Antimicrobial effect of two fluoride-releasing adhesive tapes on Streptococcus mutans biofilm.

Fluoride-releasing adhesive tapes have been developed as a new fluoride delivery agent. However, application as caries prevention agents remains underexplored. This study aimed at evaluating the antimicrobial activity of two fluoride-releasing adhesive tapes against S. mutans biofilm. Two polyvinyl alcohol (PVA) tapes were investigated: (i) a fluoride-PVA (F-PVA) tape, (ii) a pullulan incorporated F-PVA (PF-PVA) tape. S. mutan strains were cultured and treated with the tapes. Antimicrobial effects were evaluated using the agar diffusion test, field-emission scanning electron microscopy (FE-SEM), and confocal laser scanning microscopy (CLSM). F-PVA tapes showed higher inhibition-zone diameters than PF-PVA at 48 h and 72 h. However, there were no significant differences (p > 0.05) between the effects of F-PVA and PF-PVA. The bio-volume of S. mutans and extracellular polymeric substances significantly decreased in the F-PVA tapes than in the PF-PVA tapes (p < 0.05). FE-SEM micrographs revealed less S. mutans colonization in F-PVA. F-PVA exhibited better antimicrobial activity against S. mutans than PF-PVA.

Epigallocatechin-3-gallate attenuates fluoride induced apoptosis via PI3K/FoxO1 pathway in ameloblast-like cells.

Fluoride is a double-edged sword. It was widely used for early caries prevention while excessive intake caused a toxicology effect, affected enamel development, and resulted in dental fluorosis. The study aimed to evaluate the protective effect and mechanism of Epigallocatechin-3-gallate (EGCG) on the apoptosis induced by fluoride in ameloblast-like cells. We observed that NaF triggered apoptotic alterations in cell morphology, excessive NaF arrested cell cycle at the G1, and induced apoptosis by up-regulating Bax and down-regulating Bcl-2. NaF activated the insulin-like growth factor receptor (IGFR), and phosphatidylinositol-3-hydroxylase (p-PI3K), while dose-dependently down-regulating the expression of Forkhead box O1 (FoxO1). EGCG supplements reversed the changes in LS8 morphology, the cell cycle, and apoptosis induced by fluoride. These results indicated that EGCG possesses a protective effect against fluoride toxicity. Furthermore, EGCG suppressed the activation of p-PI3K and the down-regulation of FoxO1 caused by fluoride. Collectively, our findings suggested that EGCG attenuated fluoride-induced apoptosis by inhibiting the PI3K/FoxO1 signaling pathway. EGCG may serve as a new alternative method for dental fluorosis prevention, control, and treatment.

Antibacterial and oral tissue effectiveness of a mouthwash with a novel active system of amine + zinc lactate + fluoride.

OBJECTIVES: Reflecting the need for an effective support for the daily oral hygiene routine of patients experiencing (symptoms of) gum inflammation, a new mouthwash has been developed containing an amine + zinc lactate + fluoride system. The in vitro efficacy of this product was assessed using traditional laboratory methods, as well as novel experimentation. MATERIALS AND METHODS: This mouthwash has been evaluated in a series of laboratory tests including two short interval kill tests (SIKTs), a 12-h (longer term) biofilm regrowth assay, a plaque glycolysis assay, and an aerobic, repeated exposure biofilm model, as well as tests for soft tissue uptake and LPS neutralization. RESULTS: Several laboratory studies demonstrate that a mouthwash containing an amine + zinc lactate + fluoride system provides short-term and long-term antibacterial activity. While the immediate efficacy of this formula has been shown to be driven by the presence of the amine, zinc lactate provides a long-term antibacterial effect, as well as is able to inhibit bacterial metabolism. CONCLUSIONS: This research provides the basis for understanding the mode of action of this new mouthwash formulation and explains the previously observed clinical efficacy of this formula against plaque and gingivitis.

Colloidal chitosan-silver nanoparticles-fluoride nanocomposite as an antibacterial mouthwash against salivary Streptococcus mutans in orthodontic patients (a randomized clinical trial).

OBJECTIVES: This study aimed to synthesize and characterize colloidal chitosan-silver nanoparticles-fluoride nanocomposite (CCAgNPF) and evaluate its efficacy compared to chlorhexidine on salivary Streptococcus mutans in orthodontic patients. MATERIALS AND METHODS: AgNPs stabilized with chitosan were synthesized by chemical reduction of AgNO3. The nanoparticles were characterized with SEM, FTIR, DLS and ICP-OES. The MIC and MBC against S. mutans and IC50 concentration of CCAgNPF were obtained for antibacterial and cytotoxicity evaluations, respectively. For the clinical study, a total of 45 orthodontic patients were divided into three groups of 15 and used the following mouthwashes twice a day for 1 month: CCAgNPF, chlorhexidine 0.2% and the combination of these mouthwashes. The colony count of salivary S. mutans was evaluated before and after using the mouthwashes. The data were analyzed using One-way ANOVA and Tukey's test. RESULTS: Stabilized AgNPs were spherical with a diameter of 25.3 ± 3.3 nm. The MIC, MBC and IC50 of CCAgNPF were 4.42, 8.85 and 18.89 µg/ml. All mouthwashes reduced the salivary S. mutans of the orthodontic patients, however, no significant difference was found between the efficacy of CCAgNPF and chlorhexidine (P-value > 0.05). The best results were achieved by the combination of CCAgNPF and chlorhexidine mouthwashes (P-value < 0.05). CONCLUSION: The CCAgNPF and its combination with chlorhexidine present potent bactericidal, biocompatible and effective anti-carious mouthwashes for orthodontic patients. CLINICAL RELEVANCE: This study proved CCAgNPF as an antibacterial mouthwash with lower cytotoxicity and side effects for patients undergoing orthodontic treatments to maintain oral hygiene and reduce salivary S. mutans.

In vitro effect of TiF4/NaF solution on the development of radiation-induced dentin caries.

OBJECTIVE: To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions. METHODOLOGY: bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters. RESULTS: All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study.

Effect of training on bone quality in rats with insulin resistance induce by fluoride consumption.

When large amounts of Fluoride are consumed produces insulin resistance, but exercise can reverse insulin resistance in rats, because of a high fluoride uptake by bone tissue. However, bone quality has not been studied in those experiments. Therefore, the aim of this work was to evaluate bone quality in rats treated with fluoride when performing exercise. Sprague-Dawley rats were divided into 3 groups (n=6 per group): Control (drinking water without fluoride), Fluoride (drinking water with fluoride 15 mg/L for 30 days) and Exercise (daily running on a treadmill and drinking water with fluoride 15 mg/L for 30 days).  Then, bone mineral density, mechanical and histological properties and bone fluoride level were measured. No effect of treatment on any bone parameters were observed. These results indicate that exercise normalizes glucose metabolism in insulin-resistant rats by bone fluoride uptake; however, this increase in bone fluoride does not manifest in bone deterioration.

Cuando se consumen grandes cantidades de fluoruro se produce resistencia a la insulina, pero la realización de ejercicio puede revertir dicho efecto en ratas, debido a una alta absorción de fluoruro por el tejido óseo. Sin embargo, la calidad ósea no ha sido estudiada. Por ello, el objetivo de este trabajo fue evaluar la calidad ósea en ratas tratadas con flúor que realizan ejercicio. Se trabajó con ratas Sprague-Dawley que se dividieron en 3 grupos (n=6 por grupo): Control (recibiron agua sin flúor), Flúor (recibieron agua con flúor 15 mg/L durante 30 días) y Ejercicio (realizaron ejercicio diariamente en cinta ergométrica y recibieron agua con fluoruro 15 mg/L por 30 días). Luego, se midieron la densidad mineral ósea, las propiedades biomecánicas e histológicas y el nivel de fluoruro óseo. No se observó ningún efecto del tratamiento sobre ningún parámetro óseo. Estos resultados indican que el ejercicio normaliza el metabolismo de la glucosa en ratas resistentes a la insulina mediante la captación ósea de fluoruro; sin embargo, este aumento del fluoruro óseo no se manifiesta en deterioro óseo.

Fluoride impairs vascular smooth muscle A7R5 cell lines via disrupting amino acids metabolism.

Given the insidious and high-fatality nature of cardiovascular diseases (CVDs), the emergence of fluoride as a newly identified risk factor demands serious consideration alongside traditional risk factors. While vascular smooth muscle cells (VSMCs) play a pivotal role in the progression of CVDs, the toxicological impact of fluoride on VSMCs remains largely uncharted. In this study, we constructed fluorosis model in SD rats and A7R5 aortic smooth muscle cell lines to confirm fluoride impaired VSMCs. Fluoride aggravated the pathological damage of rat aorta in vivo. Then A7R5 were exposed to fluoride with concentration ranging from 0 to 1200 µmol/L over a 24-h period, revealing a dose-dependent inhibition of cell proliferation and migration. The further metabolomic analysis showed alterations in metabolite profiles induced by fluoride exposure, notably decreasing organic acids and lipid molecules level. Additionally, gene network analysis underscored the frequency of fluoride's interference with amino acids metabolism, potentially impacting the tricarboxylic acid (TCA) cycle. Our results also highlighted the ATP-binding cassette (ABC) transporters pathway as a central element in VSMC impairment. Moreover, we observed a dose-dependent increase in osteopontin (OPN) and α-smooth muscle actin (α-SMA) mRNA level and a dose-dependent decrease in ABC subfamily C member 1 (ABCC1) and bestrophin 1 (BEST1) mRNA level. These findings advance our understanding of fluoride as a CVD risk factor and its influence on VSMCs and metabolic pathways, warranting further investigation into this emerging risk factor.

Dual-species biofilm of Streptococcus mutans and Candida albicans produces subsurface caries lesions on bovine enamel.

OBJECTIVES: To develop a protocol for forming subsurface caries lesions on bovine enamel by dual-species biofilms of Streptococcus mutans and Candida albicans in vitro. DESIGN: Biofilms were grown on bovine enamel specimens in artificial saliva (AS) for seven days. After 24 h of formation, the AS was supplemented or not with fluoride (F) using sodium fluoride (0.005 or 0.008 ppm F), and the biofilms were exposed or not to a 20 % sucrose solution (reproducing a cariogenic challenge) once/day. On the seventh day, the biofilms were harvested and had their extracellular polysaccharides (EPS) and inorganic components analyzed. The specimens were subjected to computed X-ray microtomography analysis to determine their mineral concentration. Data were compared using two-way analyses of variance, followed by Fisher's LSD or Student-Newman-Keuls tests (p < 0.05). RESULTS: Biofilms exposed to the cariogenic challenge had significantly higher EPS concentrations than those not exposed, regardless of the presence of F. For biofilms grown with 0.008 ppm F, those exposed to the cariogenic challenge had lower F levels than those not exposed. For biofilms exposed to the cariogenic challenge, those grown with 0.008 ppm F had lower lesion depths and integrated mineral loss, and higher outer layers than those grown without F. CONCLUSIONS: The dual biofilm model assessed was able to create subsurface caries lesions in bovine enamel in vitro, which was influenced by the presence of F in the culture medium and exposure to sucrose.

Enhanced anti-biofilm and anti-caries potential of arginine combined with calcium glycerophosphate and fluoride.

OBJECTIVE: The aim of this work was to evaluate the antibiofilm and anticaries properties of the association of arginine (Arg) with calcium glycerophosphate (CaGP) and fluoride (F). METHODS: An active attachment, polymicrobial biofilm model obtained from saliva and bovine teeth discs were used. After the initial biofilm growth period, the enamel discs were transferred to culture medium. The treatment solutions were added to the culture media to achieve the desired final concentration. The following groups were used: negative control (Control); F (110 ppm F); CaGP (0.05 %); Arg (0.8 %) and their associations (F + CaGP; Arg + F; Arg + CaGP; Arg +F + CaGP). The following analyses were carried out: bacterial viability (total bacteria, aciduric bacteria and mutans streptococci), pH assessment of the spent culture medium, dry weight quantification, evaluation of surface hardness loss (%SH) and subsurface mineral content. Normality and homoscedasticity were tested (Shapiro-Wilk and Levene's test) and the following tests were applied: two-way ANOVA (acidogenicity), Kruskall-Wallis (microbial viability) and one way ANOVA (dry weight, %SH, mineral content). RESULTS: The association Arg + F + CaGP resulted in the lowest surface hardness loss in tooth enamel (-10.9 ± 2.3 %; p < 0.05). Arg +F + CaGP exhibited highest values of subsurface mineral content (10.1 ± 2.9 gHAP/cm3) in comparison to Control and F (p < 0.05). In comparison to Control and F, Arg +F + CaGP promoted the highest reduction in aciduric bacteria and mutans streptococci (5.7 ± 0.4; 4.4 ± 0.5 logCFU/mL, p < 0.05). CONCLUSIONS: The Arg-F-Ca association demonstrated to be the most effective combination in protecting the loss of surface hardness and subsurface mineral content, in addition to controlling important virulence factors of the cariogenic biofilm. CLINICAL SIGNIFICANCE: Our findings provide evidence that the Arg-F-Ca association showed an additive effect, particularly concerning protection against enamel demineralization. The combination of these compounds may be a strategy for patients at high risk of caries.

Antibacterial, cytotoxic and mechanical properties of a orthodontic cement with phosphate nano-sized and phosphorylated chitosan: An in vitro study.

OBJECTIVES: Evaluate, in vitro, the effect of incorporating nano-sized sodium trimetaphosphate (TMPnano) and phosphorylated chitosan (Chi-Ph) into resin-modified glass ionomer cement (RMGIC) used for orthodontic bracket cementation, on mechanical, fluoride release, antimicrobial and cytotoxic properties. METHODS: RMGIC was combined with Chi-Ph (0.25%/0.5%) and/or TMPnano (14%). The diametral compressive/tensile strength (DCS/TS), surface hardness (SH) and degree of conversion (%DC) were determined. For fluoride (F) release, samples were immersed in des/remineralizing solutions. Antimicrobial/antibiofilm activity was evaluated by the agar diffusion test and biofilm metabolism (XTT). Cytotoxicity in fibroblasts was assessed with the resazurin method. RESULTS: After 24 h, the RMGIC-14%TMPnano group showed a lower TS value (p < 0.001); after 7 days the RMGIC-14%TMPnano-0.25%Chi-Ph group showed the highest value (p < 0.001). For DCS, the RMGIC group (24 h) showed the highest value (p < 0.001); after 7 days, the highest value was observed for the RMGIC-14%TMPnano-0.25%Chi-Ph (p < 0.001). RMGIC-14%TMPnano, RMGIC-14%TMPnano-0.25%Chi-Ph, RMGIC-14%TMPnano-0.5%Chi-Ph showed higher and similar release of F (p > 0.001). In the SH, the RMGIC-0.25%Chi-Ph; RMGIC-0.5%Chi-Ph; RMGIC-14%TMPnano-0.5%Chi-Ph groups showed similar results after 7 days (p > 0.001). The RMGIC-14%TMPnano-0.25%Chi-Ph group showed a better effect on microbial/antibiofilm growth, and the highest efficacy on cell viability (p < 0.001). After 72 h, only the RMGIC-14%TMPnano-0.25%Chi-Ph group showed cell viability (p < 0.001). CONCLUSION: The RMGIC-14%TMPnano-0.25%Chi-Ph did not alter the physical-mechanical properties, was not toxic to fibroblasts and reduced the viability and metabolism of S. mutans. CLINICAL RELEVANCE: The addition of phosphorylated chitosan and organic phosphate to RMGIC could provide an antibiofilm and remineralizing effect on the tooth enamel of orthodontic patients, who are prone to a high cariogenic challenge due to fluctuations in oral pH and progression of carious lesions.

Metabolomics reveals high fructose-1,6-bisphosphate from fluoride-resistant Streptococcus mutans.

BACKGROUND: Fluoride-resistant Streptococcus mutans (S. mutans) strains have developed due to the wide use of fluoride in dental caries prevention. However, the metabolomics of fluoride-resistant S. mutans remains unclear. OBJECTIVE: This study aimed to identify metabolites that discriminate fluoride-resistant from wild-type S. mutans. MATERIALS AND METHODS: Cell supernatants from fluoride-resistant and wild-type S. mutans were collected and analyzed by liquid chromatography-mass spectrometry. Principal components analysis and partial least-squares discriminant analysis were performed for the statistical analysis by variable influence on projection (VIP > 2.0) and p value (Mann-Whitney test, p < 0.05). Metabolites were assessed qualitatively using the Human Metabolome Database version 2.0 ( http://www.hmdb.ca ), or Kyoto Encyclopedia of Genes and Genomes ( http://www.kegg.jp ), and Metaboanalyst 6.0 ( https://www.metaboanalyst.ca ). RESULTS: Fourteen metabolites differed significantly between fluoride-resistant and wild-type strains in the early log phase. Among these metabolites, 5 were identified. There were 32 differential metabolites between the two strains in the stationary phase, 13 of which were identified. The pyrimidine metabolism for S. mutans FR was matched with the metabolic pathway. CONCLUSIONS: The fructose-1,6-bisphosphate concentration increased in fluoride-resistant strains under acidic conditions, suggesting enhanced acidogenicity and acid tolerance. This metabolite may be a promising target for elucidating the cariogenic and fluoride resistant mechanisms of S. mutans.

Effect of treatment with phosphate, casein phosphopeptide and fluoride on the remineralization: in vitro study.

This study aimed to evaluate in vitro the effect protocols and anticaries agents containing casein amorphous calcium fluoride phosphopeptide-phosphate (CPP-ACPF, MI Paste Plus), sodium trimetaphosphate (TMP) and fluoride (F), in remineralization of caries lesions. Bovine enamel blocks with initial caries lesions were divided into groups (n = 12): 1) Toothpaste without F-TMP-MI Plus (Placebo); 2) Toothpaste 1100 ppm F (1100F), 3) 1100F + MI Paste Plus (1100F-MI Paste Plus), 4) Toothpaste with 1100F + Neutral gel with 4,500 ppm F + 5%TMP (1100F + Gel TMP) and 5) Toothpaste with 1100F + Neutral gel with 9,000 ppm F (1100F + Gel F). For the 4 and 5 groups the gel was applied only once for 1 minute, initially to the study. For the 3 group, after treatment with 1100F, MI Paste Plus was applied 2x/day for 3 minute. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); profile and depth of the subsuperficial lesion (PLM); concentrations of F, calcium (Ca) and phosphorus (P) in enamel was determined. The data were analyzed by ANOVA (1-criterion) and Student-Newman-Keuls test (p < 0.001). Treatment with 1100F alone led to ~ 28% higher remineralization when compared to treatment with 1100F associated with MI Paste Plus (p < 0.001). The 1100F and 1100F + Gel F groups showed similar values for %SHR (p = 0.150). 1100F + Gel TMP treatment also remineralized the enamel surface by ~ 30% and 20% when compared to the 1100F + Gel F and 1100F groups (p < 0.001). The lower lesion depth (ΔKHN) was observed for the 1100F + Gel TMP group (p < 0.001), where it was 54% and 44% lower in comparison to the 1100F and 1100F + Gel F groups (p < 0.001). Polarized light microscopy photomicrographs showed subsurface lesions in all groups, but these lesions were present to a lower extent in the 1100F + Gel TMP group (p < 0.001). Treatment with 1100F + Gel TMP promoted an increase in the concentration of Ca in the enamel by ~ 57% and ~ 26% when compared to the 1100F and 1100F + MI Paste Plus groups (p < 0.001), respectively. There were no significant differences between the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001). Similar values of P in the enamel were observed in the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001), except for the 1100F + Gel TMP group, which presented a high concentration (p < 0.001). We conclude that the 1100F+TMP gel treatment/protocol led to a significant increased remineralization when compared to the other treatments/protocols and may be a promising strategy for patients with early caries lesions.

Effect of fluoride or chitosan toothpaste and at-home bleaching in enamel roughness, tooth color, and staining susceptibility.

PURPOSE: To evaluate how fluoride- or chitosan-based toothpaste used during at-home bleaching affects enamel roughness, tooth color, and staining susceptibility. METHODS: Bovine enamel blocks were submitted to a 14-day cycling regime considering a factorial design (bleaching agent x toothpaste, 2 x 3), with n=10: (1) bleaching with 16% carbamide peroxide (CP) or 6% hydrogen peroxide (HP), and (2) daily exposure of a fluoride (1,450 ppm F-NaF) toothpaste (FT), chitosan-based toothpaste (CBT), or distilled water (control). Then, 24 hours after the last day of bleaching procedure the samples were exposed to a coffee solution. Color (ΔEab, ΔE00, L*, a*, b*) and roughness (Ra, µm) analyses were performed to compare the samples initially (baseline), after bleaching, and after coffee staining. The results were evaluated by linear models for repeated measures (L*, a*, b*, and Ra), 2-way ANOVA (ΔEab, ΔE00) and Tukey's test (α= 0.05). RESULTS: After the at-home bleaching procedure (toothpaste vs. time, P< 0.0001), the toothpaste groups presented a statistically lower Ra than the control (CBT 0.05). After coffee exposure, CBT presented lower ΔEab and ΔE00 values in the HP groups (toothpaste, P< 0.0001), and lower b* and a* values in the CP groups (toothpaste vs. time, P= 0.004). CLINICAL SIGNIFICANCE: Fluoride or chitosan delivered by toothpaste can reduce surface alterations of the enamel during at-home bleaching, without affecting bleaching efficacy.

Antibacterial effects of silver diamine fluoride, potassium iodide and nanosilver fluoride on dual-species biofilm.

OBJECTIVES: This study aims to evaluate antibacterial effects of silver diamine fluoride (SDF), SDF/potassium iodide (KI), and nanosilver fluoride (NSF). METHODS: Antimicrobial activity of sterile saline, 5% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX), SDF, SDF/KI, NSF, and KI solutions against Streptococcus mutans and Lactobacillus casei was assessed through disc diffusion tests. A dual-species biofilm of S. mutans-L. casei was formed on 48 enamel samples, divided into six groups (n = 8). Group 1 was treated with sterile saline, Group 2 with 5% NaOCl, Group 3 with 2% CHX, Group 4 with SDF, Group 5 with SDF/KI, and Group 6 with NSF. The samples were analysed using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Statistical analysis utilized Shapiro-Wilk and Kruskal-Wallis tests and multiple comparisons were conducted using Dunn test. RESULTS: SDF, SDF/KI, and NaOCl displayed significantly higher antibacterial activity against dual-species biofilm compared to NSF and CHX (p < 0.050). CONCLUSIONS: In conclusion, SDF and SDF/KI demonstrated greater antibacterial activity than NSF. SDF's antibacterial activity was unaffected by KI. Further research is needed to determine the appropriate content and concentration for achieving effective antibacterial activity with NSF. CLINICAL SIGNIFICANCE: The use of silver-containing materials is increasing in popularity within pediatric dentistry. In this study, an endeavor has been made to assist pediatric dentists in determining which solution might be more advantageous for preventing caries.

Supragingival dental biofilm profile and biofilm control during orthodontic treatment with fixed orthodontic appliance: A randomized controlled trial.

OBJECTIVE: The effectiveness of supragingival dental biofilm control during orthodontic treatment and changes in the bacterial profile were analyzed. DESIGN: Sixty-four participants aged 12-22 years (57% female) were included in the study. Participants underwent orthodontic treatment with fixed appliances and were randomly assigned to one of the three groups, which during a period of one month: (I) used chlorhexidine digluconate (CHX), (II) used high concentration of fluoride (F) gel and (III) performed standard oral hygiene. The plaque and gingivitis index, pH of biofilm and white spot lesions (WSL) were assessed. Changes of the bacteria in the biofilm were analyzed by the quantitative polymerase chain reaction RESULTS: Increase in the plaque index, pH of biofilm, and WSL was observed during orthodontic treatment with standard oral hygiene. Large interindividual variability was present, and the effects of one-month use of fluorides and CHX on clinical parameters were not significant. Despite standard hygiene the abundance of studied biofilm bacteria increased - the most Streptoccocus mutans (14.2x) and S. salivarius (3.3x), moderate Veillonella parvula (3x) and the least S. sobrinus (2.3x) and Agregatibacter actinomycetemcomitans (1.9x). The use of CHX reduced S. sobrinus (2.2x) and A. actinomycetemcomitans (1.9x). Fluoride use reduced A. actinomycetemcomitans (1.3x) and S. sobrinus (1.2x). Fluorides better controlled S. mutans than CHX. CONCLUSION: Bacterial biomass in supragingival biofilm increased during treatment with metal orthodontic appliances, with greater increase in cariogenic bacteria than periopathogens. Fluoride controlled S. mutans, while CHX S. sobrinus and A. actinomycetemcomitans.

Influence of fluoride characteristics on tooth surface protection in an erosive condition: A multifaceted characterization approach.

OBJECTIVE: To evaluate the effect of fluoride consistency and composition to protect enamel and dentin against the dental erosion. MATERIALS AND METHODS: Bovine enamel and dentin specimens were treated with artificial saliva, neutral fluoride gel (NFG), acidulated phosphate fluoride gel (AFG), neutral fluoride foam (NFF), and acidulated phosphate fluoride foam. The samples were subjected to cycling. Micro energy-dispersive X-ray fluorescence spectrometry, surface roughness (Ra), contact angle (CA), and scanning electron microscopy (SEM) were performed. Composition, CA and Ra data were analyzed by ANOVA and multiple comparison test (p < 0.05). RESULTS: The dentin protected had a significantly higher mineral content than in the control. Eroded unprotected enamel had higher Ra values than normal surfaces. Fluoride treatments increased the Ra in dentin samples. AFG increased the CA in enamel. Fluoride foams increased CA in dentin with reduced mineral loss. SEM analysis found a deposited layer on enamel treated with AFG and remnants of deposits on dentin treated with NFG and NFF. CONCLUSION: Regardless of the form of application, fluoride provided protection against erosion, however with different levels. CLINICAL SIGNIFICANCE: Applying the adequate fluoride form is relevant since the formulations have different effects on both enamel and dentin.

Low-fluoride gels supplemented with nano-sized sodium trimetaphosphate reduce dentin erosive wear in vitro.

OBJECTIVE: The study assessed the effect of low-fluoride gels supplemented with micrometric or nano-sized sodium trimetaphosphate (TMP) on dentin erosive wear in vitro. DESIGN: Bovine dentin blocks (n = 154) were selected by surface microhardness and randomly allocated into seven groups (n = 22/group), according to the gels: Placebo; 4500 ppm F (4500F); 9000 ppm F (9000F); 5% TMP microparticulate plus 4500F (5TMPm+4500F); 2.5% TMP nanoparticulate plus 4500 F (2.5TMPn+4500F); 5% TMP nanoparticulate plus 4500F (5TMPn+4500F); and 12,300 ppm F acid gel (APF). All blocks were treated only once for 60 s and cyclically eroded (ERO, citric acid, 4 × 90 s/day) or eroded and brushed (4 × 15 s/day, five strokes/s, ERO+ABR) over five days (each subgroup n = 11). Dentin wear and integrated hardness loss in depth (ΔKHN) were determined, and the data were submitted to two-way ANOVA, followed by Tukey's test, and Spearman's correlation (p < 0.05). RESULTS: For ERO, all gels containing 4500F supplemented with TMP significantly reduced dentin wear compared with their counterpart without TMP, reaching values similar to 9000F. For ERO+ABR, 5TMPn+ 4500F gel led to significantly lower wear than all its counterparts, reaching values similar to 9000F and APF. As for ΔKHN, all gels containing TMP promoted superior protective effects compared with 4500F, reaching values similar to 9000F and APF under both challenges. A positive correlation between dentin wear and mineral content in depth was verified. CONCLUSIONS: Gels containing 4500F supplemented with TMP significantly reduced dentin erosive wear compared with pure 4500F, with additional benefit from the use of nanoparticles.

Fluoride export is required for the competitive fitness of pathogenic microorganisms in dental biofilm models.

Microorganisms resist fluoride toxicity using fluoride export proteins from one of several different molecular families. Cariogenic species Streptococcus mutans and Candida albicans extrude intracellular fluoride using a CLCF F-/H+ antiporter and FEX fluoride channel, respectively, whereas oral commensal eubacteria, such as Streptococcus gordonii, export fluoride using a Fluc fluoride channel. In this work, we examine how genetic knockout of fluoride export impacts pathogen fitness in single-species and three-species dental biofilm models. For biofilms generated using S. mutans with the genetic knockout of the CLCF transporter, exposure to low fluoride concentrations decreased S. mutans counts, synergistically reduced the populations of C. albicans, increased the relative proportion of oral commensal S. gordonii, and reduced properties associated with biofilm pathogenicity, including acid production and hydroxyapatite dissolution. Biofilms prepared with C. albicans with genetic knockout of the FEX channel also exhibited reduced fitness in the presence of fluoride but to a lesser degree. Imaging studies indicate that S. mutans is highly sensitive to fluoride, with the knockout strain undergoing complete lysis when exposed to low fluoride for a moderate amount of time. Biochemical purification of the S. mutans CLCF transporter and functional reconstitution establishes that the functional protein is a dimer encoded by a single gene. Together, these findings suggest that fluoride export by oral pathogens can be targeted by specific inhibitors to restore biofilm symbiosis in dental biofilms and that S. mutans is especially susceptible to fluoride toxicity. IMPORTANCE: Dental caries is a globally prevalent condition that occurs when pathogenic species, including Streptococcus mutans and Candida albicans, outcompete beneficial species, such as Streptococcus gordonii, in the dental biofilm. Fluoride is routinely used in oral hygiene to prevent dental caries. Fluoride also has antimicrobial properties, although most microbes possess fluoride exporters to resist its toxicity. This work shows that sensitization of cariogenic species S. mutans and C. albicans to fluoride by genetic knockout of fluoride exporters alters the microbial composition and pathogenic properties of dental biofilms. These results suggest that the development of drugs that inhibit fluoride exporters could potentiate the anticaries effect of fluoride in over-the-counter products like toothpaste and mouth rinses. This is a novel strategy to treat dental caries.

Novel pit and fissure sealant with nano-CaF2 and antibacterial monomer: Fluoride recharge, microleakage, sealing ability and cytotoxicity.

Conventional resin-based sealants release minimal fluoride ions (F) and lack antibacterial activity. The objectives of this study were to: (1) develop a novel bioactive sealant containing calcium fluoride nanoparticles (nCaF2) and antibacterial dimethylaminohexadecyl methacrylate (DMAHDM), and (2) investigate mechanical performance, F recharge and re-release, microleakage, sealing ability and cytotoxicity. Helioseal F served as commercial control. The initial F release from sealant containing 20% nCaF2 was 25-fold that of Helioseal F. After ion exhaustion and recharge, the F re-release from bioactive sealant did not decrease with increasing number of recharge and re-release cycles. Elastic modulus of new bioactive sealant was 44% higher than Helioseal F. The new sealant had excellent sealing, minimal microleakage, and good cytocompatibility. Hence, the nanostructured sealant had substantial and sustained F release and antibacterial activity, good sealing ability and biocompatibility. The novel bioactive nCaF2 sealant is promising to provide long-term F ions for caries prevention.

Measurement and analysis of microbial fluoride resistance in dental biofilm models.

The interactions between communities of microorganisms inhabiting the dental biofilm is a major determinant of oral health. These biofilms are periodically exposed to high concentrations of fluoride, which is present in almost all oral healthcare products. The microbes resist fluoride through the action of membrane export proteins. This chapter describes the culture, growth and harvest conditions of model three-species dental biofilm comprised of cariogenic pathogens Streptococcus mutans and Candida albicans and the commensal bacterium Streptococcus gordonii. In order to examine the role of fluoride export by S. mutans in model biofilms, procedures for generating a strain of S. mutans with a genetic knockout of the fluoride exporter are described. We present a case study examining the effects of this mutant strain on the biofilm mass, acid production and mineral dissolution under exposure to low levels of fluoride. These general approaches can be applied to study the effects of any gene of interest in physiologically realistic multispecies oral biofilms.