The effect of silver diammine fluoride on In Vitro Enamel caries lesion remineralization and staining as a function of lesion baseline mineral distribution.

OBJECTIVES: to investigate whether baseline mineral distribution modulates the ability of silver diammine fluoride (SDF) to remineralize and stain enamel caries lesions. METHODS: This laboratory study followed a 3 [treatment: SDF/fluoride varnish (FV)/deionized water (DIW)] ×3 [lesion protocol: methylcellulose (MeC)/hydroxyethylcellulose (HEC)/Carbopol 907 (C907)] factorial design. Lesions were created in bovine enamel specimens (n = 20). Treatments were applied and lesions remineralized in artificial saliva. Digital transverse microradiography (TMR-D) was used to analyze lesions. Lesion color was monitored spectrophotometrically. The effects of lesion protocol and treatment on changes in lesion depth (ΔLD), mineral loss (ΔΔZ), maximum mineral density at the surface zone (ΔSZmax), and color changes related to remineralization (ΔL*remin) were analyzed using two-way ANOVA. RESULTS: The treatment×lesion protocol interaction was significant for ΔΔZ (p < 0.01) and ΔL*remin (p < 0.01), however not for ΔLD (p = 0.23) or ΔSZmax (p = 0.91). There were no differences in ΔΔZ between treatments in HEC and C907 lesions. However, DIW resulted in more remineralization than both SDF (p < 0.01) and FV (p = 0.01) in MeC lesions. Considering changes from lesion baseline after remineralization in MeC lesions, SDF treatment resulted in the highest mineral gain in the surface zone. However, DIW revealed the highest mineral gain after remineralization in the lesion body. SDF stained lesions with the intensity increasing after remineralization in C907 lesions, whereas staining decreased in MeC and HEC lesions. CONCLUSION: High fluoride treatments can interfere with continuous remineralization of caries lesions due to partial arrest. Baseline lesion mineral distribution affects SDF's ability to enhance remineralization and the staining caused by SDF. CLINICAL SIGNIFICANCE: SDF is being used to arrest active caries lesions extending into dentin and to treat dentin hypersensitivity. This study shed light on SDF's effect on an isolated process in dental caries only, remineralization. It achieved this by examining enamel caries lesions with differing mineral distributions and assessing their staining properties.

Characteristics of tin-containing fluoride toothpastes related to erosive tooth wear protection.

OBJECTIVES: To assess the effect of different tin-containing toothpastes on the control of erosive tooth wear in enamel and dentin. METHODS: Enamel and dentin slabs were randomly distributed into 7 experimental groups (n = 10/substrate): C-: negative control (Artificial saliva); AmF (regular fluoridated toothpaste without tin); Sn-1 (SnF2/NaF); Sn-2 (SnF2/NaF/SnCl2); Sn-3 (SnCl2/NaF); Sn-4 (SnF2/SnCl2); Sn-5 (SnCl2/AmF/NaF/chitosan). Specimens were submitted to 5-day erosion-abrasion cycling. Surface loss (SL) was determined with an optical profilometer. Tin deposition on the tooth surfaces and some characteristics of the toothpastes (pH, potentially available F-, %weight of solid particles, and RDA) were also assessed. Data were statistically analyzed (α = 0.05). RESULTS: For enamel, the Sn-2 presented the lowest SL, not differing significantly from AmF, C+, and Sn-3. The SL of these groups was significantly lower than the C-, except for Sn-3. Sn-1 and Sn-4 were also not significantly different from C-. For dentin, C- significantly showed the highest SL values, whilst, Sn-1 presented the lowest SL, not differing significantly from AmF, Sn-2, C+, and Sn-3. There was a significant positive association between enamel SL and the pH and tin deposition. Dentin SL was significantly negatively associated with the %weight of solid particles and RDA. CONCLUSIONS: Most of the tin-toothpastes were able to exhibit some protection against ETW. In this process, the toothpastes characteristics play a role, as lower enamel SL was significantly associated with lower pH values and tin deposition; and lower dentin SL was associated with higher %weight of solid particles and RDA of the toothpastes. CLINICAL SIGNIFICANCE: Tin-containing toothpastes can be used for erosive tooth wear protection, but our study showed that their effect depends on the pH, amount of tin deposition, % weight of solid particles and RDA of the toohpastes.

Objective assessment of simulated non-carious cervical lesion by tridimensional digital scanning - An in vitro study.

OBJECTIVE: Clinical assessment of progression of non-carious cervical lesions (NCCLs) is currently based on subjective methods. We hypothesize that 3D digital intraoral scanners (IOS) can provide quantitative outcomes for objective measuring and monitoring of NCCLs. This in vitro study verified the reliability of IOS to monitor dental structure loss of simulated NCCLs, in comparison to a reference bench-top 3D optical profilometer (BTS). METHODS: NCCLs of different severity levels (early, moderate, or severe, n = 150) were simulated in a preceding study. Impressions of the lesions were taken and scanned with both BTS (ProScan; Scantron) and IOS (TRIOS4; 3Shape). Generated 3D images were analyzed for volumetric tooth loss (mm3) by superimposition followed by subtraction analysis. ProForm (Scantron) and WearCompare (Leeds Digital Dentistry) software were used in association to BTS and IOS, respectively. Agreement was assessed using intraclass correlation coefficient (ICC, alpha=0.05) and Bland-Altman plots. RESULTS: ICC (confidence interval at 95 %) between IOS and BTS for all data combined was 0.962 (0.942-0.973), showing excellent reliability. Subset analyses showed that NCCLs with lower volume loss (early- and moderate-stage lesions) resulted in moderate ICCs, whereas severe lesions showed excellent ICC. Bland-Altman plots demonstrated general good agreement, with narrower limits for early stage-lesions. CONCLUSION: IOS data demonstrated good agreement to BTS, when measuring tooth structure loss in simulated NCCLs, with particularly higher ICC in more severe lesions. Considering the accessibility and ease-of-use, IOS showed to be a good alternative for the objective assessment of NCCLs in vitro. CLINICAL SIGNIFICANCE: 3D intraoral scanners' accessibility and objectivity make them potentially valuable tools for assessing and monitoring NCCLs.

Effect of Three Chemical Agents on Stain Removal from Dentin Caries Lesions Treated with Silver Diamine Fluoride.

INTRODUCTION: This in vitro study aimed to evaluate the effect of three different chemical agents on stain removal and mineral uptake of artificial dentin caries (ADC) lesions treated with silver diamine fluoride (SDF). METHODS: Baseline L*a*b* values were determined in polished human permanent dentin blocks, and ADC lesions were induced with an acid gel for 1 week. Samples were assigned to four groups; in three groups, half of each sample received SDF (30% SDF for 3 min), while the other half received SDF followed by a bleaching treatment protocol (garlic extract, bentonite, or 35% hydrogen peroxide). The fourth group had one SDF-treated half and one half without SDF. Color changes (ΔE) were measured by spectrometry, and transversal microradiography was used to quantify integrated mineral loss (ΔZ) 24 h after treatment (SDF or SDF + bleaching). A two-way mixed ANOVA was applied to thirty percent. RESULTS: SDF application increased mineral uptake by ADC (p = 0.001). The type of chemical agent evaluated (p < 0.0001), time (p = 0.01), and their interaction (p < 0.0001) bleached the ADC treated with SDF. However, 35% hydrogen peroxide was the only compound with a bleaching effect (p < 0.001), without returning to baseline color. None of the compounds altered the mineral uptake effect of SDF (p = 0.30). CONCLUSION: This in vitro study showed mineral uptake effect in ACD within 24 h after SDF application and the ability of hydrogen peroxide to partially remove (reduction of 24%) the staining caused by SDF without affecting its mineral uptake effect.

Impact of toothbrush head configuration and dentifrice abrasivity on non-carious cervical lesions in-vitro.

OBJECTIVE: To investigate the effect of toothbrush head configuration and dentifrice slurry abrasivity on the development of simulated non-carious cervical lesions (NCCLs) in vitro. METHODS: Extracted premolars were randomly allocated into 15 groups (n = 16) generated by the association between toothbrush head configuration (flat-trimmed, rippled, cross-angled/multilevel/rubbers added, cross-angled/multilevel/flex head, feathered) and dentifrice slurry abrasivity (low/medium/high). Teeth were mounted on acrylic blocks and had their roots partially covered with acrylic resin, leaving 2-mm root surfaces exposed. Toothbrushing was performed for 35,000 and 65,000 double-strokes. Specimens were analyzed using non-contact profilometry for dental volume loss (mm3) and lesion morphology. Data was analyzed using ANOVA with pairwise comparisons and Kruskal-Wallis tests. RESULTS: The two-way interaction between toothbrush head configuration and slurry abrasivity was significant (p = 0.02). At 35,000 strokes, the flat-trimmed and cross-angled/multilevel/rubbers added toothbrushes caused the highest loss, when associated to the high-abrasive slurry (p<0.05); whereas cross-angled/multilevel/flex head showed the least loss, when associated to the low-abrasive (p<0.05). At 65,000, more dental loss was observed for all toothbrushes when associated to the high-abrasive slurry, with flat-trimmed causing the highest loss (p < 0.05). Lower dental loss rates were observed for cross-angled/multilevel/flex head associated to the low-abrasive slurry when compared to the other toothbrushes (p < 0.05), except to feathered (p = 0.14) and rippled (p = 0.08). Flat lesions (mean internal angle ± standard-deviation: 146.2°± 16.8) were mainly associated with low-abrasive slurry, while wedge-shaped lesions (85.8°± 18.8) were more frequent with medium- and high-abrasive slurries. CONCLUSION: The development, progression and morphology of simulated NCCLs were modulated by both toothbrush head configuration and dentifrice abrasivity. CLINICAL SIGNIFICANCE: Dental professionals should consider both the type of toothbrush and dentifrice abrasivity in the management plan of patients at risk of developing NCCLs.

Novel resin-based material containing ß-tricalcium phosphate nanoparticles for the reduction of dentin permeability.

OBJECTIVES: To synthesize and characterize a novel dentin adhesive containing Beta-Tricalcium Phosphate (ß-TCP) nanoparticles and test its ability to reduce dentin permeability (dP). METHODS: Experimental adhesives were prepared by mixing Bis-GMA, TEGDMA, HEMA (50/25/25 wt.%), photo-initiators, and inhibitors. The following groups were tested: Experimental adhesives without ß-TCP (Exp.); with 10 wt.% ß-TCP (Exp.10 wt.% ß-TCP); with 15 wt.% ß-TCP (Exp.15 wt.% ß-TCP), Scotchbond Multi-Purpose (SBMP) and Clearfil SE Protect Bond (CFPB). Degree of conversion (DC%, 10 and 20 s); Flexural Strength (FS), Knoop Hardness (KHN), and Cell Viability (OD%) tests were performed. dP was evaluated by hydraulic conductance, using human dentin disks (n=12), at three-time intervals: post-EDTA (T0); post-treatment (T1); and post-erosion/abrasion cycling (T2). Data were statistically analyzed (α=0.05). RESULTS: For all groups, exposure time for 20 s presented a higher DC% than for 10 s. For FS, filled adhesives did not differ from unfilled and from CFPB. Experimental adhesives did not differ among them and showed lower KHN than the commercial products. Cell viability did not differ among adhesives, except Exp. 15 wt.%, which showed lower OD% than Exp., Exp. 10% and, CFPB. For dP, only Exp.10 and 15 wt.% ß-TCP did not present difference between the times T1 and T2. After cycling, Exp.10 wt.% ß-TCP presented lower permeability than Exp. and CFPB. CONCLUSIONS: The incorporation of 10 wt.% ß-TCP nanoparticles into the resin-based dental material did not affect its mechanical properties and biocompatibility, and promoted the greatest reduction in dentin permeability, sustaining this effect under erosive/abrasive challenges. CLINICAL SIGNIFICANCE: A novel resin-based dental material containing ß-TCP nanoparticles was able to reduce dentin permeability, maintaining its efficacy after erosive/abrasive challenges. The synthesized material did not affect dental pulp cell viability and might be promising for other conditions that require dental remineralization, such as tooth wear and dental caries.

Fluorotic Enamel Susceptibility to Dental Erosion and Fluoride Treatment.

The purpose of this in vitro study was to test the hypothesis that fluoride treatment can prevent dental erosion on fluorotic enamel of different severities. It followed a 3×2 factorial design, considering a) fluorosis severity: sound (TF0, Thylstrup-Fejerskov Index), mild (TF1-2), moderate (TF3-4); and b) fluoride treatment: 0 (negative control) and 1150ppmF. Human molars with the three fluorosis severities (n=16, each) were selected and randomly assigned to the two fluoride treatments (n=8). Enamel blocks (4×4mm) were prepared from each tooth and subjected to a dental erosion cycling model, for 10 days. The daily cycling protocol consisted of erosive challenges (1% citric acid, pH 2.4), interspersed by periods of immersion in artificial saliva, and three 2-minute treatments with either 0 or 1150ppm F. The enamel volume loss (mm3) was calculated by subtracting values obtained by microtomography before and after cycling. Two-Way ANOVA showed no significant interaction between fluorosis severity and fluoride treatment (p=0.691), and no significant effect for either fluorosis severity (TF0 mean±standard-deviation: 13.5(10-2±0.42(10-2, TF1-2: 1.50(10-2±0.52(10-2, TF3-4: 1.24(10-2±0.52(10-2, p=0.416) or treatment (0ppmF: 1.49(10-2±0.53(10-2; 1150ppmF: 1.21(10-2±0.42(10-2; p=0.093), when evaluated independently. Considering the limitations of this in vitro study, the presence and severity of fluorosis in enamel do not appear to affect its susceptibility to dental erosion. Fluoride treatment was not effective in preventing the development of dental erosion in both sound and fluorotic enamel substrates under our experimental conditions.

Longitudinal assessment of dental erosion-abrasion by cross-polarization optical coherence tomography in vitro.

This study tested a novel in vitro dental erosion-abrasion model and the performance of cross-polarization optical coherence tomography (CP-OCT) in longitudinally monitoring the simulated lesions. Thirty human enamel specimens were prepared and randomized to receive three dental erosion-abrasion (EA) protocols: severe (s-EA, lemon juice/pH:2.5/4.25%w/v citric acid), moderate (m-EA, grapefruit juice/pH:3.5/1.03%w/v citric acid) and no-EA (water, control). EA challenge was performed by exposing the specimens to acidic solutions 4x/day and to brushing 2x/day with 1:3 fluoridated toothpaste slurry, for 14 days. Enamel thickness measurements were obtained using CP-OCT at baseline (D0), 7 (D7) and 14 days (D14) and micro-computed tomography (micro-CT) at D14. Enamel surface loss was measured with both CP-OCT and optical profilometry at D0, D7 and D14. Data was analyzed with repeated-measures ANOVA and Pearson's correlation (r) (α = 0.05). CP-OCT enamel thickness decreased over time in the s-EA group (D0 >D7 > D14, p < 0.001) and m-EA group (D0 > D14, p = 0.019) but did not change in the no-EA group (p = 0.30). Overall, CP-OCT and micro-CT results at D14 correlated moderately (r = 0.73). CP-OCT surface loss was highest for s-EA (p <0.001) but did not differ between moderate and no-EA (p = 0.25). Enamel surface loss with profilometry increased with severity (no-EA>m-EA>s-EA, p < 0.001). D14 surface loss was higher than D7 for both methods except for the no-EA group with profilometry. CP-OCT and profilometry had moderate overall correlation (r = 0.70). Our results revealed that the currently proposed in vitro dental erosion-abrasion model is valid and could simulate lesions of different severities over time. CP-OCT was a suitable method for monitoring the EA lesions.

Exploratory Analysis of Objective Outcome Measures for the Clinical Assessment of Erosive Tooth Wear.

This study proposed using enamel surface texture and thickness for the objective detection and monitoring of erosive tooth wear (ETW), comparing them to the standard subjective Basic Erosive Wear Evaluation (BEWE). Thirty-two subjects (n = 597 teeth) were enrolled in this longitudinal observational clinical study. Enamel thickness (by cross-polarization optical coherence tomography, CP-OCT) and 3D dental microwear parameters, i.e., area-scale fractal complexity (Asfc), anisotropy (Str), and roughness (Sa) (by white-light scanning confocal profilometry), were obtained from buccal surfaces. Buccal, occlusal, and lingual surfaces were scored for BEWE and the maximum score per tooth (BEWEMax) was determined at baseline and 12 months (M12). Data outcome relationships were evaluated (alpha = 0.05). Enamel thickness decreased (p < 0.001), BEWE scores, Sa, and Str increased (p < 0.001), while Asfc did not change at M12. Baseline BEWEBuccal correlated strongly with BEWEMax (r = 0.86, p < 0.001) and moderately with BEWELingual (r = 0.42, p < 0.001), but not with enamel thickness (r = 0.03, p = 0.43). Change (Δ) in surface texture outcomes correlated poorly but significantly with ΔBEWEBuccal (r = -0.15-0.16, p < 0.001) and did not correlate with Δenamel thickness (r = 0.02-0.09, p > 0.06). Teeth with BEWE progression revealed a greater increase in ΔSa and ΔStr. These findings suggest that enamel surface roughness can potentially determine ETW severity, and CP-OCT may be relevant for clinically monitoring enamel thickness.

Susceptibility of fluorotic enamel to dental erosion-abrasion.

Dental hard tissue conditions can be of pre- or post-eruptive nature, such as enamel fluorosis and erosive tooth wear (ETW), respectively. Dental enamel fluorosis is caused by the chronic and excessive intake of fluoride during enamel development, leading to increased fluoride concentration and increased porosity. ETW has become a common clinical condition and often impairs dental function and aesthetics. This in vitro study tested the hypothesis that fluorotic enamel presents different susceptibility to dental erosion-abrasion. It consisted of a 3×3×2 factorial design, considering a) fluorosis severity: sound (TF0), mild (TF1-2), moderate (TF3-4); b) abrasive challenge: low, medium, and high; and c) erosive challenge: yes or no. A total of 144 human teeth were selected according to the three fluorosis severity levels (n=48), and subdivided into six groups (n = 8) generated by the association of the different erosive and abrasive challenges. Enamel blocks (4×4 mm) were prepared from each tooth and their natural enamel surfaces subjected to an erosion-abrasion cycling model. After cycling, the depth of the lesions in enamel was assessed by profilometry. ANOVA showed that the three-way and two-way interactions among the factors were not significant (p > 0.20). Enamel fluorosis level (p=0.638) and abrasion level (p = 0.390) had no significant effect on lesion depth. Acid exposure caused significantly more enamel surface loss than water (p < 0.001). Considering the limitations of this in vitro study, fluorosis did not affect the susceptibility of enamel to dental erosion-abrasion.

Relationships between Dental Topography, Gross Wear, and Bang and Ramm/Liversidge and Molleson Age Estimates for a Sample of Human Premolar Teeth.

Objectives: Molar crown wear is often used in bioarchaeological research as a proxy for age at death. However, a small number of researchers have used premolars or compared the application of different methods of relative age estimation. Material and methods: Using a sample of 197 previously extracted maxillary first premolars from US dental patients, we considered three protocols for estimating age: the Bang and Ramm/Liversidge and Molleson (BRLM) age estimate method, occlusal topographic analysis, and the Smith system of macrowear scoring. A previous study utilizing the Bang and Ramm method yielded an age estimate range of 9.4 to 10.8 years for the sample. Results: Our analyses showed no associations between occlusal topography parameters (occlusal slope, relief, or faceting) and BRLM age estimates, but some concordance was found between Smith scoring and BRLM ages estimates and between Smith scoring and occlusal topography parameters. Conclusion: The results of the present study suggest that relationships between gross tooth wear, tooth shape, and dental age estimates are complex, and available methods should be considered together to gain a more comprehensive understanding of how teeth change their shape with wear throughout the lifecourse.

Effect of a modified adhesive system with encapsulated arginine and calcium carbonate on dentin permeability.

To modify an adhesive system with halloysite clay nanotubes (HNTs) containing arginine and calcium carbonate and to evaluate their cytocompatibility, viscosity and efficacy in reducing dentin permeability. HNTs containing arginine and calcium carbonate were incorporated into the primer and adhesive of a three-step adhesive system (SBMP), and their viscosity was measured. Discs (n = 4/group) were prepared: SBMP (control), HNT-PR (modified primer), HNT-ADH (modified adhesive) and HNT-PR + ADH (modified primer and adhesive) were evaluated regarding cell death and viability. Dentin discs were prepared and randomly assigned into the following treatments (n = 10): NC (no treatment), SBMP, HNT-PR, HNT-ADH, HNT-PR + ADH and COL (Colgate® Sensitive Pro-relief™ prophylaxis paste). After, they were submitted to an erosive-abrasive cycling. Dentin permeability (hydraulic conductance) was evaluated at baseline, 24 h after treatment and after cycling. Both the modified primer and adhesive showed significantly higher viscosity than their controls. Group HNT-PR resulted in significantly higher cytotoxicity when compared to SBMP and HNT-PR + ADH groups. Group HNT-ADH resulted in the highest cell viability compared to all other groups. All groups showed significantly lower dentin permeability when compared to the NC group. Post-cycling, SBMP and HNT-ADH groups showed significantly lower permeability when compared to COL group. The addition of encapsulated arginine and calcium carbonate did not affect the cytocompatibility of the materials nor their ability to reduce dentin permeability.

Objective characterization of dental occlusal and fissure morphologies: Method development and exploratory analysis.

OBJECTIVE: To develop clinically applicable methods to characterize occlusal topography and assess possible associations between morphology and caries incidence and development. DESIGN: In this retrospective clinical study, we evaluated caries presence and severity pre- and post-orthodontic treatment for first molars of 147 patients (384 teeth). These teeth were previously scanned using a clinical intraoral scanner, and the obtained digital elevation models were used to 1) analyze the 3D occlusal surface parameters (n = 384) and 2) quantitatively characterize the mandibular molars' (n = 166) fissure patterns using three novel methods. Pearson correlation coefficients were calculated to evaluate the associations among the measurements, and presence/severity of caries pre- and post-treatment were assessed using generalized linear mixed-effects models. RESULTS: Robust quantitative fissure characterizations were developed, and reliable occlusal surface parameters were obtained. In the studied population, none of the parametric measurements (Slope: p = 0.62 for presence, p = 0.96 for severity; Relief Index (RFI): p = 0.36, p = 0.84; Orientation Patch Count rotated (OPCr): p = 0.48, p = 0.13; Dirichlet Normal Energy (DNE): p = 0.91, p = 0.15) or the fissure morphological measurements (Mesial Angle: p = 0.43; Distal Angle: p = 0.86; Average Angle: p = 0.52; Area Difference: p = 0.83; Percent Fissure: p = 0.68) were found to be significantly associated with caries status or severity. CONCLUSION: Despite the lack of correlation in the limited studied sample, the tools developed to characterize occlusal surface topography and fissure morphology have the potential to be used in more comprehensive clinical evaluations.

New Theory Illustrates the Formation of Enamel CUP-Shaped Lesions on Occlusal Surfaces.

We propose a new theory for enamel cupping lesions formation. At early stages, naturally formed cupping lesions showed increased porosity at two structural prismatic traits: the central cone extending into the enamel-dentine junction and the type-I Hunther-Schreger bands (HSB), suggesting them to be the main drivers for cupping lesion formation and development. In addition, these lesions were circumscribed by type-II HSBs, which present lower surface porosity and higher resistance to wear. This theory was verified in in vitro observations, where both the central cone and the type-I HSB of cuspal enamel showed higher susceptibility to wear, potentially elucidating the mechanisms involved on cupping lesion formation.

Fluorotic Enamel Susceptibility to Dental Erosion and Fluoride Treatment

Abstract The purpose of this in vitro study was to test the hypothesis that fluoride treatment can prevent dental erosion on fluorotic enamel of different severities. It followed a 3×2 factorial design, considering a) fluorosis severity: sound (TF0, Thylstrup-Fejerskov Index), mild (TF1-2), moderate (TF3-4); and b) fluoride treatment: 0 (negative control) and 1150ppmF. Human molars with the three fluorosis severities (n=16, each) were selected and randomly assigned to the two fluoride treatments (n=8). Enamel blocks (4×4mm) were prepared from each tooth and subjected to a dental erosion cycling model, for 10 days. The daily cycling protocol consisted of erosive challenges (1% citric acid, pH 2.4), interspersed by periods of immersion in artificial saliva, and three 2-minute treatments with either 0 or 1150ppm F. The enamel volume loss (mm3) was calculated by subtracting values obtained by microtomography before and after cycling. Two-Way ANOVA showed no significant interaction between fluorosis severity and fluoride treatment (p=0.691), and no significant effect for either fluorosis severity (TF0 mean±standard-deviation: 13.5(10-2±0.42(10-2, TF1-2: 1.50(10-2±0.52(10-2, TF3-4: 1.24(10-2±0.52(10-2, p=0.416) or treatment (0ppmF: 1.49(10-2±0.53(10-2; 1150ppmF: 1.21(10-2±0.42(10-2; p=0.093), when evaluated independently. Considering the limitations of this in vitro study, the presence and severity of fluorosis in enamel do not appear to affect its susceptibility to dental erosion. Fluoride treatment was not effective in preventing the development of dental erosion in both sound and fluorotic enamel substrates under our experimental conditions.

Resumo Este estudo in vitro foi testou a hipótese de que o tratamento com flúor pode prevenir a erosão dentária no esmalte fluorótico de diferentes severidades. O objetivo deste estudo foi: investigar o efeito protetor dos fluoretos contra a erosão e abrasão simuladas no esmalte fluorótico. Seguiu um desenho fatorial 3×2, considerando a) severidade da fluorose em 3 níveis: hígido (TF0, Índice Thylstrup-Fejerskov), suave (TF1-2), moderada (TF3-4); b) tratamento com flúor: 0 (controle negativo) e 1150ppmF. Molares humanos com as três severidades de fluorose (n=16, cada) foram selecionados e distribuídos aleatoriamente para os dois tratamentos com flúor (n=8). Blocos de esmalte (4×4mm) foram preparados a partir de cada dente e submetidos a um modelo de ciclo de erosão dentária, por 10 dias. O protocolo de ciclagem diária consistiu em seis desafios erosivos de 5 minutos (1% de ácido cítrico, pH 2,4), intercalados por seis períodos de imersão em saliva artificial e três tratamentos de 2 minutos com 0 ou 1150ppmF. O volume do esmalte perdido foi calculado subtraindo o perfil superficial 3D obtido por microtomografia antes e depois da ciclagem. A ANOVA de dois fatores não mostrou interação significativa entre a severidade da fluorose e o tratamento com flúor (p = 0,691) e nenhum efeito significativo para a severidade da fluorose (TF0 média+/desvio padrão: 13,5(10-2±0,42(10-2, TF1-2: 1,50(10-2±0,52(10-2, TF3-4: 1,24(10-2±0,52(10-2, p=0,416) ou tratamento (0: 1,49(10-2±0,53(10-2; 1150ppmF: 1,21(10-2±0,42(10-2, p=0,093), quando avaliados independentemente. Considerando as limitações deste estudo in vitro, a presença e severidade da fluorose no esmalte não parece afetar sua suscetibilidade à erosão dentária. O tratamento com flúor não foi eficaz na prevenção do desenvolvimento da erosão dentária em esmalte hígido e fluorótico, sob as condições experimentais utilizadas.

Longitudinal assessment of dental erosion-abrasion by cross-polarization optical coherence tomography in vitro

Abstract: This study tested a novel in vitro dental erosion-abrasion model and the performance of cross-polarization optical coherence tomography (CP-OCT) in longitudinally monitoring the simulated lesions. Thirty human enamel specimens were prepared and randomized to receive three dental erosion-abrasion (EA) protocols: severe (s-EA, lemon juice/pH:2.5/4.25%w/v citric acid), moderate (m-EA, grapefruit juice/pH:3.5/1.03%w/v citric acid) and no-EA (water, control). EA challenge was performed by exposing the specimens to acidic solutions 4x/day and to brushing 2x/day with 1:3 fluoridated toothpaste slurry, for 14 days. Enamel thickness measurements were obtained using CP-OCT at baseline (D0), 7 (D7) and 14 days (D14) and micro-computed tomography (micro-CT) at D14. Enamel surface loss was measured with both CP-OCT and optical profilometry at D0, D7 and D14. Data was analyzed with repeated-measures ANOVA and Pearson's correlation (r) (α = 0.05). CP-OCT enamel thickness decreased over time in the s-EA group (D0 >D7 > D14, p < 0.001) and m-EA group (D0 > D14, p = 0.019) but did not change in the no-EA group (p = 0.30). Overall, CP-OCT and micro-CT results at D14 correlated moderately (r = 0.73). CP-OCT surface loss was highest for s-EA (p <0.001) but did not differ between moderate and no-EA (p = 0.25). Enamel surface loss with profilometry increased with severity (no-EA>m-EA>s-EA, p < 0.001). D14 surface loss was higher than D7 for both methods except for the no-EA group with profilometry. CP-OCT and profilometry had moderate overall correlation (r = 0.70). Our results revealed that the currently proposed in vitro dental erosion-abrasion model is valid and could simulate lesions of different severities over time. CP-OCT was a suitable method for monitoring the EA lesions.

Susceptibility of fluorotic enamel to dental erosion-abrasion

Abstract Dental hard tissue conditions can be of pre- or post-eruptive nature, such as enamel fluorosis and erosive tooth wear (ETW), respectively. Dental enamel fluorosis is caused by the chronic and excessive intake of fluoride during enamel development, leading to increased fluoride concentration and increased porosity. ETW has become a common clinical condition and often impairs dental function and aesthetics. This in vitro study tested the hypothesis that fluorotic enamel presents different susceptibility to dental erosion-abrasion. It consisted of a 3×3×2 factorial design, considering a) fluorosis severity: sound (TF0), mild (TF1-2), moderate (TF3-4); b) abrasive challenge: low, medium, and high; and c) erosive challenge: yes or no. A total of 144 human teeth were selected according to the three fluorosis severity levels (n=48), and subdivided into six groups (n = 8) generated by the association of the different erosive and abrasive challenges. Enamel blocks (4×4 mm) were prepared from each tooth and their natural enamel surfaces subjected to an erosion-abrasion cycling model. After cycling, the depth of the lesions in enamel was assessed by profilometry. ANOVA showed that the three-way and two-way interactions among the factors were not significant (p > 0.20). Enamel fluorosis level (p=0.638) and abrasion level (p = 0.390) had no significant effect on lesion depth. Acid exposure caused significantly more enamel surface loss than water (p < 0.001). Considering the limitations of this in vitro study, fluorosis did not affect the susceptibility of enamel to dental erosion-abrasion.

Interkingdom assemblages in human saliva display group-level surface mobility and disease-promoting emergent functions.

Fungi and bacteria often engage in complex interactions, such as the formation of multicellular biofilms within the human body. Knowledge about how interkingdom biofilms initiate and coalesce into higher-level communities and which functions the different species carry out during biofilm formation remain limited. We found native-state assemblages of Candida albicans (fungi) and Streptococcus mutans (bacteria) with highly structured arrangement in saliva from diseased patients with childhood tooth decay. Further analyses revealed that bacterial clusters are attached within a network of fungal yeasts, hyphae, and exopolysaccharides, which bind to surfaces as a preassembled cell group. The interkingdom assemblages exhibit emergent functions, including enhanced surface colonization and growth rate, stronger tolerance to antimicrobials, and improved shear resistance, compared to either species alone. Notably, we discovered that the interkingdom assemblages display a unique form of migratory spatial mobility that enables fast spreading of biofilms across surfaces and causes enhanced, more extensive tooth decay. Using mutants, selective inactivation of species, and selective matrix removal, we demonstrate that the enhanced stress resistance and surface mobility arise from the exopolymeric matrix and require the presence of both species in the assemblage. The mobility is directed by fungal filamentation as hyphae extend and contact the surface, lifting the assemblage with a "forward-leaping motion." Bacterial cell clusters can "hitchhike" on this mobile unit while continuously growing, to spread across the surface three-dimensionally and merge with other assemblages, promoting community expansion. Together, our results reveal an interkingdom assemblage in human saliva that behaves like a supraorganism, with disease-causing emergent functionalities that cannot be achieved without coassembly.

Antagonistic interactions by a high H2 O2 -producing commensal streptococcus modulate caries development by Streptococcus mutans.

Dental caries (tooth-decay) is caused by biofilms harboring polymicrobial communities on teeth that leads to the onset of localized areas of enamel demineralization. Streptococcus mutans has been clinically associated with severe caries in childhood. Although commensal bacteria can combat S. mutans using self-generated antimicrobials such as hydrogen peroxide (H2 O2 ), constant sugar-rich diet consumption disrupts microbial homeostasis shifting toward cariogenic community. Recently, Streptococcus oralis subsp. tigurinus strain J22, an oral isolate, was identified as a uniquely potent H2 O2 producer. Here, we assess whether a high H2 O2 -producing commensal streptococcus can modulate the spatial organization and virulence of S. mutans within biofilms. Using an experimental biofilm model, we find that the presence of S. oralis J22 can effectively inhibit the clustering, accumulation, and spatial organization of S. mutans on ex vivo human tooth surface, resulting in significant reduction of enamel demineralization. Notably, the generation of H2 O2 via pyruvate oxidase (SpxB) from S. oralis J22 is not repressed by sugars (a common repressor in other mitis group streptococci), resulting in enhanced inhibition of S. mutans growth (vs. Streptococcus gordonii). We further investigate its impact on biofilm virulence using an in vivo rodent caries model under sugar-rich diet. Coinfection of S. mutans with S. oralis results in reduced caries development compared to either species infected alone, whereas coinfection with S. gordonii has negligible effects, suggesting that the presence of an efficient, high H2 O2 -producer can disrupt S. mutans virulence. This work demonstrates that oral isolates with unusual high H2 O2 production may be capable of modulating biofilm cariogenicity in vivo. The findings also highlight the importance of bacterial antagonistic interactions within polymicrobial communities in health and in disease-causing state.

Non-Biological and Biological Risk Indicators for Tooth Wear Outcomes in Adults.

This study was undertaken to estimate the prevalence and severity of tooth wear (TW), as well as to assess non-biological and biological risk indicators for TW outcomes in adults. A cross-sectional study, adhering to the reporting STROBE guidelines, was conducted and had a convenience sample of adults ≥18 years of age who had at least one bilateral posterior molar contact. A total of 570 participants, seeking consultation at the University Hospital in Brussels, Belgium, were allocated to three age groups: 18-34 years (n = 232), 35-54 years (n = 256), and ≥55 years old (n = 79). Participants answered a self-applied questionnaire regarding sociodemographics, oral hygiene, and lifestyle behavior. The questionnaire was tested-retested (intraclass correlation coefficient = 0.71). Inter-examiner reliability for clinical examinations was k = 0.76-0.80. The prevalence of TW was 75% (95% CI: 71.7-78.9). Out of these patients, 42% (95% CI: 38.0-46.3) had as the primary etiological factor the process of dental erosion, while 22% (95% CI: 18.9-25.9) and 11% (95% CI: 8.6-13.9) had the processes of dental attrition and dental abrasion, respectively. The severity of TW according to BEWE highest score was mild in 31%, moderate in 28%, and severe in 17% of participants. The hierarchical logistic regression model for the association between risk indicators and TW irrespective of the etiology was significant for age (>35-54 years: OR = 2.35 and ≥55 years: OR = 3.89; p < 0.001), gender (>male: OR = 2.03; p < 0.001), toothpaste (>sensitive teeth: OR = 2.34; p = 0.005), occlusal splint (>yes: OR = 1.62; p = 0.03), and acidic beverages (≥once per day: OR = 1.62; p = 0.044). Consumption of acidic beverages was not associated with TW having as the primary etiological factor the process of dental attrition or dental abrasion, while it was significantly associated with the process of dental erosion (>once per week: OR = 1.69; p = 0.043 and ≥once per day: OR = 1.73; p = 0.016). Medical conditions were equally associated with the latter (OR = 3.11; p < 0.001). These findings could contribute to improving the effectiveness and sustainability of awareness in contemporary adult populations. In conclusion, the prevalence and severity of TW in adults were substantial. Medical conditions and consumption of acidic beverages were risk indicators for TW having as the primary etiological factor the process of dental erosion, although associations were moderate and weak, respectively.