Effect of Polyphenols on the Ultrastructure of the Dentin Pellicle and Subsequent Erosion.

INTRODUCTION: Erosive tooth wear is a highly prevalent dental condition that is modified by the ever-present salivary pellicle. The aim of the present in situ study was to investigate the effect of polyphenols on the ultrastructure of the pellicle formed on dentin in situ and a subsequent erosive challenge. METHODS: The pellicle was formed on bovine dentin specimens for 3 min or 2 h in 3 subjects. After subjects rinsed with sterile water (negative control), 1% tannic acid, 1% hop extract, or tin/fluoride solution containing 800 ppm tin and 500 ppm fluoride (positive control), specimens were removed from the oral cavity. The erosive challenge was performed on half of the specimens with 1% citric acid, and all specimens were analyzed by transmission electron microscopy. Incorporation of tannic acid in the pellicle was investigated by fluorescence spectroscopy. RESULTS: Compared to the negative control, ultrastructural analyses reveal a thicker and electron-denser pellicle after application of polyphenols, in which, according to spectroscopy, tannic acid is also incorporated. Application of citric acid resulted in demineralization of dentin, but to a lesser degree when the pellicle was pretreated with a tin/fluoride solution. The pellicle was more acid-resistant than the negative control when modified with polyphenols or tin/fluoride solution. CONCLUSION: Polyphenols can have a substantial impact on the ultrastructure and acid resistance of the dentin pellicle, while the tin/fluoride solution showed explicit protection against erosive demineralization.

Enamel Fluoride Uptake Determined Using the Microbiopsy Technique and Time-of-Flight Secondary Ion Mass Spectrometry: A Pilot Study.

This study evaluated the suitability of time-of-flight secondary ion mass spectrometry (ToF-SIMS) to assess enamel fluoride uptake (EFU) in comparison with the microbiopsy technique. Enamel specimens were exposed to equimolar solutions of fluoride prepared from sodium fluoride (NaF), stannous fluoride (SnF2), or amine fluoride (AmF). EFU was quantified by both techniques on the same specimens. EFU was found to be highest for samples treated with AmF, followed by SnF2 and NaF. Both methods yielded clearly interpretable, highly correlating (r = 0.95) data. ToF-SIMS can be considered a promising alternative to the microbiopsy technique for near-surface EFU assessment.

Combined use of stannous fluoride-containing mouth rinse and toothpaste prevents enamel erosion in vitro.

OBJECTIVE: To compare the protective effect of commercial stannous-containing mouth rinses on enamel erosion in a simulated 5-day in vitro cycling model. MATERIALS AND METHODS: 81 human enamel specimens were embedded in resin blocks and divided into nine groups as follows; group 1: stannous fluoride (1000SnF2) toothpaste; groups 2,3, and 4 were the same as group 1 plus Elmex®, PerioMed™, and Meridol®, respectively, group 5: stannous fluoride (1450SnF2) toothpaste, groups 6, 7, and 8 were the same as group 5 plus Elmex®, PerioMed™, and Meridol®, respectively, group 9: negative control. An erosive challenge was induced with a 1 min hydrochloric acid (0.01 M, pH 2.2) treatment 3 times per day. Each cycle included immersing in the toothpaste slurry twice for two minutes and a one-minute rinse. The enamel slabs were immersed in artificial saliva between each erosive cycle and incubated overnight at 37 °C. Surface hardness loss and enamel loss were determined by Knoop surface hardness and non-contact profilometry, respectively. Finally, enamel surfaces were analyzed by scanning electron microscopy and X-ray energy dispersive spectroscopy (SEM/EDS). RESULTS: All three mouth rinses had similar protective effects against erosion when using adjunct with 1000 SnF2 toothpaste (p > 0.05). With 1450 SnF2 toothpaste, Elmex® presented significantly lower surface hardness loss than Meridol® (p < 0.05). The combined use of Elmex® or PerioMed™ with toothpaste provided significantly better erosion protection than toothpaste alone, either 1000 or 1450 SnF2. In addition, 1000SnF2 toothpaste adjunct with mouth rinse is comparable to 1450 SnF2 toothpaste alone in preventing enamel erosion. CONCLUSION: All three mouth rinses reduced enamel erosion. The additional use of a high concentration stannous containing mouth rinse with 1450 SnF2 toothpaste increases the protective effect against enamel erosion in vitro. CLINICAL SIGNIFICANCE: To date, no standard protocol for preventing dental erosion is available. There are three stannous-containing mouth rinses on the market; however, no study compared their efficacy or indicated whether using adjuncts with anti-erosion toothpaste provides additional benefits. This study found that adding stannous mouth rinse to twice-daily toothpaste increases erosion protection.

A randomized controlled clinical evaluation of desensitization efficacy of a newly developed toothpaste with highly stabilized SnF2.

PURPOSE: To assess the relief of dentin hypersensitivity of the new toothpaste with stabilized stannous fluoride (SnF2) versus a marketed standard fluoride toothpaste as a negative control and a marketed anhydrous SnF2 toothpaste as a positive control. METHODS: This was a single-centered, randomized, controlled, double blind, clinical trial. 96 participants with hypersensitivity were enrolled in this 4-week clinical study. Electrical stimulation and evaporative air tests were performed to evaluate the desensitization efficacy. Clinical assessments were made at baseline, and after 3 days, 1 week, 2 weeks and 4 weeks of twice-daily brushing. Additionally, the influence of Sn² ⁺ species on desensitization was evaluated using bovine dentin specimens treated with toothpaste. RESULTS: All 96 enrolled participants were randomized. 96 participants completed all evaluations. Participants had an average age (SD) of 47.0 (10.5) years; 45% of participants were female. Both SnF2 toothpastes showed superior desensitization efficacy compared to the negative control toothpaste, the conventional sodium monofluorophosphate (SMFP) toothpaste, after a week. The new stabilized SnF2 toothpaste demonstrated improved electrical stimulation benefits compared to the negative control toothpaste, with increases of 15.1% after 3 days, 34.2% after 1 week, 66.3% after 2 weeks, and 111.6% after 4 weeks. Additionally, it showed relative verbal evaluation scale (VES) benefits of 14.2% after 3 days, 37.6% after 1 week, 28.9% after 2 weeks, and 37.4% after 4 weeks. The stabilized SnF2 toothpaste exhibited desensitization properties comparable to those of a commercial anhydrous SnF2 toothpaste, which typically produces undesirable side effects in the mouth. Toothpastes containing 0.454 % SnF2 exhibited perfect occlusion of dentin tubules. CLINICAL SIGNIFICANCE: The stabilized 0.454% SnF2 toothpaste exhibited significantly greater dentin hypersensitivity relief within only a week and comparable property to commercial anhydrous SnF2 toothpaste.

Influence of Different Mouthwashes on the Efficacy of Fluoridated Dentifrices in Prevention of Enamel Erosion: An In Vitro Study.

AIM: The purpose of the current study was to evaluate the impact of three various mouthwashes on the effectiveness of fluoride dentifrices in preventing enamel erosion. MATERIALS AND METHODS: A total of 120 sound intact human premolar teeth which were extracted for orthodontic treatment were selected for the study. A 3 × 3 mm window section was positioned in the middle of the coronal surface of the tooth in order to define the study area. Each sample was placed in a solution of 1% citric acid (pH 3.5) for 10 minutes in order to produce an eroded surface. All samples were divided into two main groups (60 samples each) as follows: Group A for sodium fluoride dentifrices and group B for stannous fluoride dentifrices, again it is subdivided into: CHX: Chlohex ADS®, EO: Listerine®, CPC: Colgate® Plax (20 samples in each subgroup). After that, samples underwent the pH cycling model for 5 days. Samples were examined for surface loss using a scanning electron microscope. RESULTS: In sodium fluoride dentifrices group, before intervention, the surface loss was 3.12 ± 1.03 in CHX group, 3.08 ± 1.20 in EO group, and 3.09 ± 0.96 in CPC group. After intervention, the less surface loss found with CHX group (2.18 ± 0.84), followed by CPC (2.34 ± 0.74) and EO group (2.46 ± 0.97). In stannous fluoride dentifrices group, before intervention, the surface loss in CHX group was 3.26 ± 1.19, in EO group, it was 3.18 ± 1.31, and in CPC group, it was 3.22 ± 1.06. After intervention, the less surface loss found with CHX: group (1.90 ± 0.54), followed by CPC (2.24 ± 0.28) and EO group (2.38 ± 0.20). CONCLUSION: The present study concluded that the fluoride dentifrices' preventive effects against tooth surface loss were unaffected by a different mouthwashes with varying compositions and major constituents. In terms of erosion, fluoridated toothpaste containing stannous fluoride was found to provide better surface loss protection than sodium fluoride. CLINICAL SIGNIFICANCE: Primary prevention and the eradication of contributing causes are the greatest strategies for preventing erosion. Simultaneously, antibacterial agent in the mouthwashes may help in enhancing the effect of fluoride in the enamel, owing to their high affinity for teeth structures. Therefore, in addition to cause-related treatment, further efforts to reduce tooth tissue loss are also necessary.

The influence of biofilm maturation on fluoride's anticaries efficacy.

OBJECTIVES: (1) To explore the influence of biofilm maturation and timing of exposure on fluoride anticaries efficacy and (2) to explore biofilm recovery post-treatment. METHODS: Bovine enamel specimens were utilized in a pH cycling model (28 subgroups [n = 18]). Each subgroup received different treatments [exposure]: sodium fluoride [NaF]; stannous fluoride [SnF2]; amine fluoride [AmF]; and de-ionized water [DIW], at a specific period: early: days 1-4; middle: days 3-6; and late: days 7-10. During non-exposure periods, pH cycling included DIW instead of fluorides. Objective 1: part 1 (cycling for 4, 6, or 10 days). Part 2 (cycling for 10 days). Objective 2: early exposure: three sample collection time points (immediate, 3 days, and 6 days post-treatment); middle exposure: two sample collection time points (immediate, 4 days post-treatment). The enamel and biofilm were analyzed ([surface microhardness; mineral loss; lesion depth]; [lactate dehydrogenase enzyme activity; exopolysaccharide amount; viability]). Data were analyzed using ANOVA (p = 0.05). RESULTS: Objective 1: Early exposure to fluorides produced protective effects against lesion progression in surface microhardness and mineral loss, but not for lesion depth. Objective 2: Early exposure slowed the demineralization process. SnF2 and AmF were superior to NaF in reducing LDH and EPS values, regardless of exposure time. They also prevented biofilm recovery. CONCLUSION: Earlier exposure to SnF2 and AmF may result in less tolerant biofilm. Early fluoride treatment may produce a protective effect against demineralization. SnF2 and AmF may be the choice to treat older biofilm and prevent biofilm recovery. CLINICAL RELEVANCE: The study provides an understanding of biofilm-fluoride interaction with mature biofilm (e.g., hard-to-reach areas, orthodontic patients) and fluoride's sustainable effect hours/days after brushing.

Recent Development of Active Ingredients in Mouthwashes and Toothpastes for Periodontal Diseases.

Periodontal diseases like gingivitis and periodontitis are primarily caused by dental plaque. Several antiplaque and anti-microbial agents have been successfully incorporated into toothpastes and mouthwashes to control plaque biofilms and to prevent and treat gingivitis and periodontitis. The aim of this article was to review recent developments in the antiplaque, anti-gingivitis, and anti-periodontitis properties of some common compounds in toothpastes and mouthwashes by evaluating basic and clinical studies, especially the ones published in the past five years. The common active ingredients in toothpastes and mouthwashes included in this review are chlorhexidine, cetylpyridinium chloride, sodium fluoride, stannous fluoride, stannous chloride, zinc oxide, zinc chloride, and two herbs-licorice and curcumin. We believe this comprehensive review will provide useful up-to-date information for dental care professionals and the general public regarding the major oral care products on the market that are in daily use.

Gingivitis efficacy of a 0.454% w/w stannous fluoride dentifrice: a 24-week randomized controlled trial.

BACKGROUND: Plaque-induced gingivitis can be prevented and treated with regular effective oral hygiene, principally via mechanical cleaning with regular toothbrushing. To complement the mechanical plaque removal, antimicrobial ingredients can be incorporated into dentifrices to inhibit the growth of plaque. This study aimed to evaluate and compare gingivitis and the proportion of subjects moving between gingivitis severity (< 10, > 10 < 30, > 30% bleeding sites), and plaque reduction, following twice daily use of an experimental non-aqueous 0.454% weight/weight (w/w) stannous fluoride (SnF2) dentifrice, compared to a negative control dentifrice over 12 and 24 weeks. METHOD: This was a single-center, examiner-blinded, randomized, stratified, two-treatment arm, parallel group, 24-week clinical study in healthy adult volunteers with moderate gingivitis. At baseline, after abstaining from toothbrushing overnight, subjects underwent MGI (modified gingival index), BI (bleeding index) and PI (plaque index) assessments. Eligible subjects, who met the inclusion/exclusion criteria, were stratified based on gender and baseline mean MGI score (Low ≤2.00 /High > 2.00) and randomized to treatment. Following randomization, subjects underwent a thorough dental prophylaxis and flossing. After 12 and 24 weeks of twice daily brushing with their allocated treatment, subjects returned to the site (with overnight plaque, having abstained from oral hygiene procedures for 8 h prior to visit) for MGI, BI and PI assessments. Treatment effect was evaluated by comparing the MGI, BI and PI scores. RESULTS: One hundred and twenty-nine subjects were screened; 98 subjects were randomized and 90 subjects completed the study. Statistically significant differences between treatments, in favour of the 0.454% stannous fluoride dentifrice were observed, compared to the negative control dentifrice, for all outcome measures (MGI, BI, bleeding sites and PI at weeks 12 and 24 p < 0.0001). At 24 weeks, 71% of subjects in the 0.453% SnF2 treatment group demonstrated < 10% of bleeding sites. CONCLUSION: A dentifrice containing 0.454% w/w SnF2 was shown to be superior to a standard dentifrice in controlling gingivitis and supra-gingival plaque, over a 24-week period. Over two thirds of subjects in the 0.454% SnF2 treatment group demonstrated a level of bleeding sites potentially representative of "clinical periodontal health" (< 10%) following a dental prophylaxis and 24 weeks of product use. TRIAL REGISTRATION: This study was retrospectively registered at ClinicalTrials.gov, on 11th Oct. 2019 (NCT04123665).

Introducing BAIT (Biofilm Architecture Inference Tool): a software program to evaluate the architecture of oral multi-species biofilms.

Biofilm model systems are used to study biofilm growth and predict the effects of anti-biofilm interventions within the human oral cavity. Many in vitro biofilm model systems use a confocal laser scanning microscope (CLSM) in conjunction with image analysis tools to study biofilms. The aim of this study was to evaluate an in-house developed image analysis software program that we call BAIT (Biofilm Architecture Inference Tool) to quantify the architecture of oral multi-species biofilms following anti-biofilm interventions using a microfluidic biofilm system. Differences in architecture were compared between untreated biofilms and those treated with water (negative control), sodium gluconate ('placebo') or stannous fluoride (SnF2). The microfluidic system was inoculated with pooled human saliva and biofilms were developed over 22 h in filter-sterilized 25 % pooled human saliva. During this period, biofilms were treated with water, sodium gluconate, or SnF2 (1000, 3439 or 10 000 p.p.m. Sn2+) 8 and 18 h post-inoculation. After 22 h of growth, biofilms were stained with LIVE/DEAD stain, and imaged by CLSM. BAIT was used to calculate biofilm biovolume, total number of objects, surface area, fluffiness, connectivity, convex hull porosity and viability. Image analysis showed oral biofilm architecture was significantly altered by 3439 and 10 000 p.p.m. Sn2+ treatment regimens, resulting in decreased biovolume, surface area, number of objects and connectivity, while fluffiness increased (P<0.01). In conclusion, BAIT was shown to be able to measure the changes in biofilm architecture and detects possible antimicrobial and anti-biofilm effects of candidate agents.

No differences in microbiome changes between anti-adhesive and antibacterial ingredients in toothpastes during periodontal therapy.

AIM: This subgroup analysis of a 12-week randomized, double-blind, and two-center trial aimed to evaluate whether two different toothpaste formulations can differentially modulate the dental microbiome. MATERIAL AND METHODS: Forty one mild to moderate periodontitis patients used as an adjunct to periodontal treatment either a toothpaste with anti-adhesive zinc-substituted carbonated hydroxyapatite (HA) or with antimicrobial and anti-adhesive amine fluoride/stannous fluoride (AmF/SnF2 ) during a 12-week period. Plaque samples from buccal/lingual, interproximal, and subgingival sites were taken at baseline, 4 weeks after oral hygiene phase, and 8 weeks after periodontal therapy. Samples were analyzed with paired-end Illumina Miseq 16S rDNA sequencing. The differences and changes on community level (alpha and beta diversity) and on the level of single agglomerated ribosomal sequence variants (aRSV) were calculated with analysis of covariance (ANCOVA) and likelihood ratio test (LRT). RESULTS: Interproximal and subgingival sites harbored predominately Fusobacterium and Prevotella species associated with periodontitis, whereas buccal/lingual sites harbored mainly Streptococcus and Veillonella species associated with periodontal health. Alpha and beta diversity did not change noticeably differently between both toothpaste groups (P > 0.05, ANCOVA). Furthermore, none of the aRSVs showed a noticeably different change between the tested toothpastes during periodontal therapy (Padj . > 0.05, LRT). CONCLUSION: The use of a toothpaste containing anti-adhesive HA did not induce statistically noticeably different changes on microbial composition compared to an antimicrobial and anti-adhesive AmF/SnF2 formulation.

Effects of stannous fluoride on eroded enamel permeability.

This study aimed to evaluate the effect in vitro of a single application of a stannous fluoride- (SnF2-) containing toothpaste on eroded enamel. Forty-eight teeth were subjected to three acid treatments: 15% hydrochloric acid for 120 s (HA group); 1% citric acid (pH=4) for 180 s (CA group); 37% phosphoric acid for 30 s (PA group). They were brushed with an electric toothbrush with pressure control and 1 g of SnF2 (1100 ppm) toothpaste for 2 min. Polyether replicas of buccal enamel surfaces were obtained at baseline, after acid exposure and after brushing, gold sputtered and inspected by SEM for fluid droplets presence. Hydrochloric and citric acid treatments increased enamel permeability while, on the contrary, phosphoric acid reduced enamel fluid release. SnF2 application of ameliorated acid induced permeability in citric and hydrochloric treated samples. Permeability in phosphoric treated enamel was unchanged after topical application of SnF2. Our data show specific acid-dependent effects on enamel permeability and demonstrate that SnF2 application can reverse acid-induced permeability.

Clinical effects of stannous fluoride dentifrice in reducing plaque microbial virulence III: Lipopolysaccharide and TLR2 reporter cell gene activation.

PURPOSE: This study expanded the analysis of subgingival dental plaques from previous research to include the evaluation of cohort, site and treatment effects on chemically measured endotoxin and activation of Toll-like receptor (TLR) based gene expression in two additional reporter cell lines: a TLR2 specific cell line and a THP-1 (multi TLR reporter) cell line. METHODS: Participants from high and low bleeding cohorts were sampled at baseline for both supra and subgingival dental plaque at both healthy as well as clinically diseased sites and then provided with intervention hygiene products including a stabilized SnF2 dentifrice and a new soft bristle manual toothbrush. Following 2 and 4 weeks of assigned dentifrice use, participants returned for a re-evaluation of gingival inflammation and bleeding and repeat samplings of dental plaque. Subgingival sampled plaques were chemically analyzed for endotoxin concentration using a Thermo Scientific Pierce LAL chromogenic endotoxin quantitation kit. Samples were also used for inoculation of two reporter cell assays (an HEK293 TLR2 reporter cell line and a THP-1 monocyte cell line). Reporter cell activation was analyzed via luminescence changes of secreted embryonic alkaline phosphatase. RESULTS: The endotoxin content of subgingival plaque could be measured directly with dye assays and plaque isolates activated gene expression in both TLR reporter cell lines. Higher disease cohorts and sites with gingival inflammation generally showed more endotoxins and higher levels of plaque virulence as compared to low disease cohorts or plaque sampled from clinically healthy sites. SnF2 dentifrice treatment was associated with broad scale reductions in endotoxin content and virulence potentiation properties of dental plaque samples collected subgingivally from patients. CLINICAL SIGNIFICANCE: These results collectively support the use of dye or various reporter cell lines in the characterization of plaque virulence in diseased populations and as a potential route for analysis in clinical evaluations of treatment interventions. Subgingival plaque 'detoxification' including effects on microbial pathogenicity as well as metabolic activity may be considered important mechanisms contributing to clinical benefits of SnF2 dentifrice.

Stannous Fluoride Effects on Gene Expression of Streptococcus mutans and Actinomyces viscosus.

A genome-wide transcriptional analysis was performed to elucidate the bacterial cellular response of Streptococcus mutans and Actinomyces viscosus to NaF and SnF2. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of SnF2 were predetermined before microarray study. Gene expression profiling microarray experiments were carried out in the absence (control) and presence (experimental) of 10 ppm and 100 ppm Sn2+ (in the form of SnF2) and fluoride controls for 10-min exposures (4 biological replicates/treatment). These Sn2+ levels and treatment time were chosen because they have been shown to slow bacterial growth of S. mutans (10 ppm) and A. viscosus (100 ppm) without affecting cell viability. All data generated by microarray experiments were analyzed with bioinformatics tools by applying the following criteria: 1) a q value should be ≤0.05, and 2) an absolute fold change in transcript level should be ≥1.5. Microarray results showed SnF2 significantly inhibited several genes encoding enzymes of the galactose pathway upon a 10-min exposure versus a negative control: lacA and lacB (A and B subunits of the galactose-6-P isomerase), lacC (tagatose-6-P kinase), lacD (tagatose-1,6-bP adolase), galK (galactokinase), galT (galactose-1-phosphate uridylyltransferase), and galE (UDP-glucose 4-epimerase). A gene fruK encoding fructose-1-phosphate kinase in the fructose pathway was also significantly inhibited. Several genes encoding fructose/mannose-specific enzyme IIABC components in the phosphotransferase system (PTS) were also downregulated, as was ldh encoding lactate dehydrogenase, a key enzyme involved in lactic acid synthesis. SnF2 downregulated the transcription of most key enzyme genes involved in the galactose pathway and also suppressed several key genes involved in the PTS, which transports sugars into the cell in the first step of glycolysis.

Do Abrasives Play a Role in Toothpaste Efficacy against Erosion/Abrasion?

Abrasives may counteract the efficacy of anti-erosion toothpastes either due to physical effects or due to interaction with active agents. This study aimed to investigate whether the amount of abrasives is a determinant for the efficacy of Sn2+-containing toothpastes with or without chitosan additive. Enamel samples were eroded (0.50 wt% citric acid, pH 2.5; 6 × 2 min/day) on a shaking desk - 30/min in experiment 1 (E1) and 35/min in experiments 2 (E2) and 3 (E3) - and immersed in toothpaste slurries (2 × 2 min). Half of the samples were additionally brushed (15 s, load 200 g) within the immersion time. The toothpastes contained 0, 5, 10, 15, and 20% silica. In E1 and E2 the active ingredients were F- (700 ppm as amine fluoride, 700 ppm as NaF) and Sn2+ (3,500 ppm as SnCl2); in E3 chitosan (0.5%) was additionally added. The placebo contained 20% silica. Tissue loss was determined profilometrically. In E1, slurries completely inhibited tissue loss; distinct surface deposits occurred. With brushing, tissue loss significantly increased up to an abrasive content of 10%, but decreased significantly with higher amounts; 20% silica revealed similar values as the abrasive-free formulation. In E2, all slurries inhibited tissue loss distinctly irrespective of the amounts of abrasives. With brushing, a similar trend as in E1 was observed but with much less efficacy. The chitosan-containing formulations in E3 were much more effective; similar results as in E1 were found. In conclusion, the amount of abrasives had no effect when toothpastes were applied as slurries, but played an important role with brushing.

Erosion protection benefits of stabilized SnF2 dentifrice versus an arginine-sodium monofluorophosphate dentifrice: results from in vitro and in situ clinical studies.

OBJECTIVES: The aim of these investigations was to assess the ability of two fluoride dentifrices to protect against the initiation and progression of dental erosion using a predictive in vitro erosion cycling model and a human in situ erosion prevention clinical trial for verification of effectiveness. MATERIALS AND METHODS: A stabilized stannous fluoride (SnF2) dentifrice (0.454 % SnF2 + 0.077 % sodium fluoride [NaF]; total F = 1450 ppm F) [dentifrice A] and a sodium monofluorophosphate [SMFP]/arginine dentifrice (1.1 % SMFP + 1.5 % arginine; total F = 1450 ppm F) [dentifrice B] were tested in a 5-day in vitro erosion cycling model and a 10-day randomized, controlled, double-blind, two-treatment, four-period crossover in situ clinical trial. In each study, human enamel specimens were exposed to repetitive product treatments using a standardized dilution of test products followed by erosive acid challenges in a systematic fashion. RESULTS: Both studies demonstrated statistically significant differences between the two products, with dentifrice A providing significantly better enamel protection in each study. In vitro, dentifrice A provided a 75.8 % benefit over dentifrice B (p < 0.05, ANOVA), while after 10 days in the in situ model, dentifrice A provided 93.9 % greater protection versus dentifrice B (p < 0.0001, general linear mixed model). CONCLUSION: These results support the superiority of stabilized SnF2 dentifrices for protecting human teeth against the initiation and progression of dental erosion. CLINICAL RELEVANCE: Stabilized SnF2 dentifrices may provide more significant benefits to consumers than conventional fluoride dentifrices.

Effect of mouthwashes on the composition and metabolic activity of oral biofilms grown in vitro.

OBJECTIVE: The aim of this study was to determine the effect of an oxygenating mouthwash compared to two other established mouthwash products on bacterial composition and metabolic activity of oral biofilms in vitro. MATERIAL AND METHODS: Twelve healthy subjects participated as donors. Plaque-saliva mixture inoculated biofilms were grown and treated with 3 different chemotherapeutic mouthwashes [amine fluoride/stannous fluoride (MD), oxygenating agent (AX), chlorhexidine 0.12 % (PA), and water (W)]. Effects of treatments were assessed on biofilm composition (16S rRNA gene amplicon sequencing), production of organic acids (formate, acetate, lactate, propionate, butyrate using capillary electrophoresis), and viability of the remaining biofilm (CFUs). RESULTS: Microbial profiles of biofilms clustered per inoculum donor and were dominated by the genera Veillonella, Streptococcus, and Prevotella. Microbial diversity was only reduced after PA treatment. Significant changes in composition occurred after treatment with AX, resulting in lower proportions of Veillonella and higher proportions of non-mutans streptococci. Production of all organic acids after PA and lactate after MD was significantly lower as compared to W. AX resulted in reduction of acetate, butyrate, and propionate and increase in lactate production (p < 0.05). Viable counts were significantly lower after PA and AX treatments compared to W, while no significant reduction was observed after MD. CONCLUSIONS: All studied mouthwashes affected the in vitro biofilms differently. The effects of the AX treatment were the most prominent which resulted in changes of the bacterial composition and metabolism. CLINICAL IMPLICATIONS: Awareness by the dental team that mouthwashes can change the bacterial composition and metabolism is important when advising its use.

Antibacterial Effects of Toothpastes Evaluated in an In Vitro Biofilm Model.

PURPOSE: To test the antibacterial effects of different toothpastes with the slurry method of toothpaste application in an in vitro oral biofilm model including relevant periodontal pathogens. MATERIALS AND METHODS: Four commercially available toothpastes, two containing sodium fluoride (NaF) at different concentrations (1450 and 2500 ppm), two NaF with either triclosan or stannous fluoride, and a control phosphate-buffered saline (PBS) were used. Multispecies biofilms containing 6 species of oral bacteria were grown on hydroxyapatite disks for 72 h and then exposed for 2 min to the toothpaste slurries or phosphate buffer saline (PBS) by immersion, under continuous agitation at 37°C. Biofilms were then analysed by means of real-time polymerase chain reaction (PCR), combined with propidium monoazide (PMA). Statistical evaluation was performed using ANOVA and Student's t-test, with Bonferroni correction for multiple comparisons. RESULTS: The toothpastes containing NaF and stannous fluoride demonstrated superior antimicrobial activity for A. actinomycetencomitans, P. gingivalis and F. nucleatum when compared to those containing NaF and triclosan, 1450 ppm NaF or 2500 ppm NaF in this multispecies biofilm model. CONCLUSION: The proposed model for the evaluation of toothpastes in the form of slurries detected significant differences in the antimicrobial effects among the tested NaF-containing toothpastes, with the stannous fluoride-based formulation achieving better results than the other formulations. The use of toothpaste as slurries and real-time PCR with PMA is an adequate method for comparing the in vitro antimicrobial effect of different toothpastes.

Re- and Demineralization Characteristics of Enamel Depending on Baseline Mineral Loss and Lesion Depth in situ.

OBJECTIVES: The aim of this double-blinded, randomized, cross-over in situ study was to evaluate the re- and demineralization characteristics of sound enamel as well as lowly and highly demineralized caries-like enamel lesions after the application of different fluoride compounds. METHODS: In each of three experimental legs of 4 weeks, 21 participants wore intraoral mandibular appliances containing 4 bovine enamel specimens (2 lowly and 2 highly demineralized). Each specimen included one sound enamel and either one lowly demineralized (7 days, pH 4.95) or one highly demineralized (21 days, pH 4.95) lesion, and was positioned 1 mm below the acrylic under a plastic mesh. The three randomly allocated treatments (application only) included the following dentifrices: (1) 1,100 ppm F as NaF, (2) 1,100 ppm F as SnF2 and (3) 0 ppm F (fluoride-free) as negative control. Differences in integrated mineral loss (x0394;x0394;Z) and lesion depth (x0394;LD) were calculated between values before and after the in situ period using transversal microradiography. RESULTS: Of the 21 participants, 6 did not complete the study and 2 were excluded due to protocol violation. Irrespectively of the treatment, higher baseline mineral loss and lesion depth led to a less pronounced change in mineral loss and lesion depth. Except for x0394;x0394;Z of the dentifrice with 0 ppm F, sound surfaces showed significantly higher x0394;x0394;Z and x0394;LD values compared with lowly and highly demineralized lesions (p < 0.05, t test). CONCLUSION: Re- and demineralization characteristics of enamel depended directly on baseline mineral loss and lesion depth. Treatment groups should therefore be well balanced with respect to baseline mineral loss and lesion depth.

In vitro Efficacy of Experimental Chitosan-Containing Solutions as Anti-Erosive Agents in Enamel.

The present study evaluated the effect of chitosans with different viscosities, dissolved in an AmF/SnCl2 solution, against erosion or erosion/abrasion. A total of 192 specimens were assigned to 2 × 6 groups (n = 16 specimens each): negative control, 4 chitosan solutions (groups Ch50, Ch500, Ch1000, and Ch2000, with viscosity of 50, 500, 1,000, or 2,000 mPas, respectively, 0.5% chitosan, 500 ppm F-, 800 ppm Sn2+, pH 4.4), and positive control (500 ppm F-, 800 ppm Sn2+, pH 4.3). One half of the groups was demineralized (experiment 1, E1; 10 days, 6 × 2 min/day, 0.5% citric acid, pH 2.8) and exposed to solutions (2 × 2 min/day); the other half was additionally brushed (15 s, 200 g) with non-fluoridated toothpaste before solution immersion (experiment 2, E2). Treatment effects were investigated by profilometry, energy-dispersive X-ray spectroscopy and scanning electron microscopy (SEM). In E1, all the chitosan-containing solutions reduced enamel loss by 77-80%, to the same extent as the positive control, except for Ch2000 (p ≤ 0.05), which completely inhibited tissue loss by the formation of precipitates. In E2, Ch50 and Ch500 showed best performance, with approximately 60% reduction of tissue loss compared to the negative control group (p ≤ 0.05 compared to other groups). SEM analysis showed differences between negative control and the other groups but only minor differences amongst the groups treated with active agents. In both E1 and E2, treatment with active agents resulted in surface enrichment of carbon and tin compared to negative control (p ≤ 0.001); brushing removed parts of carbon and tin (p ≤ 0.001). Chitosan shows different properties under erosive and erosive/abrasive conditions. Under erosive conditions high viscosity might be helpful, whereas lower viscosity seems to be more effective in cases of chemo-mechanical challenges.

Effects of Erosion Protocol Design on Erosion/Abrasion Study Outcome and on Active Agent (NaF and SnF2) Efficacy.

There is no standard for testing anti-erosive/anti-abrasive agents, making the assessment and comparison of study results difficult. Factors which are varied in study designs are amongst others the erosive medium regarding concentration and pH or movement type of acid. The present study therefore investigated the impact of these factors on dimension of tissue loss and on efficacy of active agents used as anti-erosive/anti-abrasive therapeutics. In 8 experiments, consisting of 8 groups each (n = 20 each), resulting in a total of 64 groups, enamel specimens were demineralised (10 days, 6 × 2 min/day) using different acids (1, 0.5 and 0.3% citric acid at native pH 2.3, 2.5 and 2.8, respectively, and 0.3% citric acid adjusted to pH 3.6) with two different movement types (jerky or smooth). Specimens were immersed (2 × 2 min/day) in slurries of 1,450 ppm F- toothpaste (NaF), 1,450 ppm F- and 3,436 ppm Sn2+ toothpaste (NaF/SnF2), 970 ppm F- and 3,000 ppm Sn2+ gel (SnF2) or placebo, or were additionally brushed during immersion (15 s, 200 g). All groups were in between stored in a mineral salt solution. Tissue loss was determined profilometrically. Movement type, pH and concentration of acid had a substantial impact on study outcome. The combination of jerky movement and concentrated acid masked, to some extent, differences between erosive and erosive-abrasive tissue loss. The acid at low concentration (0.3%), independent of pH, was too mild to produce any tissue loss. The model with the best ability to demonstrate effects of abrasive impacts and active agents used the 1% acid concentration combined with smooth acid movements.