h) was made once a day, for 3 days. Change in surface microhardness (SMH) was measured. Part 2: 60 enamel samples were cut in half and received no varnish (control) or a layer of varnish: Duraphat®, Duofluorid®, experimental varnishes 1 and 2. Then, KOH-soluble and KOH-insoluble fluoride were analyzed using an electrode. After cyclic demineralization, SMH decreased in all samples, but Duraphat® caused less hardness loss. No difference was observed between varnishes containing CaGP and the other varnishes. Similar amounts of KOH-soluble and insoluble fluoride was found in experimental varnish 1 and Duofluorid®, while lower values were found for experimental varnish 2 and Duraphat®. The addition of CaGP to fluoride varnishes did not increase fluoride bound to enamel and did not enhance their protection against initial enamel erosion. We observe that the fluoride varnishes containing CaGP do not promote greater amounts of fluoride bound to enamel and that fluoride bound to enamel may not be closely related to erosion prevention.

Measuring Initial Enamel Erosion with Quantitative Light-Induced Fluorescence and Optical Coherence Tomography: An in vitro Validation Study

Background: Measurement of initial enamel erosion is currently limited to in vitro methods. Optical coherence tomography (OCT) and quantitative light-induced fluorescence (QLF) have been used clinically to study advanced erosion. Little is known about their potential on initial enamel erosion. Objectives: To evaluate the sensitivity of QLF and OCT in detecting initial dental erosion in vitro. Methods: 12 human incisors were embedded in resin except for a window on the buccal surface. Bonding agent was applied to half of the window, creating an exposed and non-exposed area. Baseline measurements were taken with QLF, OCT and surface microhardness. Samples were immersed in orange juice for 60 min and measurements taken stepwise every 10 min. QLF was used to compare the loss of fluorescence between the two areas. The OCT system, OCS1300SS (Thorlabs Ltd.), was used to record the intensity of backscattered light of both areas. Multiple linear regression and paired t test were used to compare the change of the outcome measures. Results: All 3 instruments demonstrated significant dose responses with the erosive challenge interval (p < 0.05) and a detection threshold of 10 min from baseline. Thereafter, surface microhardness demonstrated significant changes after every 10 min of erosion, QLF at 4 erosive intervals (20, 40, 50 and 60 min) while OCT at only 2 (50 and 60 min). Conclusion: It can be concluded that OCT and QLF were able to detect demineralization after 10 min of erosive challenge and could be used to monitor the progression of demineralization of initial enamel erosion in vitro.

Combined effect of a fluoride-, stannous- and chitosan-containing toothpaste and stannous-containing rinse on the prevention of initial enamel erosion–abrasion

ObjectivesThe aim of this study was to assess the preventive effect of a fluoride-, stannous- and chitosan-containing (F/Sn/chitosan-) toothpaste (TP) on initial enamel erosion and abrasion. MethodsIn total, 150 human premolar enamel specimens were ground, polished and divided into 5 toothpaste/rinse groups (n=30): (G1) placebo-TP/tap water, (G2) sodium fluoride (NaF-) TP/tap water, (G3) F/Sn/chitosan-TP/tap water, (G4) F/Sn/chitosan-TP/Sn-rinse, (G5) NaF-TP/NaF-rinse. The 8-day erosion–abrasion cyclic treatment (one cycle/day) consisted of incubating the samples in artificial saliva (30min), then submitting the samples to toothbrush abrasion (2min incubation in toothpaste slurry; brushing with 20 toothbrush strokes) and rinsing (2min; 10ml) with the respective solution: tap water (G1–G3), Sn-rinse (G4) or NaF-rinse (G5). Afterwards, the samples were submitted to erosion (2min; 30ml 1% citric acid, pH=3.6). Surface microhardness (SMH) was measured initially and after every abrasion and erosion treatment. Enamel substance loss was calculated after each abrasion. Non-parametric ANOVA followed by Wilcoxon rank tests were used for analysis. ResultsG1 presented the greatest SMH decrease, while G4 presented the least SMH decrease (p<0.001). G3 had a similar SMH decrease to G2 and G5. Substance loss was significantly lower in G4 than all other groups (p<0.05), closely followed by G3. Both G2 and G5 showed similar calculated enamel substance loss to G1. ConclusionThe treatment with F/Sn/chitosan-TP and tap water provided a similar SMH decrease to both NaF-TP groups, but significantly lower substance loss. F/Sn/Chitosan-TP and Sn-rinse showed a better preventive effect, which promoted less SMH decrease and reduced substance loss. Clinical significanceThe toothpaste containing fluoride, stannous and chitosan shows promising results in reducing substance loss from erosion and abrasion. The combination of this toothpaste with the stannous-containing rinse showed even better prevention against erosion–abrasion.

In vitro assessment of artificial saliva formulations on initial enamel erosion remineralization

ObjectivesVarious formulations of artificial saliva are present in the literature and little guidance is available on the standardization of type of saliva for use in in vitro protocols for erosive studies. The aim of this study was to evaluate the remineralizing capacity of different formulations of artificial saliva on initial enamel erosive lesion. MethodsBovine enamel blocks were subjected to short-term acidic exposure by immersion in citric acid 0.05M (pH 2.5) for 15s, resulting in surface softening without tissue loss. Then 90 selected eroded enamel blocks were randomly and equally divided into 6 groups according to saliva formulation (n=15): Saliva 1 (contain mucin); Saliva 2 (Saliva 1 without mucin); Saliva 3; Saliva 4; Saliva 5 (contain sodium carboxymethyl cellulose) and control (C) (deionized water). After demineralization enamel blocks were subjected to remineralization by immersion in the saliva's formulations for 2h. Enamel remineralization was measured by superficial hardness test (% superficial hardness change). The data were tested using ANOVA and Tukey's test (p<0.05). ResultsAll the tested formulations of artificial saliva resulted in significantly higher enamel remineralization compared to control (p<0.001). Saliva 3 showed higher percentage of enamel remineralization than Saliva 5 (p<0.05). ConclusionsBesides the variety of artificial saliva for erosion in vitro protocols, all the formulations tested were able to partially remineralize initial erosive lesions.