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.

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.

The use of mouthwash containing trimetaphosphate as an adjunct therapy to fluoridated toothpaste reduces enamel demineralization.

INTRODUCTION: The decline in dental caries has been attributed to the widespread use of fluoride (F). Two forms of presentation are fluoridated toothpaste (FT) and mouthwash (MW), widely used by the population. MATERIALS AND METHODS: This study aimed to evaluate in vitro the effects of combining FT and MW, whether supplemented with sodium trimetaphosphate (TMP) or not, on dental enamel demineralization. Bovine enamel blocks (n = 60) were selected based on initial surface hardness (SHi) and divided into 5 experimental groups (n = 12 each): I) Placebo Toothpaste (without F/TMP); II) 1100 ppm F Toothpaste (FT); III) 1100F associated with a MW at 100 ppm F (FT + MW 100F); IV) 1100F associated with a MW at 225 ppm F (FT + MW 250F); and V) 1100F associated with a MW at 100 ppm F supplemented with 0.4 % TMP (FT + MW 100F-TMP). The blocks were treated twice a day, undergoing 5 pH cycles over 7 days. Thus, the percentage change in surface hardness (%SH), integrated subsurface hardness loss (ΔKHN), and the concentration of F, phosphorus (P), and calcium (Ca) in the enamel were determined. The data were submitted to ANOVA and Student-Newman-Keuls test (p < 0.001). RESULTS: The 1100F group was statistically inferior to the groups associated with MW for %SH, ΔKHN, and the concentration of P and Ca in the enamel (p < 0.001). Blocks treated with FT + MW 225F and FT + MW 100F-TMP showed significantly lower %SH compared to the other groups (p < 0.001). The FT + MW 100F - TMP group exhibited the lowest depth mineral loss (ΔKHN), and higher concentration de P in enamel (p < 0.001). CONCLUSION: The adjunct use of MW with FT produces a greater protective effect in inhibiting enamel demineralization, and the supplementation of TMP to the MW with 100F provides a superior effect compared to MW with 225F. CLINICAL SIGNIFICANCE: This combination of treatments could be regarded as one of several alternative fluoride supplements for subjects at elevated risk of caries.

Biomineralization and remineralizing potential of toothpastes containing nanosized ß-calcium glycerophosphate: an in vitro study.

To evaluate the effect of 1100 ppm F toothpastes supplemented with micrometric or nanosized ß-CaGP (ß-CaGPm/ß-CaGPn) on artificial enamel remineralization, using a pH cycling model. Enamel blocks with artificial caries were randomly allocated into ten groups (n = 10), according to the toothpastes: without fluoride/ß-CaGPm/ß-CaGPn (negative control); 1100 ppm F (1100F); 1100F plus 0.125%, 0.25%, 0.5%, and 1.0% of ß-CaGPm or ß-CaGPn. The blocks were treated 2×/day with slurries of toothpastes. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); integrated mineral loss (ΔIMR); fluoride (F), calcium (Ca), and phosphorus (P) concentrations in the enamel; polydispersity index (PdI); and zeta potential (Zp) were determined. The data were analyzed by ANOVA (p < 0.001). For Zp/PdI, no significance was observed when comparing the means (p > 0.001). The treatment with 1100F-0.25%ß-CaGPn led to %SHR ∼57 higher when compared to the 1100F group (p < 0.001). The lowest ΔKHN was observed for the 1100F-0.25%ß-CaGPn group (p < 0.001). The ΔIMR was lower (∼201%) for the 1100F-0.25%ß-CaGPn when compared to 1100F (p < 0.001). The association of ß-CaGPm and ß-CaGPn to 1100F did not influence its F concentration (p > 0.001). The highest increase in Ca and P was observed for 1100F-0.25%ß-CaGPn (p < 0.001). The addition of 0.25%ß-CaGPn to 1100F toothpaste was able to promote an additional remineralizing effect of artificial caries lesions.

Effect of nanometric ß-calcium glycerophosphate supplementation in conventional toothpaste on enamel demineralization: An in vitro study.

The aim of this study was to evaluate the effects of supplementing toothpastes containing 1100 ppm F with micrometric or nanometric [beta]-calcium glycerophosphate (ß-CaGPm/ß-CaGPn) on artificial enamel demineralization, using a pH cycling model. Bovine enamel blocks (4 mm × 4 mm, n = 120) selected using initial surface hardness were randomly allocated to ten toothpaste groups (n = 12): without fluoride or ß-CaGPm or ß-CaGPn (Negative control), 1100 ppm F (1100 F), and 1100 ppm F plus 0.125%, 0.25%, 0.5%, and 1.0% of ß-CaGPm or ß-CaGPn. Blocks were treated two times per day with toothpaste slurry and subjected to five pH cycles (demineralizing and remineralizing solutions) at 37 °C. The final surface hardness, percentage of surface hardness loss (%SH), cross-sectional hardness (ΔKHN), and profile analysis and lesion depth subsurface were analysed using polarized light microscopy (PLM). Fluoride (F), calcium (Ca), and phosphorus (P) concentrations were also measured. Data were analysed using ANOVA and Student-Newman-Keuls tests ([alpha] = 0.001). Blocks treated with 1100 F toothpaste containing 0.5%ß-CaGPm or 0.25%ß-CaGPn showed with reduced %SH values when compared with those treated with 1100 F alone (p < 0.001). Reduced lesion depths (ΔKHN and PLM) were observed for the slurry made up of 1100 F and 0.25%ß-CaGPn (p < 0.001). The addition of ß-CaGPm and ß-CaGPn did not influence the enamel F concentration, with the 1100 F/0.25%ß-CaGPn group exhibiting the highest Ca and P enamel concentrations (p < 0.001). Based on the findings of this in vitro study, we can conclude that the fluoride toothpaste produced a superior effect when combined at an appropriate ß-CaGP molar ratio. This effect was achieved with a lower proportion of ß-CaGP in the form of nanometric particles.

Pomegranate extract in polyphosphate-fluoride mouthwash reduces enamel demineralization.

OBJECTIVES: To evaluate the anti-demineralizing effect of a mouthwash comprising pomegranate peel extract (PPE 3%), sodium trimetaphosphate (TMP 0.3%), and fluoride (F 225 ppm) in an in situ study, and to assess its irritation potential in an ex vivo study. METHODS: This double-blind crossover study was conducted in four phases with 7 days each. Twelve volunteers used palatal appliances containing enamel blocks, which were subjected to cariogenic challenges. The ETF formulation (PPE + TMP + F, pH 7.0), TF formulation (TMP + F, pH 7.0), deionized water (W, pH 7.0), and essential oil commercial mouthwash (CM, 220 ppm F, pH 4.3) were dropped onto the enamel twice daily. The percentage of surface hardness loss, integrated loss of subsurface hardness, calcium, phosphorus, and fluoride in enamel and biofilms were determined. In addition, alkali-soluble extracellular polysaccharide concentrations were analyzed in the biofilms. The irritation potential was evaluated using the hen's egg chorioallantoic membrane test through the vascular effect produced during 300-s of exposure. RESULTS: ETF was the most efficacious in preventing demineralization. It also showed the highest concentrations of calcium and phosphorus in the enamel and in the biofilm, as well as the lowest amount of extracellular polysaccharides in the biofilm. In the eggs, ETF produced light reddening, whereas CM led to hyperemia and hemorrhage. CONCLUSIONS: The addition of PPE to formulations containing TMP and F increased its anti-demineralizing property, and this formulation presented a lower irritation potential than the CM. CLINICAL RELEVANCE: ETF can be a promising alternative alcohol-free mouthwash in patients at high risk of caries.

Influence of bleaching gels formulated with nano-sized sodium trimetaphosphate and fluoride on the physicochemical, mechanical, and morphological properties of dental enamel.

OBJECTIVES: To evaluate in vitro the effects of sodium fluoride (F) and nano-sized sodium trimetaphosphate (TMPnano) added to a 35% hydrogen peroxide (H2O2) bleaching gel on the color alteration, enamel mechanical and morphological properties, and H2O2 transamelodentinal diffusion. MATERIALS AND METHODS: Bovine enamel/dentin discs (n = 180) were divided according to the bleaching gel: 35% H2O2 (HP); 35% H2O2 + 0.1% F (HP/F); 35% H2O2 + 1% TMPnano (HP/TMPnano); 35% H2O2 + 0.1% F + 1% TMPnano (HP/F/TMPnano) and 35% H2O2 + 2% calcium gluconate (HP/Ca). The gels were applied 3 times by 40 min; once each 7-day. The Commission Internationale de l'Eclairage (CIE) L*a*b* total color alteration (ΔE), color alteration by CIEDE2000 (ΔE00), whitening index (ΔWID), surface (SH) and cross-sectional hardness (ΔKHN), surface roughness (Ra), and transamelodentinal diffusion were determined. Enamel surfaces were evaluated by Scanning Electron Microscopy (SEM) and X-ray Dispersive Energy (EDX). Data were submitted to ANOVA, followed by the Student-Newman-Keuls test (p <0.05). RESULTS: ΔE, ΔE00, and ΔWID were similar among the gels that promoted a bleaching effect after treatment (p <0.001). Mineral loss (SH and ΔKHN), Ra, and H2O2 diffusion were lower for HP/F/TMPnano; the HP and HP/Ca groups presented the highest values (p <0.001). For SEM/EDX, surface changes were observed in all bleached groups, but less intense with TMPnano. CONCLUSIONS: Gels containing F/TMPnano do not interfere with the bleaching effect and reduce enamel demineralization, roughness, H2O2 diffusion, and morphological changes. CLINICAL RELEVANCE: Whitening gels containing F/TMPnano can be used as a new strategy to increase safety and maintain clinical performance.

Effect of the association of microparticles and nano-sized ß-calcium glycerophosphate in conventional toothpaste on enamel remineralization: In situ study.

OBJECTIVES: This in situ study aimed to assess the remineralizing effect of a fluoride toothpaste supplemented with ß-calcium glycerophosphate in both micro (ß-CaGPm) and nano-sized forms (ß-CaGPn). METHODS: This blind and cross-over study was performed in 4 phases, each spanning 3 days. Twelve volunteers utilized palatal appliances containing four bovine enamel blocks with artificial caries lesions. Volunteers were randomly assigned to the following treatment groups: Placebo (no F-ß-CaGPm-ß-CaGPn); 1100 ppm F alone (1100F); 1100F plus 0.5% micrometric ß-CaGP (1100F-0.5%ß-CaGPm); and 1100F plus 0.25%nano-sized ß-CaGP (1100F-0.25%ß-CaGPn). Participants were instructed to brush their natural teeth with the palatal appliances in the mouth for 1 min (3 times/day), ensuring that the enamel blocks were exposed to the natural toothpaste slurries. Following each phase, evaluations were conducted to determine the percentage of surface hardness recovery (%SHR), integrated recovery of subsurface hardness (ΔIHR), profile subsurface lesion through polarized light microscopy (PLM), as well as fluoride (F), calcium (Ca), and phosphorus (P) concentrations within the enamel. Data were analyzed by ANOVA and Student-Newman-Keuls test (p < 0.001). RESULTS: Treatment with 1100F-0.25%ß-CaGPn resulted in %SHR ∼69 % and ∼40 % higher when compared to 1100F and 1100F-0.5%ß-CaGPm (p < 0.001). The reduction in lesion body (ΔIHR; PLM) was ∼40 % higher with 1100F-0.25%ß-CaGPn (p < 0.001) compared to 1100F. The addition of ß-CaGPm and ß-CaGPn did not influence enamel F concentration (p > 0.001). Treatment with 1100F-0.25%ß-CaGPn led to an increase in the concentration of Ca and P in the enamel (p < 0.001). CONCLUSION: The addition of 0.25%ß-CaGPn into 1100F formulation increased the bioavailability of calcium and phosphate, promoting a higher remineralizing effect. CLINICAL SIGNIFICANCE: Toothpaste containing 1100F-0.25%ß-CaGPn showed a potential of higher remineralization to 1100 ppm F and 1100 ppm F micrometric ß-CaGP could be a strategy for patients at caries activity.

Effect of fluoride gels with nano-sized sodium trimetaphosphate on the in vitro remineralization of caries lesions.

OBJECTIVE: To evaluate the effects of fluoride (F) gels supplemented with micrometric or nano-sized sodium trimetaphosphate (TMPmicro and TMPnano, respectively) on the in vitro remineralization of caries-like lesions. METHODOLOGY: Bovine enamel subsurface lesions (n=168) were selected according to their surface hardness (SH) and randomly divided into seven groups (n=24/group): Placebo (without F/TMP), 4,500 ppm F (4500F), 4500F + 2.5% TMPnano (2.5% Nano), 4500F + 5% TMPnano (5% Nano), 4500F + 5% TMPmicro (5% Micro), 9,000 ppm F (9000F), and 12,300 ppm F (Acid gel). The gels were applied in a thin layer for one minute. Half of the blocks were subjected to pH cycling for six days, whereas the remaining specimens were used for loosely- (calcium fluoride; CaF2) and firmly-bound (fluorapatite; FA) fluoride analysis. The percentage of surface hardness recovery (%SHR), area of subsurface lesion (ΔKHN), CaF2, FA, calcium (Ca), and phosphorus (P) on/in enamel were determined. Data (log10-transformed) were subjected to ANOVA and the Student-Newman-Keuls' test (p<0.05). RESULTS: We observed a dose-response relation between F concentrations in the gels without TMP for %SHR and ΔKHN. The 2.5% Nano and 5% Micro reached similar %SHR when compared with 9000F and Acid gels. For ΔKHN, Placebo and 5% Nano gels had the highest values, and 5% Micro, 2.5% Nano, 9000F, and Acid gels, the lowest. All groups had similar retained CaF2 values, except for Placebo and Acid gel. We verified observed an increase in Ca concentrations in nano-sized TMP groups. Regarding P, TMP groups showed similar formation and retention to 9000F and Acid. CONCLUSION: Adding 2.5% nano-sized or 5% micrometric TMP to low-fluoride gels lead to enhanced in vitro remineralization of artificial caries lesions.

In vitro effect of low-fluoride toothpaste supplemented with sodium trimetaphosphate, xylitol, and erythritol on enamel demineralization.

OBJECTIVE: Regular use of toothpaste with fluoride (F) concentrations of ≥ 1000 ppm has been shown to contribute to reducing caries increment. However, when used by children during the period of dental development, it can lead to dental fluorosis. In this study, we aimed to evaluate the in vitro effect of a toothpaste formulation with reduced fluoride (F) concentration (200 ppm) supplemented with sodium trimetaphosphate (TMP: 0.2%), Xylitol (X:16%), and Erythritol (E: 4%) on dental enamel demineralization. METHODOLOGY: Bovine enamel blocks were selected according to initial surface hardness (SHi) and then divided into seven experimental toothpaste groups (n=12). These groups included 1) no F-TMP-X-E (Placebo); 2) 16% Xylitol and 4% Erythritol (X-E); 3) 16% Xylitol, 4% Erythritol and 0.2%TMP (X-E-TMP); 4) 200 ppm F (no X-E-TMP: (200F)); 5) 200 ppm F and 0.2% TMP (200F-TMP); 200 ppm F, 16% Xylitol, 4% Erythritol, and 0.2% TMP (200F-X-E-TMP); and 7) 1,100 ppm F (1100F). Blocks were individually treated 2×/day with slurries of toothpastes and subjected to a pH cycling regimen for five days (DES: 6 hours and RE: 18 hours). Then, the percentage of surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN), fluoride (F), calcium (Ca), and phosphorus (P) in enamel were determined. The data were analyzed by ANOVA (1-criterion) and the Student-Newman-Keuls test (p<0.001). RESULTS: We found that the 200F-X-E-TMP treatment reduced %SH by 43% compared to the 1100F treatments (p<0.001). The ΔKHN was ~ 65% higher with 200F-X-E-TMP compared to 1100F (p<0.001). The highest concentration of F in enamel was observed on the 1100F treatment (p<0.001). The 200F-X-E-TMP treatment promote higher increase of Ca and P concentration in the enamel (p<0.001). CONCLUSION: The association of 200F-X-E-TMP led to a significant increase of the protective effect on enamel demineralization compared to the 1100F toothpaste.

Establishment of a saliva donor selection for in vitro biofilm growth / Seleção de doadores de saliva para crescimento de biofilme in vitro

Introdução: O emprego de biofilmes polimicrobianos, utilizando a saliva como inóculo, é um modelo promissor para o estudo de biofilmes cariogênicos in vitro. Entretanto, ainda não existe uma padronização para seleção de doadores de saliva. Objetivo: O objetivo deste estudo foi estabelecer uma metodologia para seleção de doadores de saliva utilizando fatores salivares microbianos e características in vitro do biofilme. Material e método: Para doação de saliva foram selecionados vinte voluntários. Os voluntários permaneceram 24 horas sem escovar os dentes e ficaram em jejum por 2 horas antes da coleta da saliva. Foram avaliados os seguintes parâmetros: viabilidade das bactérias anaeróbias totais e mutans streptococci; concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM) da clorexidina; capacidade de formação de biofilme por meio da biomassa; e a suscetibilidade dos biofilmes à clorexidina. Resultado: A viabilidade bacteriana da saliva, a capacidade de formação de biofilme e a suscetibilidade do biofilme à clorexidina foram apresentadas como média e intervalo de confiança (95%). A diferença entre a viabilidade do biofilme (mutans streptococci e bactérias totais) após tratamento com NaCl 0,9% e diacetato de clorexidina 0,2% foi comparada pelo teste t de Student com nível de significância estabelecido em 5%. A viabilidade total de bactérias anaeróbias (mediana) foi de 7,28 log 1+UFC/mL (unidades formadoras de colônia/mL). A viabilidade dos mutans streptococci na saliva apresentou mediana de 5,47 log 1+UFC/mL. Para capacidade de formação de biofilme a mediana da biomassa foi de 0,1172 A570. Conclusão: O tratamento com clorexidina reduziu significativamente os mutans streptococci e a viabilidade total das bactérias. A metodologia para seleção do doador de saliva foi estabelecida com sucesso.

Introduction: The utilization of polymicrobial biofilms, with saliva as an inoculum, represents a promising model for in vitro studies on cariogenic biofilms. However, there is still no standardization for selecting saliva donors. Objective: The aim of this study is to establish a methodology for the selection of saliva donors using microbial salivary factors and in vitro biofilm characteristics. Material and method: For saliva donation, twenty volunteers were selected. Volunteers remained 24 h without brushing their teeth and fasted for 2 h before saliva collection. The following parameters were evaluated: total anaerobic bacteria and mutans streptococci viability; minimum inhibitory concentration (MIC) and minimum bactericide concentration (MBC) of chlorhexidine; biofilm forming capacity by biomass assessment; and the susceptibility of biofilms to chlorhexidine. Result: Saliva bacterial viability, biofilm forming capacity and biofilm susceptibility to chlorhexidine were presented as mean and confidence interval (95%). The difference between biofilm (mutans streptococci and Total bacteria) viability after treatment with NaCl 0.9% and 0.2% chlorhexidine diacetate was compared using the Student t-test with a significance level established at 5%. Total anaerobic bacteria viability (median) was 7.28 log 1+CFU/mL (colony forming units/ mL). Mutans streptococci viability in the saliva showed a median of 5.47 log 1+CFU/mL. Biofilm forming capacity showed that biomass had a median of 0.1172 A570. Conclusion: Treatment with chlorhexidine significantly reduced mutans streptococci and total bacteria viability. The methodology for the selection of the saliva donor was successfully established.

In vitro effect of low-fluoride toothpaste supplemented with sodium trimetaphosphate, xylitol, and erythritol on enamel demineralization

Abstract Regular use of toothpaste with fluoride (F) concentrations of ≥ 1000 ppm has been shown to contribute to reducing caries increment. However, when used by children during the period of dental development, it can lead to dental fluorosis. Objective: In this study, we aimed to evaluate the in vitro effect of a toothpaste formulation with reduced fluoride (F) concentration (200 ppm) supplemented with sodium trimetaphosphate (TMP: 0.2%), Xylitol (X:16%), and Erythritol (E: 4%) on dental enamel demineralization. Methodology: Bovine enamel blocks were selected according to initial surface hardness (SHi) and then divided into seven experimental toothpaste groups (n=12). These groups included 1) no F-TMP-X-E (Placebo); 2) 16% Xylitol and 4% Erythritol (X-E); 3) 16% Xylitol, 4% Erythritol and 0.2%TMP (X-E-TMP); 4) 200 ppm F (no X-E-TMP: (200F)); 5) 200 ppm F and 0.2% TMP (200F-TMP); 200 ppm F, 16% Xylitol, 4% Erythritol, and 0.2% TMP (200F-X-E-TMP); and 7) 1,100 ppm F (1100F). Blocks were individually treated 2×/day with slurries of toothpastes and subjected to a pH cycling regimen for five days (DES: 6 hours and RE: 18 hours). Then, the percentage of surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN), fluoride (F), calcium (Ca), and phosphorus (P) in enamel were determined. The data were analyzed by ANOVA (1-criterion) and the Student-Newman-Keuls test (p<0.001). Results: We found that the 200F-X-E-TMP treatment reduced %SH by 43% compared to the 1100F treatments (p<0.001). The ΔKHN was ~ 65% higher with 200F-X-E-TMP compared to 1100F (p<0.001). The highest concentration of F in enamel was observed on the 1100F treatment (p<0.001). The 200F-X-E-TMP treatment promote higher increase of Ca and P concentration in the enamel (p<0.001). Conclusion: The association of 200F-X-E-TMP led to a significant increase of the protective effect on enamel demineralization compared to the 1100F toothpaste.

Effect of fluoride gels with nano-sized sodium trimetaphosphate on the in vitro remineralization of caries lesions

Abstract Objective To evaluate the effects of fluoride (F) gels supplemented with micrometric or nano-sized sodium trimetaphosphate (TMPmicro and TMPnano, respectively) on the in vitro remineralization of caries-like lesions. Methodology Bovine enamel subsurface lesions (n=168) were selected according to their surface hardness (SH) and randomly divided into seven groups (n=24/group): Placebo (without F/TMP), 4,500 ppm F (4500F), 4500F + 2.5% TMPnano (2.5% Nano), 4500F + 5% TMPnano (5% Nano), 4500F + 5% TMPmicro (5% Micro), 9,000 ppm F (9000F), and 12,300 ppm F (Acid gel). The gels were applied in a thin layer for one minute. Half of the blocks were subjected to pH cycling for six days, whereas the remaining specimens were used for loosely- (calcium fluoride; CaF2) and firmly-bound (fluorapatite; FA) fluoride analysis. The percentage of surface hardness recovery (%SHR), area of subsurface lesion (ΔKHN), CaF2, FA, calcium (Ca), and phosphorus (P) on/in enamel were determined. Data (log10-transformed) were subjected to ANOVA and the Student-Newman-Keuls' test (p<0.05). Results We observed a dose-response relation between F concentrations in the gels without TMP for %SHR and ΔKHN. The 2.5% Nano and 5% Micro reached similar %SHR when compared with 9000F and Acid gels. For ΔKHN, Placebo and 5% Nano gels had the highest values, and 5% Micro, 2.5% Nano, 9000F, and Acid gels, the lowest. All groups had similar retained CaF2 values, except for Placebo and Acid gel. We verified observed an increase in Ca concentrations in nano-sized TMP groups. Regarding P, TMP groups showed similar formation and retention to 9000F and Acid. Conclusion Adding 2.5% nano-sized or 5% micrometric TMP to low-fluoride gels lead to enhanced in vitro remineralization of artificial caries lesions.

Effect of glaze and chlorhexidine on physical and mechanical properties of bis-acryl resin: An in situ study.

BACKGROUND: Temporary prosthesis protects the oral tissues, in addition to providing aesthetic look and masticatory function until a definitive prosthesis is manufactured. OBJECTIVES: To evaluate the effect of glaze and 0.12% chlorhexidine (CHX) on the physical and mechanical properties of bis-acryl, and to evaluate the antimicrobial efficacy of CHX. MATERIAL AND METHODS: Eighty specimens of bis-acryl resin were made. Over 40 of them the glaze was applied. One specimen with and 1 specimen without glaze were placed in niches of an appliance manufactured for each patient. Each of the 20 volunteers received 2 devices. Initially, the volunteers used one device and treated it with sucrose for 7 days (control), and later they used the other device and treated it with sucrose and CHX for 7 days (test). Color, microhardness, roughness, surface energy, and insoluble extracellular polysaccharides (EPS) tests were performed. All results were submitted to the Tukey's test, with the exception of the EPS results, which were submitted to the Student's t test. RESULTS: The ΔE00 of the unglazed control group was significantly higher than that of the unglazed test group. In all groups, a significant decrease in microhardness occurred over time. At both times, the glaze significantly increased the microhardness of the specimens (in all the glazed groups). At the final time, the test glaze group showed significantly higher microhardness compared with the control glaze group. Roughness in the groups without glaze increased significantly with CHX treatment over time. At both times, the glaze generated a significant reduction in roughness in the control and test groups. There was a significant reduction in surface energy over time in all groups. In most comparisons, the glazed groups showed significantly higher surface energy values compared with the unglazed control group. At the final time point, the unglazed test group showed a significantly higher surface energy value than the unglazed control group; and the glazed test group showed a significantly higher surface energy value compared with the glazed control group. The resins that received CHX had a significantly lower amount of biofilm. CONCLUSIONS: Color values were clinically acceptable in all tested groups. At both time points, the roughness values were clinically acceptable only in the glazed groups. Glaze increased the microhardness of the specimens. Microhardness and surface energy were reduced over time in all groups. Chlorhexidine can help prevent microhardness degradation. Glaze and CHX can increase surface energy. Chlorhexidine reduced the amount of bacterial biofilm.

Pomegranate Extract Potentiates the Anti-Demineralizing, Anti-Biofilm, and Anti-Inflammatory Actions of Non-Alcoholic Mouthwash When Associated with Sodium-Fluoride Trimetaphosphate.

This study investigated the anti-caries and anti-inflammatory effects of mouthwash formulations containing Punica granatum (pomegranate) peel extract (PPE), sodium-trimetaphosphate, and low concentrations of fluoride. PPE was characterized using high-performance liquid chromatography (ellagic acid and punicalagin). Total phenolics were quantified among formulations, and their stability was analyzed for 28 days. The formulation effects were evaluated as follows: (1) inorganic component concentration and reduced demineralization on bovine enamel blocks subjected to pH cycling; (2) anti-biofilm effect on dual-biofilms of Streptococcus mutans ATCC 25175 and Candida albicans ATCC 10231 treated for 1 and 10 min, respectively; and (3) cytotoxicity and production of inflammatory mediators (interleukin-6 and tumor necrosis factor-alpha). The formulation containing 3% PPE, 0.3% sodium-trimetaphosphate, and 225 ppm of fluoride resulted in a 34.5% surface hardness loss; a 13% (treated for 1 min) and 36% (treated for 10 min) biofilm reduction in S. mutans; a 26% (1 min) and 36% (10 min) biofilm reduction in C. albicans; absence of cytotoxicity; and anti-inflammatory activity confirmed by decreased interleukin-6 production in mouse macrophages. Thus, our results provide a promising prospect for the development of an alcohol-free commercial dental product with the health benefits of P. granatum that have been recognized for a millennium.

Silver nanoparticles associated with a polyphosphate and fluoride enhance the prevention of enamel demineralization and impact on dual-biofilm adhesion.

OBJECTIVES: The aim of this study were to produce a multifunctional nanocomposite combining silver nanoaparticles (Ag), sodium trimetaphosphate (TMP) and fluoride (F), to investigate its effect on dental enamel demineralization and on biofilms of Streptococcus mutans and Candida albicans. METHODS: Bovine enamel blocks were submitted to five pH cycles and treated 2x/day with 100 ppm F, 225 ppm F, 100 ppm F + 0.2%TMP or 100 ppm F + 0.2%TMP+10% Ag (100F/TMP/Ag). Next, surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN), enamel fluoride (F) and calcium (Ca) concentration were determined. Biofilms from single and dual species of S. mutans and C. albicans were treated with 100F/TMP/Ag, Ag or chlorhexidine gluconate for 24 h. The antibiofilm effect was evaluated by colony-forming unit counting and Scanning Electron Microscopy. RESULTS: The nanocomposite reduced 43.0% of %SH and was similar with samples treated with 225F, 100F/TMP and 100/TMP/Ag. The attribute of F and/or TMP in reducing ΔKHN in 5-20 µm was not affected by the addiction of Ag (110F = 225F = 100F/TMP = 100F/TMP/Ag > Negative Control). Further, 100F/TMP/Ag strongly reduced viable cells of S. mutans in dual biofilms (∼5 log10cm2) and structurally affected the biofilms. CONCLUSION: The 100F/TMP/F promoted a protective effect against enamel demineralization and was able to significantly inhibit the growth of biofilms of S. mutans and C. albicans. CLINICAL SIGNIFICANCE: The focus on prevention and non-invasive dental treatment is the most effective and least costly way to improve the population's oral health conditions. We present a nanocomposite for a multiple approach in prevention of caries.

Fluoride and trimetaphosphate association as a novel approach for remineralization and antiproteolytic activity in dentin tissue.

OBJECTIVE: The study evaluated the effect of solutions containing fluoride (F) and/or sodium trimetaphosphate (TMP) and F/TMP on the inhibition of MMP-2 and MMP-9, and on dentin remineralization in vitro. DESIGN: Bovine root dentin blocks were prepared, and caries-like lesions were induced in two thirds of the surface. Blocks were then randomly divided into 13 groups/solutions (n = 10): Placebo; 0.3 %, 1 % and 3 % NaOH-hydrolyzed TMP; 0.3 %, 1 % and 3 % TMP; 250, 500 and 1100 ppm F; 250 ppm F + 0.3 % TMP; 500 ppm F + 1 % TMP and 1100 ppm F + 3 % TMP. One third of each specimen was treated with the respective solutions in pH-cycling. The mineral concentration (gHAp × cm-3 × µm) was determined by computed X-ray microtomography, and data submitted to ANOVA and Student-Newman-Keuls' test (p < 0.05). The ability of the solutions to inhibit MMP-2 and MMP-9 activity was assessed by zymography. RESULTS: F/TMP association led to less mineral loss in the deeper region of the lesion and reduced the depth of lesions when compared to its counterpart without TMP (p < 0.001). 3 % TMP (hydrolyzed or not), 500 ppm F and 1100 ppm F completely inhibited MMP-2 activity, while for MMP-9 such effects were only achieved by treatment with 1100 ppm F + 3 % TMP. CONCLUSION: Treatment with 1100 ppm F + 3 % TMP fully inhibits the gelatinolytic activity of MMPs-2 and - 9 and shows greater remineralizing capacity in artificial caries lesions in dentin. However, hydrolyzing TMP does not improve its anti-proteolytic activity and its remineralizing capacity.

Evaluation of bleaching efficacy, microhardness, and trans-amelodentinal diffusion of a novel bleaching agent for an in-office technique containing hexametaphosphate and fluoride.

OBJECTIVE: This study evaluated in vitro the effects of calcium gluconate (CaGlu), sodium fluoride (NaF), sodium hexametaphosphate (HMP), and NaF/TMP added to a 35% hydrogen peroxide (H2O2) bleaching gel on the color change, enamel hardness, and trans-amelodentinal diffusion. MATERIALS AND METHODS: Enamel discs/bovine dentin (n = 150) were divided according to the bleaching gel: 35% H2O2 (H2O2); 35% H2O2 + 0.1% NaF (H2O2/NaF); 35% H2O2 + 1% HMP (H2O2/HMP); 35% H2O2 + 0.1% NaF + 1% HMP (H2O2/NaF/HMP), and 35% H2O2 + 2% CaGlu (H2O2/Caglu). The bleaching gels were applied three times (40 min/session) at 7-day intervals between each application. Then, color alteration (ΔE), whitening index (ΔWID), percentage of surface hardness loss (% SH), cross-sectional hardness (ΔKHN), and trans-amelodentinal diffusion were determined. Data were submitted for analysis of variance (ANOVA), followed by the Student-Newman-Keuls test (p < 0.05). RESULTS: All bleaching gels showed significant color changes after treatment (p < 0.001). ΔE and ΔWID were similar among the evaluated gels. Mineral loss (% SH and ΔKHN) and trans-amelodentinal diffusion of hydrogen peroxide were lower for H2O2/NaF/HMP; the H2O2/CaGlu group presented the highest values about the other groups (p < 0.001). CONCLUSION: It is possible to conclude that the addition of NaF/HMP to the in-office bleaching agent did not interfere with the bleaching efficacy and reduced enamel demineralization and H2O2 diffusion. CLINICAL SIGNIFICANCE: The association of NaF/HMP to the bleaching gel can be used as a novel approach for minimizing the adverse effects of H2O2 by-products and with similar clinical efficacy.

Surface Free Energy, Interaction, and Adsorption of Calcium and Phosphate to Enamel Treated with Trimetaphosphate and Glycerophosphate.

This study aimed to evaluate the surface (γs) and interaction (ΔGiwi) free energy and calcium (Ca2+) and phosphate (PO43-) adsorption to dental enamel treated with sodium trimetaphosphate (TMP) or calcium glycerophosphate (CaGP) that had or had not been exposed to CaPO4-containing solutions. Bovine enamel blocks (n = 192; 24 blocks/group) were treated (2 mL/block; 2 min) with TMP (0%, 1%, 3%, and 9%) and CaGP (0, 0.25, 0.5, and 1%) or exposed to a CaPO4-containing solution. The adsorption of these compounds by enamel was assessed before and after treatment. γs and ΔGiwi and their apolar (γsLW and ΔGiwiLW) and polar (γsAB and ΔGiwiAB) components and acid-base interactions (γs+/γs-) were determined by the contact angles. The data were subjected to ANOVA, followed by the Student-Newman-Keuls test (p < 0.05). The adsorption of TMP was dose dependent (p < 0.001), and it reduced γs and γsAB and increased ΔGiwiAB (ΔGiwi > 0) and γs- when compared with the group without TMP (p < 0.001). The immersion in CaPO4-containing solution increased γs and γsAB and reduced ΔGiwiAB (ΔGiwi > 0) and γs- (p < 0.001). There was a correlation between the adsorption of TMP and Ca2+ (r = 0.916; p < 0.001) and PO43- (r = 0.899; p < 0.001). The adsorption of CaGP on the enamel was dose dependent (p < 0.001), reducing γs, ΔGiwiAB (ΔGiwi < 0), γsLW, and γs- when compared to the group without CaGP (p < 0.001). When exposed to the CaPO4-containing solution, there was an increase in ΔGiwiAB (ΔGiwi > 0), γsLW, and γs- and a decrease in γsAB (p < 0.001) without adsorption of Ca2+ by enamel. It may be concluded that TMP and CaGP were adsorbed onto the enamel, producing hydrophilic and hydrophobic surfaces, respectively. TMP produces electron donor sites that induce Ca2+ adsorption, while CaGP releases Ca2+ into the medium.