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.

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.

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.

Spatial mapping of polymicrobial communities reveals a precise biogeography associated with human dental caries.

Tooth decay (dental caries) is a widespread human disease caused by microbial biofilms. Streptococcus mutans, a biofilm-former, has been consistently associated with severe childhood caries; however, how this bacterium is spatially organized with other microorganisms in the oral cavity to promote disease remains unknown. Using intact biofilms formed on teeth of toddlers affected by caries, we discovered a unique 3D rotund-shaped architecture composed of multiple species precisely arranged in a corona-like structure with an inner core of S. mutans encompassed by outer layers of other bacteria. This architecture creates localized regions of acidic pH and acute enamel demineralization (caries) in a mixed-species biofilm model on human teeth, suggesting this highly ordered community as the causative agent. Notably, the construction of this architecture was found to be an active process initiated by production of an extracellular scaffold by S. mutans that assembles the corona cell arrangement, encapsulating the pathogen core. In addition, this spatial patterning creates a protective barrier against antimicrobials while increasing bacterial acid fitness associated with the disease-causing state. Our data reveal a precise biogeography in a polymicrobial community associated with human caries that can modulate the pathogen positioning and virulence potential in situ, indicating that micron-scale spatial structure of the microbiome may mediate the function and outcome of host-pathogen interactions.

Teaching Tooth Wear Diagnosis, Risk Assessment, and Treatment Decisions Applying Inverted Classroom Combined with Case Presentation Model.

This study explored the application of a model for teaching tooth wear diagnosis, risk assessment, and treatment decisions to undergraduates at the UCLouvain, Belgium, based on an inverted classroom combined with case presentations. The aim was to explain its implementation and assess improvement in learning, engagement, and satisfaction. The hypothesis tested was that this model would enhance students' performance. This controlled clinical trial included 29 dental students in the test group and 30 in the control group. All students received instructions and pre-class material for reading via e-mail 2 weeks prior to class time (T0). The test group included students attending the class time (attendance was not obligatory). The control group consisted of students only attending the preclinical training (attendance was obligatory). Both groups assessed three case presentations with no, moderate, and severe erosive tooth wear. The test group assessed the cases at the beginning of the class time (T1). One week later, at the beginning of the preclinical training, the students of the control group assessed the case presentations (T2). In parallel, to measure the contribution of the class time to students' performance, the test group reassessed the cases (T2). This was followed by discussion of each case in which the lecturer presented the benchmark assessments. Students' perception of their learning experience was recorded. The generalized linear mixed regression model showed that for the overall assessments of cases, students in the control group were significantly less likely to agree with the benchmark than students in the test group (OR = 0.62; p = 0.006). Students' satisfaction was higher in the test group than in the control group but only significantly regarding the quality of the cases presentations (Fisher test p < 0.01). The class time contributed significantly to students' improvement in learning in the test group (liner mixed model; p < 0.01). In conclusion, the model applied improved significantly in performance, learning, and to some extent satisfaction. However, the motivation strategy applied resulted in only half of the students adhering/engaging to the complete interactive model of teaching. Further motivation strategies should be implemented to make the complete model more widely accepted by students.

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.

Ferumoxytol Nanoparticles Target Biofilms Causing Tooth Decay in the Human Mouth.

Severe tooth decay has been associated with iron deficiency anemia that disproportionally burdens susceptible populations. Current modalities are insufficient in severe cases where pathogenic dental biofilms rapidly accumulate, requiring new antibiofilm approaches. Here, we show that ferumoxytol, a Food and Drug Administration-approved nanoparticle formulation for treating iron deficiency, exerts an alternative therapeutic activity via the catalytic activation of hydrogen peroxide, which targets bacterial pathogens in biofilms and suppresses tooth enamel decay in an intraoral human disease model. Data reveal the potent antimicrobial specificity of ferumoxytol iron oxide nanoparticles (FerIONP) against biofilms harboring Streptococcus mutans via preferential binding that promotes bacterial killing through in situ free-radical generation. Further analysis indicates that the targeting mechanism involves interactions of FerIONP with pathogen-specific glucan-binding proteins, which have a minimal effect on commensal streptococci. In addition, we demonstrate that FerIONP can detect pathogenic biofilms on natural teeth via a facile colorimetric reaction. Our findings provide clinical evidence and the theranostic potential of catalytic nanoparticles as a targeted anti-infective nanomedicine.

Diagnosis, risk assessment, and treatment decisions for tooth wear in daily practice: a case presentation survey among Belgian dentists.

This survey assessed the use of current concepts for tooth wear diagnosis, risk assessment, and treatment decisions in daily practice, and identified factors of significance for treatment decisions. A total of 104 dentists were presented with 10 cases representing varying degrees of tooth wear. The dentists were asked to answer six questions pertaining to the diagnosis, risk assessment, and treatment of each case. A benchmark was established by three experts and the agreement between dentists and the benchmark was calculated. At dentition level, the agreement was moderate for diagnosis (κ = 0.55) and risk assessment (κ = 0.54). At tooth/surface level, the agreement for clinical and radiographic severity of wear was moderate (κ = 0.57) and substantial (κ = 0.65), respectively. The agreement for treatment decision was fair (κ = 0.35). Multivariate logistic regression analysis showed that treatment decisions for operative treatment were associated with moderate to high case risk (OR = 10.1; 95% CI: 5.4-18.9). Belgian dentists performed reasonably well in applying current concepts and strategies for tooth wear diagnosis and risk assessments at dentition level, as well as for assessment of wear severity at tooth/surface level. Improvement is warranted concerning treatment decisions. This survey could be a useful tool for disseminating and improving current knowledge of tooth wear, and for training dentists in daily practice.

Tooth Age Impact on Dental Erosion Susceptibility and Treatment Efficacy.

This laboratory study investigated the impact of tooth age on dental erosion susceptibility and preventive treatment efficacy. Extracted human premolars were selected and had their age estimated (∼10-100 years old) using established dental forensic methods. Enamel and root dentin slabs were prepared, embedded in acrylic blocks, flattened, and polished. The specimens were randomly assigned to one of three treatments (n = 93): Sn+F (800 ppm Sn as SnCl2 and 250 ppm F as NaF, pH 4.5), NaF (250 ppm F, pH 4.5), or deionized water (DIW). Each specimen was subjected for 10 days to a daily cycling protocol consisting of six 5-min erosive challenges (0.3% citric acid, pH 2.6), six 60-min remineralization periods (artificial saliva), and three 2-min treatments with the test solutions. Surface loss (SL) was measured after 3, 5, and 10 days, using optical profilometry. Effects of tooth age, antierosive treatment, and time on SL were evaluated using linear mixed effects regression analysis. SL increased with age for all substrate-treatment-time combinations (p < 0.0001). Sn+F and NaF solutions significantly reduced SL compared to DIW, regardless of substrate, time, or age (p < 0.0001), with best results shown for Sn+F. Efficacy of Sn+F increased with tooth age on enamel, but tooth age did not affect the efficacy of NaF on enamel. For dentin, increased efficacy was observed with age after 5 (for Sn+F) and 10 days (for Sn+F and NaF). In conclusion, increase of tooth age rendered enamel and root dentin more susceptible to dental erosion. NaF preventive efficacy improved with tooth age for dentin, in advanced erosion simulation. Sn+F reduced enamel SL due to erosion regardless of tooth age.

Protective Effect of Solutions Containing Polymers Associated with Fluoride and Stannous Chloride on Hydroxyapatite Dissolution.

This study investigated the protective effect of experimental solutions containing 4 polymers (polyoxirane, hydroxypropylmethylcellulose [HPMC], pectin, and an amino methacrylate copolymer [AMC]) in 2 concentrations (low and high) associated or not with sodium fluoride (F; 225 ppm F-) or sodium fluoride plus stannous chloride (FS; 800 ppm Sn2+) on the dissolution of hydroxyapatite crystals (HA). Deionized water was the control. The pretreated HA was added to a 0.3% citric acid solution (pH 3.8). An automatic titrant machine added aliquots of 0.1 N HCl at a rate of 28 µL/min, in a total reaction time of 5 min. Groups were compared with 2-way ANOVA and Tukey's test, and concentrations with Student t test (5%). The zeta potential of the HA treated with the solutions was measured. Significant differences were found for both factors and interaction (p < 0.0001). The treatments with F and FS solutions resulted in a lower amount of dissolved HA than the control. Among the polymers' solutions, only AMC was able to reduce the amount of dissolved HA, changing the surface charge of HA to positive. AMC improved the protective effect of F, but it did not affect FS. Polyoxirane and HPMC reduced the protective potential of the FS solution. No differences were found between the concentrations of the polymers. It was concluded that F and FS reduced the amount of dissolved HA. The protective effect of the experimental solutions against HA dissolution was polymer dependent. The F effect was enhanced by its combination with AMC, but the protection of FS was impaired by polyoxirane and HPMC.

Erosive tooth wear inhibition by hybrid coatings with encapsulated fluoride and stannous ions.

This study aimed to formulate a hybrid coating material (HC) and to modify this HC with fluoride (NaF) and stannous (SnCl2) ions, directly or encapsulated in nano containers, testing the effects of these materials against dental erosion and erosion-abrasion. Enamel and dentin specimens were treated with the HCs, and then tested in erosion or erosion-abrasion cycling models of 5 days (n = 10 for each substrate, for each model). Deionized water was the negative control, and a fluoride varnish, the positive control. Surface loss (SL, in µm) was evaluated with an optical profilometer, and data were statistically analyzed (α = 0.05). For enamel, in erosion, the positive control and HC without additives showed significantly lower SL than the negative control (p = 0.003 and p = 0.001). In erosion-abrasion, none of the groups differed from the negative control (p > 0.05). For dentin, in erosion, the positive control, HC without additives, HC with non-encapsulated F, and HC with encapsulated F + Sn showed lower SL than the negative control (p < 0.05). In erosion-abrasion, none of the groups differed significantly from the negative control (p < 0.05). HC without additives showed a promising potential for protecting the teeth against dental erosion (with upward trend for improved protection on dentin), but not against erosion-abrasion. The presence of additives did not improve the protective effect of the HC, on both substrates.

Objective assessment of simulated non-carious cervical lesion by tridimensional digital scanning.

OBJECTIVE: To explore the use of 3D intraoral scanner/image analysis for the detection and monitoring of simulated non-carious cervical lesions (NCCLs) in vitro. MATERIALS AND METHODS: A total of 288 NCCLs of different severities and simulated using a laboratorial model associating toothbrush stiffness (soft, medium, and hard) and toothpaste abrasivity (low, medium, high, and negative control) were analyzed. Dental impressions were taken from specimens before and after 35K and 65K brushing strokes, and then scanned with a CEREC Omnicam scanner. 3D models were analyzed for volumetric tooth loss. 3D optical profilometry was considered as the gold standard. Data were analyzed using ANOVA and Fisher's PLSD tests (alpha = 0.05), and agreement between methods by using intraclass correlation coefficient. RESULTS: Toothbrushes of hard and mid stiffness caused higher tooth loss than soft when associated with the highest abrasive, at 35K and 65K strokes (p < 0.001). Variation in slurry abrasivity led to differences in tooth loss (with control < low < medium < high, p < 0.0001) after both 35K and 65K strokes, regardless of the type of toothbrush used, except at 35K, wherein control = low (p = 0.55). 35K strokes caused less tooth loss than 65K for all abrasive slurries (p < 0.0001) except controls. The intraclass correlation coefficient for agreement between the test and gold standard methods was 0.85. CONCLUSIONS: Analysis of 3D images from intraoral scanner could detect and monitor NCCL progression, although this ability was limited on incipient lesions. Overall good agreement was found between the test method and optical profilometry. CLINICAL RELEVANCE: The suggested method may be applicable to detect and monitor NCCLs clinically.

Susceptibility of Dental Enamel of Different Ages to Caries-Like Lesion Development.

This study investigated the impact of estimated age, anatomical location, and the presence of wear facets on the susceptibility of enamel to develop caries-like lesions. Extracted human premolars (n = 261) had their age estimated between 10 and 93 years old, using established forensic methods. Specimens of enamel (4 × 4 mm) were prepared from the middle of the buccal surfaces, preserving the outer surface layer. The central area of the block (4 × 1 mm) was protected with nail polish and used as an internal control. The specimens were demineralized for 8 days (with 0.1 M acetic acid, 1.28 mM Ca, 0.74 mM Pi, and 0.03 µg F/mL, pH 5.0), to simulate caries-like lesion development. They were then scanned individually using microtomography, and digital 2D images were used to calculate the outcomes of integrated mineral concentration loss (ΔZ in µm/g/cm3) and lesion depth (LD in µm) at 3 locations, i.e., the cervical, middle, and occlusal thirds. The presence of natural surface wear facets was considered in the analysis. Data were evaluated using a linear mixed-effects models (α = 0.05). ΔZ increased significantly as a function of estimated tooth age at all 3 locations, and this increase was greater after the age of 30 years (p < 0.001), when a higher ΔZ was found in the occlusal third than in the middle and cervical thirds (p < 0.001). LD increased only in the occlusal third before the age of 30 years (p = 0.039) and this increase was significantly greater after 30 years at all 3 locations (p < 0.01), with no differences among them (p > 0.15). The presence of wear facets significantly increased ΔZ and LD (p < 0.001 for both). Overall, we concluded that the susceptibility of enamel to developing caries-like lesions increased with estimated dental age. This effect was more pronounced after the estimated age of 30 years and in the presence of natural tooth wear facets.

Terminology of Erosive Tooth Wear: Consensus Report of a Workshop Organized by the ORCA and the Cariology Research Group of the IADR.

Our understanding of erosive tooth wear and its contributing factors has evolved considerably over the last decades. New terms have been continuously introduced, which frequently describe the same aspects of this condition, whereas other terms are being used inappropriately. This has led to unnecessary confusion and miscommunication between patients, professionals, and researchers. A group of 15 experts, selected by the European Organization for Caries Research (ORCA) and the Cariology Research Group of the International Association for Dental Research (IADR), participated in a 2-day workshop to define the most commonly used terms in erosive tooth wear. A modified Delphi method was utilized to reach consensus. At least 80% agreement was achieved for all terms discussed and their definitions related to clinical conditions and processes, basic concepts, diagnosis, risk, and prevention and management of erosive tooth wear. Use of the terms agreed on will provide a better understanding of erosive tooth wear and intends to enable improved communication on this topic.

Toothpaste factors related to dentine tubule occlusion and dentine protection against erosion and abrasion.

OBJECTIVES: To investigate the effect of toothpastes on dentine surface loss and tubule occlusion, and the association of toothpaste-related factors to each of the outcomes. MATERIALS AND METHODS: One hundred and sixty human dentine specimens were randomly distributed into 10 groups, according to different toothpastes. The specimens were submitted to artificial saliva (60 min), citric acid (3 min), and brushing abrasion (25 s; totalizing 2 min in toothpaste slurries). This was repeated five times and two outcome variables were analyzed: dentine surface loss (dSL; µm) and tubule occlusion by measurement of the total area of open tubules (Area-OT; µm2). Data were analyzed with Kruskal-Wallis and Mann-Whitney tests (α = 0.05); bivariate and multivariate regressions were used to model the association of the chemical (pH, concentration of F-, Ca2+, and PO43- and presence of Sn2+) and physical (% weight of solid particles, particle size, and wettability) factors of the toothpastes to both outcome variables. RESULTS: Toothpastes caused different degrees of dSL and did not differ in Area-OT. All chemical and physical factors, except the presence of Sn2+, were associated with dSL (p < 0.001). Area-OT was associated only with the presence of Sn2+ (p = 0.033). CONCLUSION: Greater dSL was associated with lower pH, lower concentration of F-, higher concentration of Ca2+ and PO43-, greater % weight of solid particles, smaller particle size, and lesser wettability, whereas tubule occlusion was associated with the presence of Sn2+. CLINICAL RELEVANCE: Depending on their chemical and physical composition, toothpastes will cause different degrees of dentine tubule occlusion and dentine surface loss. This could, in turn, modulate dentine hypersensitivity.

The effects of charcoal dentifrices on Streptococcus mutans biofilm development and enamel demineralization.

PURPOSE: To evaluate the in vitro effects of commercially available charcoal dentifrices on Streptococcus mutans biofilm development and their ability to prevent enamel demineralization. METHODS: Streptococcus mutans biofilm was formed on polished bovine enamel specimens (n= 9 per treatment), and treated twice-daily for 120 seconds over the course of 5 days with: charcoal dentifrice containing fluoride (1,000 ppm F) (CF+), fluoride-free charcoal dentifrice (CF-), regular fluoride (1,100 ppm F) dentifrice (F+ ), or regular fluoride-free dentifrice (F-). Chlorhexidine (CHX, 0.12%) and deionized water (DIW) were used as positive and negative controls, respectively. Biofilms were analyzed for bacterial viability (colony-forming units, CFU). The pH of the medium was measured daily. Enamel specimens were analyzed using Vickers microhardness ( HV) and transversal microradiography (TMR). Data were analyzed using one-way ANOVA followed by post-hoc tests (α= 0.05). RESULTS: F+ showed higher pH values than CF+ and CF-, and CF- presented higher pH than CF+, showing that CF+ did not have inhibitory effects on the acidogenicity of cariogenic biofilms. CFU was significantly decreased when specimens were treated with CF+, CF- and F+, compared to specimens treated with DIW (P≤ 0.035) or F- (P≤ 0.001), respectively. However, the reduction observed was minimal (approximately 1 log). CF+ and CF- were less effective than F+ in preventing enamel demineralization as determined using HV (P= 0.041 and P= 0.003, respectively) and TMR ( P≤ 0.001). Both charcoal dentifrices (CF+, CF-) did not show relevant inhibition of S. mutans biofilm growth. Additionally, neither product prevented enamel demineralization compared to a regular fluoride-containing dentifrice. CLINICAL SIGNIFICANCE: The tested charcoal dentifrices did not exhibit anticaries potential.

In situ efficacy of an experimental toothpaste on enamel rehardening and prevention of demineralisation: a randomised, controlled trial.

BACKGROUND: A novel sodium fluoride toothpaste containing lactate ion and polyvinylmethylether-maleic anhydride has been developed to promote enamel remineralisation and resistance to demineralisation. In this in situ study, we compared this toothpaste ('Test') with a stannous fluoride-zinc citrate (SnF2-Zn) toothpaste ('Reference') (both 1100-1150 ppm fluoride) and a fluoride-free toothpaste ('Placebo') using an enamel dental erosion-rehardening model. METHODS: In each phase of this randomised, investigator-blind, crossover study, participants wore palatal appliances holding bovine enamel specimens with erosive lesions. They brushed their natural teeth with either the Test, Reference or Placebo toothpastes, then swished the resultant slurry. Specimens were removed at 2 h and 4 h post-brushing and exposed to an in vitro acid challenge. Surface microhardness was measured at each stage; enamel fluoride uptake was measured after in situ rehardening. Surface microhardness recovery, relative erosion resistance, enamel fluoride uptake and acid resistance ratio were calculated at both timepoints. RESULTS: Sixty two randomised participants completed the study. Test toothpaste treatment yielded significantly greater surface microhardness recovery, relative erosion resistance and enamel fluoride uptake values than either Reference or Placebo toothpastes after 2 and 4 h. The acid resistance ratio value for Test toothpaste was significantly greater than either of the other treatments after 2 h; after 4 h, it was significantly greater versus Placebo only. No treatment-related adverse events were reported. CONCLUSIONS: In this in situ model, the novel-formulation sodium fluoride toothpaste enhanced enamel rehardening and overall protection against demineralisation compared with a fluoride-free toothpaste and a marketed SnF2-Zn toothpaste. TRIAL REGISTRATION: ClinicalTrials.gov; NCT03296072; registered September 28, 2017.

Anti-Erosive Effect of Solutions Containing Sodium Fluoride, Stannous Chloride, and Selected Film-Forming Polymers.

The aim of this study was to evaluate the anti-erosive effect of solutions containing sodium fluoride (F: 225 ppm F-), stannous chloride (Sn: 800 ppm Sn2+), and some film-forming polymers (Gantrez: Poly [methylvinylether-alt-maleic anhydride]; PGA: propylene glycol alginate; Plasdone: poly[vinylpyrrolidone]; and CMC: carboxymethylcellulose). Solutions were tested in an erosion-remineralization cycling model, using enamel and dentin specimens (n = 10, for each substrate). Distilled water was the negative control. Cycling consisted of 120 min immersion in human saliva, 5 min in 0.3% citric acid solution, and 120 min of exposure to human saliva, 4×/day, for 5 days. Treatment with solutions (pH = 4.5) was carried out 2×/day, for 2 min. Surface loss (SL) was evaluated with optical profilometry. Zeta potential of hydroxyapatite crystals was determined after treatment with the solutions. Data were statistically analyzed (α = 0.05). For enamel, all polymers showed significantly lower SL (in µm) than the control (11.09 ± 0.94), except PGA (10.15 ± 1.25). PGA significantly improved the protective effect of F (4.24 ± 0.97 vs. 5.64 ± 1.60, respectively). None of the polymers increased the protection of F+Sn (5.13 ± 0.78). For dentin, only Gantrez (11.40 ± 0.97) significantly reduced SL when compared with the negative control (12.76 ± 0.75). No polymer was able to enhance the effect of F (6.28 ± 1.90) or F+Sn (7.21 ± 1.13). All fluoridated solutions demonstrated significantly lower SL values than the control for both substrates. Treatment of hydroxyapatite nanoparticles with all solutions resulted in more negative zeta potentials than those of the control, except Plasdone, PGA, and F+Sn+PGA, the latter two presenting the opposite effect. In conclusion, Gantrez, Plasdone, and CMC exhibited an anti-erosive effect on enamel. PGA increased the protection of F. For dentin, only Gantrez reduced erosion.

Bleaching of simulated stained-remineralized caries lesions in vitro.

OBJECTIVE: Non-invasive esthetic treatment options for stained arrested caries lesions have not been explored. This study aimed to develop laboratory models to create stained-remineralized caries-like lesions (s-RCLs) and to test the efficacy of bleaching on their esthetic treatment. MATERIALS AND METHODS: One hundred twelve enamel/dentin specimens were prepared from human molars, embedded, and had their color measured spectrophotometrically at baseline and after demineralization. They were randomly divided into four groups (n = 14) based on the staining/remineralization protocols for a total of 5 days: G1, no staining/no remineralization; G2, no staining/remineralization in artificial saliva (AS); G3, non-metallic staining/remineralization with sodium fluoride/AS; and G4, metallic staining/remineralization with silver diamine fluoride/AS. The lesion mineral loss (ΔZ) and depth (L) were measured using transverse microradiography along with color change (ΔE). Specimens were bleached and color was re-evaluated. Data were analyzed using ANOVA models followed by Fisher's PLSD tests (α = 0.05). RESULTS: s-RCLs in G4 were significantly (p < 0.001) darker than G3, G2, and G1 regardless of substrate type and condition. s-RCLs in G2, G3, and G4 showed significantly lower ΔZ and L than G1 (all p < 0.001), confirming occurrence of remineralization. G4 exhibited significantly lower ΔZ and L compared to G2 (p < 0.001). Bleaching was more effective in non-metallic than in metallic stained lesions regardless of substrate type (p < 0.001). CONCLUSION: The proposed models created distinct s-RCLs. Non-metallic s-RCLs were lighter and more responsive to bleaching compared to metallic s-RCLs. CLINICAL RELEVANCE: The developed experimental models allow the further investigation of the efficacy and safety of different clinical strategies for the esthetic management of s-RCLs.

Interplay between toothbrush stiffness and dentifrice abrasivity on the development of non-carious cervical lesions.

OBJECTIVE: This study investigated the effect of toothbrush stiffness and dentifrice slurry abrasivity on the development and progression of simulated non-carious cervical lesions (NCCLs). MATERIALS AND METHODS: Human maxillary premolars were allocated to 12 groups generated by the association between toothbrushes, soft, medium, and hard stiffness, and simulated dentifrice slurries, lower, medium, and higher; deionized water (DI) served as negative control. Teeth were mounted on acrylic blocks, and their root surfaces partially covered with acrylic resin to simulate gingiva, leaving a 2-mm area apical to the cemento-enamel junction exposed to toothbrushing. Specimens were brushed with the test slurries for 35,000 and 65,000 double strokes. Impressions taken at baseline and after both brushing periods were scanned by a 3D optical profilometer. Dentin volume loss (mm3) was calculated by image subtraction. Data were analyzed using three-way ANOVA and Fisher's PLSD tests. RESULTS: All toothbrushes caused higher volume loss when associated to higher abrasive slurry, compared to medium- and lower-abrasive slurries. Medium caused more volume loss than lower-abrasive slurry, which led to more volume loss than DI. Hard and medium toothbrushes were not different when used with medium- or higher-abrasive slurries. There were no differences among toothbrushes when used with DI and lower-abrasive slurry. Overall, 35,000 brushing strokes resulted in significantly less volume loss than 65,000. CONCLUSIONS: Toothbrush stiffness was an important factor on NCCL development, especially when brushing with medium- and higher-abrasive slurries. CLINICAL RELEVANCE: Medium and hard toothbrushes associated with medium- and high-abrasive toothpastes can yield more severe NCCLs.