Antifungal activity of tyrosol and farnesol used in combination against Candida species in the planktonic state or forming biofilms.

AIM: This study examined the antifungal activity of the combination of tyrosol and farnesol against Candida albicans and Candida glabrata in the planktonic state or forming biofilms. METHODS AND RESULTS: The effect of drug association against Candida planktonic cells was assessed by the fractional inhibitory concentration index. Mono- and dual-species biofilms were developed during 24 h and then treated with the compounds for 3 days, with two daily treatments of 1 min each. After, the total biomass, metabolic activity and the number of cultivable cells were quantified. Planktonic cells of the two species showed a similar susceptibility to the drug combination, however, a synergistic effect was only verified for C. glabrata. Regarding biofilm susceptibility, significant reductions in C. glabrata biomass, metabolism of C. albicans and mixed biofilms, and cultivable cells of single biofilms were verified for the drug combination, indicating an additive effect. For all other experiments, the effects were classified as indifferent. CONCLUSION: The combined use of tyrosol and farnesol was advantageous for some of the analysed parameters against Candida species. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings may contribute to the development of oral care products containing tyrosol and farnesol to combat oral infections caused by Candida species.

Activity of tyrosol against single and mixed-species oral biofilms.

AIM: This study aimed to evaluate the effect of tyrosol on the formation of single and mixed biofilms of Candida albicans ATCC 10231, Candida glabrata ATCC 90030 and Streptococcus mutans ATCC 25175 formed on acrylic resin (AR) and hydroxyapatite (HA) surfaces. METHODS AND RESULTS: Single and mixed biofilms were formed on AR and HA in the presence of tyrosol at 50, 100 and 200 mmol l(-1), during 48 h. Next, antimicrobial activity was assessed through metabolic activity (XTT reduction assay) and the number of colony-forming units (CFUs). Scanning electron microscopy observations were performed in order to analyse biofilm structure. Tyrosol, mainly at 200 mmol l(-1), significantly decreased the metabolic activity and number of CFUs for all single and mixed-species biofilms formed on both surfaces. SEM images suggested cell damage caused by tyrosol. CONCLUSION: Tyrosol showed inhibitory effects against biofilms formed by important oral pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study showing the antibiofilm effect of tyrosol on Candida species and Strep. mutans in single and mixed cultures. These results may be useful in the development of topical therapies focused on preventing biofilm-associated oral diseases, such as denture stomatitis and dental caries.

In situ remineralizing effect of fluoride varnishes containing sodium trimetaphosphate.

OBJECTIVE: This study analyzed the effects of a fluoride (F) varnish supplemented with sodium trimetaphosphate (TMP) on the remineralization of caries-like lesions in situ. MATERIALS AND METHODS: Twelve subjects used palatal devices with demineralized enamel discs for 3 days, following a double-blind, crossover protocol. Test groups included placebo (no F or TMP), 5% NaF and 5% NaF/5% TMP varnishes. The percentage of surface hardness recovery (%SHR) and cross-sectional hardness (ΔKHN) were determined. RESULTS: Significant differences were observed among all varnishes regarding %SHR and ΔKHN. The highest %SHR and the lowest ΔKHN were seen for the 5% NaF/5% TMP varnish, followed by 5% NaF and placebo. CONCLUSION: The remineralizing effect of a 5% NaF varnish is significantly enhanced when associated with TMP. CLINICAL RELEVANCE: The reduction in the subsurface lesion area of enamel treated with the TMP-containing varnish implies that cavities would take longer to develop or might not develop at all depending on individual factors, resulting in lower net caries increments at individual and population levels.

In vitro remineralizing effect of fluoride varnishes containing sodium trimetaphosphate.

This study analyzed the effects of fluoride varnishes supplemented with sodium trimetaphosphate (TMP) on the remineralization of caries-like lesions in vitro. Bovine enamel discs were selected through surface hardness (SH) and caries-like lesions were induced. SH was again determined and the blocks were divided into 7 experimental groups (n = 24/group): placebo (no fluoride or TMP), 5% TMP, 2.5% NaF, 2.5% NaF/5% TMP, 5% NaF, 5% NaF/5% TMP and commercial formulation (Duraphat™, 5% NaF), following a blind protocol. Discs were treated with the varnishes and kept in a remineralizing solution for 4 h and a demineralizing solution for 2 h. Varnishes were then removed and half of the discs were used for analysis of loosely (CaF2) and firmly bound fluoride. The remaining discs were submitted to a pH-cycling regimen for 6 days. The percentage of surface hardness recovery (%SHR), cross-sectional hardness (ΔKHN) and enamel CaF2 and fluoride were determined. Data were analyzed by ANOVA and by Student-Newman-Keuls' test (p < 0.05). A dose-response relationship was observed between fluoride concentrations in the varnishes and %SHR. The 5% TMP varnish led to %SHR similar to that obtained for the placebo. When TMP was used in association with fluoride, however, significantly higher %SHR were observed in comparison with their counterparts without TMP. Moreover, ΔKHN obtained for the 5% NaF/5% TMP was significantly lower among all groups tested. Higher concentrations of CaF2 and fluoride were observed for Duraphat and 5% NaF, followed by 5% NaF/5% TMP, 2.5% NaF and 2.5% NaF/5% TMP (p < 0.05). It was concluded that the supplementation of fluoride varnishes with TMP leads to enhanced remineralizing effect of artificial caries lesions in vitro.

In vitro evaluation of the effect of mouth rinse with trimetaphosphate on enamel demineralization.

OBJECTIVE: The aim of this study was to investigate the effectiveness of sodium trimetaphosphate (TMP) addition to mouth rinses to inhibit enamel demineralization. DESIGN: Bovine enamel blocks (n = 88) were selected by surface hardness and divided into eight treatment groups (n = 11 per group): placebo, 100 or 225 µg F/ml; the rinses with 100 µg F/ml had differing TMP concentrations (range 0-0.6%). The blocks were subjected to pH cycling for 5 days and treated twice a day with mouth rinses. After that, surface and cross-sectional hardness as well as fluoride in enamel were measured. RESULTS: The groups containing both 100 µg F/ml and 0.4% TMP inhibited demineralization most effectively (p < 0.001). This formulation yielded lower values of lesion areas than the formulations containing 100 or 225 µg F/ml but no TMP. The addition of 0.4% TMP increased the fluoride in enamel. CONCLUSION: It is possible to improve the effectiveness of a mouth rinse with 100 µg F/ml by addition of TMP, this being superior in inhibiting enamel demineralization compared with mouth rinses containing 225 µg F/ml.

Effect of iron II on hydroxyapatite dissolution and precipitation in vitro.

The aim of this study was to evaluate the effect of iron II on the dissolution and precipitation of synthetic hydroxyapatite (HA). HA powder was suspended in solutions of iron (0.84 µg/ml, Fe0.84; 18.0 µg/ml, Fe18; 70.0 µg/ml, Fe70), fluoride (1,100 µg/ml, F1,100), and deionized water and submitted to pH cycling. After pH cycling, the samples were analyzed by infrared spectroscopy and X-ray diffraction. The concentrations of fluoride, calcium, phosphorus, and iron were also analyzed. The data were submitted to ANOVA, and analyzed by Tukey's test (p < 0.05). The infrared spectrum showed a reduction in all bands corresponding to phosphates and hydroxyls and an increase in the carbonate band in the groups with iron. The intensity of the phosphate bands increased and that of the hydroxyl bands decreased in the group F1,100. It was observed that there was a higher concentration of Ca in the group F1,100, with no significant difference between the groups Fe18 and Fe70 (p > 0.05). There was an increase in Fe concentration in the HA directly related to the Fe concentration of the treatment solutions. Results show that the presence of Fe causes the precipitation of apatite with high solubility.

Evaluation of different fluoride concentrations supplemented with trimetaphosphate on enamel de- and remineralization in vitro.

Trimetaphosphate (TMP) effects on demineralized bovine enamel were studied after 15 days of pH cycling. Treatments included 30 wt% (weight percent) dilutions of 0, 500, 1,500 or 3,000 µg F/g aqueous NaF solutions with or without 3% TMP. Treated specimens were assessed by transverse microradiography. With the exception of the 3,000 µg F/g case, 3% TMP addition provided significant additional overall remineralization compared with F alone. Mineral content profiles differed significantly between corresponding F and F + TMP groups. Fluoride alone resulted in more remineralization in the original demineralized zone, whereas F + TMP caused less demineralization in the underlying, originally sound enamel.

Effect of mouth rinses with fluoride and trimetaphosphate on enamel erosion: an in vitro study.

The effect of mouth rinses containing fluoride (100 µg/ml) and sodium trimetaphosphate (TMP) on enamel erosion was evaluated in vitro. Bovine enamel blocks were subjected to erosive challenges 4 times per day for 5 min, followed by treatment with placebo, 225 µg F/ml, 100 µg F/ml, 100 µg F/ml and TMP (0.2, 0.4 and 0.6%) solutions (30 s) and storage in artificial saliva, over a duration of 5 days. TMP groups showed lower enamel wear than fluoride-only and placebo groups (p < 0.05). Addition of TMP at a TMP:NaF molar proportion between 1.24:1 and 3.72:1 to a solution containing 100 µg F/ml presented a greater protective effect under erosive conditions than a solution containing 225 µg F/ml, in the absence of TMP.

Use of fluoridated dentifrices by children in Araçatuba, Brazil: factors affecting brushing habits and amount applied on the brush.

PURPOSE: To evaluate the type and the amount of fluoridated dentifrice applied on children's toothbrushes by parents/guardians according to descriptions typically recommended by scientific societies, as well as to assess the influence of demographic and socioeconomic variables on dentifrice use. METHODS: Parents/guardians of children (0-7 years old; n = 306; convenience sample) attending vaccination centres from Araçatuba (Brazil), answered to a structured questionnaire comprising items related to interviewees' education, child's age, gender, brushing habits and use of fluoridated dentifrice. The amount of toothpaste used by children during toothbrushing was estimated using a portable scale. Similarly, the interviewees were requested to apply dentifrices on toothbrushes according to eight descriptions, ranging from "smear" to "all bristles", following a random sequence. Data were submitted to Mann-Whitney's, Kruskal-Wallis' and Friedman's tests, and Spearman's correlation coefficient (p < 0.05). RESULTS: The type of toothpaste and the amount of product used at home were not affected by the respondents' educational level or family income. However, child's age was significantly correlated with the amount of toothpaste placed on the toothbrush (r = 0.324, p < 0.001). Also, the amount of toothpaste placed on the toothbrush increased according to what would be expected from the descriptions, although wide variations were observed within each description, with large interquartile and overall ranges. CONCLUSION: The amount and the type of dentifrice used by children were influenced by their age, while parents/caregivers' interpretation on verbal instructions regarding appropriate dentifrice quantities varied widely. This reinforces the need for educative measures on the appropriate use by dentifrices by children.

Low-fluoride acidic dentifrice: a randomized clinical trial in a fluoridated area.

BACKGROUND: Low-fluoride dentifrices have been suggested as alternatives to reduce dental fluorosis risk, but there is no consensus regarding their clinical effectiveness, which has been suggested to be increased when their pH is acidic. AIMS: This single-blind randomized clinical trial evaluated the caries increment during the use of a low-fluoride acidic liquid dentifrice. METHODS: Four-year-old schoolchildren (n = 1,402) living in a fluoridated area (0.6-0.8 ppm F) were randomly allocated to 4 groups differing according to the type of dentifrice used over a 20-month period. Group 1 (n = 345): liquid dentifrice, 1,100 ppm F, pH 4.5. Group 2 (n = 343): liquid dentifrice, 1,100 ppm F, pH 7.0. Group 3 (n = 354): liquid dentifrice, 550 ppm F, pH 4.5. Group 4 (n = 360): toothpaste, 1,100 ppm F, pH 7.0. At baseline and after 20 months, clinical examinations were conducted (dmfs index) and caries increment was calculated. Data were analysed by GLM procedure using classrooms (cluster) as unit of analysis (p < 0.05). RESULTS: The mean ± SD (95% CI) net increments found were as follows. Group 1: 2.06 ± 2.38 (1.8-2.3); group 2: 2.08 ± 2.87 (1.7-2.4); group 3: 2.05 ± 2.79 (1.7-2.4), and group 4: 2.08 ± 2.34 (1.8-2.4). No significant differences were detected among the groups. CONCLUSION: In a population with high caries risk living in a fluoridated area, as the selected sample, and according to the present protocol, the low-fluoride acidic liquid dentifrice seems to lead to similar caries progression rates as conventional 1,100 ppm F toothpaste.

Effect of different fluoride concentrations of experimental dentifrices on enamel erosion and abrasion.

It has been suggested that fluoride products are able to reduce erosive tooth wear. Thus, the purpose of this in vitro study was to evaluate the effect of dentifrices with different fluoride concentrations as well as of a low-fluoridated dentifrice supplemented with trimetaphosphate (TMP) on enamel erosion and abrasion. One hundred twenty bovine enamel blocks were assigned to the following experimental dentifrices: placebo, 1,100 microg F/g, 500 microg F/g plus 3% TMP and 5,000 microg F/g. The groups of enamel blocks were additionally subdivided into conditions of erosion (ERO) and of erosion plus abrasion (ERO + ABR). For 7 days, the blocks were subjected to erosive challenges (immersion in Sprite 4 times a day for 5 min each time) followed by a remineralizing period (immersion in artificial saliva between erosive challenges for 2 h). After each erosive challenge, the blocks were exposed to slurries of the dentifrices (10 ml/sample for 15 s). Sixty of the blocks were additionally abraded by brushing using an electric toothbrush (15 s). The alterations of the enamel were quantified using the Knoop hardness test and profilometry (measurements in micrometers). The data were analyzed using a 2-way ANOVA test followed by a Bonferroni correction (p < 0.05). In in vitro conditions, the 5,000 microg F/g and 500 microg F/g plus 3% TMP dentifrices had a greater protective effect when compared with the 1,100 microg F/g dentifrice, under both ERO and ERO + ABR conditions. The results suggest that dentifrices alone are not capable of completely inhibiting tooth wear.

Cross-sectional microhardness of human enamel subjected to erosive, cariogenic or combined erosive/cariogenic challenges.

This in situ study evaluated the interaction between caries and erosion processes. In the first phase, enamel specimens were subjected to erosion without dental plaque (EO) or to erosion with plaque (EP); in the second phase, they were subjected to erosion plus cariogenic challenge (EC) or cariogenic challenge (CO), both with plaque accumulation. Cross-sectional hardness data (10-330 microm depth) were tested using ANOVA (alpha = 0.05). EO and EP showed surface softening to 10 microm depth. CO and EC produced subsurface lesions, of similar depth (up to 220 microm), with CO showing higher integrated loss of hardness than EC, indicating that cariogenic and erosive challenges did not have an additive effect.

In vitro evaluation of dentifrice with low fluoride content supplemented with trimetaphosphate.

One way to reduce dental fluorosis is by reducing the fluoride (F) concentration in dentifrice, but low-F dentifrice should be as effective as a standard dentifrice. This study evaluated in vitro whether the supplementation with sodium trimetaphosphate (TMP) of a dentifrice with low F content (500 microg/g) would provide a similar effect to that of a standard dentifrice. Bovine enamel blocks were submitted to a pH cycling regime incorporating daily exposures to a slurry of dentifrice: a low-F dentifrice with or without 0.1-3.0% TMP; an F-free, phosphate-free dentifrice (negative control), or a dentifrice with 1,100 microg/g F (positive control). The addition of TMP to dentifrice with or without F was associated with higher surface hardness and decreased loss of integrated subsurface hardness after pH cycling. The combination of 1% TMP and 500 microg F/g had a greater effect than the positive control dentifrice. It is concluded that the addition of TMP to the 500-microg F/g dentifrice allowed a similar or larger effect as compared with a standard dentifrice in this in vitro model.

Comparison of methods for evaluating mineral loss: hardness versus synchrotron microcomputed tomography.

This study analyzed degrees of demineralization in bovine enamel using synchrotron microcomputed tomography (SMCT) and hardness measurements (Knoop hardness number, KHN). For 5 days, 40 bovine enamel blocks were individually subjected to a pH cycling model and treatment with fluoride dentifrices (placebo, 275, 550 and 1,100 microg F/g) diluted in deionized water twice a day. Surface hardness number and cross-sectional profiles of hardness and mineral concentration (by SMCT) were determined. Integrated hardness (KHN x microm) for sound and demineralized specimens was calculated and subtracted to give the integrated loss of hardness (DeltaKHN) for the lesions. Increasing fluoride concentration in the dentifrices led to higher values for surface hardness after pH cycling and mineral concentration (g(HAp) cm(-3)), and lower values for DeltaKHN (p < 0.05). From the present results, it may be concluded that hardness measurements revealed demineralization in all groups, which was lower in groups treated with dentifrice with a higher F concentration. SMCT and hardness measurements gave similar results in areas with higher demineralization, but diverged in areas with lower demineralization.

Evaluation of laser fluorescence in the monitoring of the initial stage of the de-/remineralization process: an in vitro and in situ study.

This study aimed to evaluate laser fluorescence (LF) for monitoring the initial stage of subsurface de- and remineralization (<150 microm depth). Ninety-six sound blocks of bovine enamel, selected according to surface hardness (SH) and LF were used in two experimental studies, in vitro and in situ. In vitro, blocks were exposed to a demineralizing solution, then remineralized by pH cycling for 6 days. In situ, 10 volunteers wore acrylic palatal appliances, each containing 4 dental enamel blocks that were demineralized for 14 days by exposure to 20% sucrose solution. Following this treatment, blocks were submitted to remineralization for 1 week with fluoride dentifrice (1,100 microg F/g). In both experiments, SH and LH were measured after demineralization and after remineralization. Further, enamel blocks were selected after the demineralization/remineralization steps for measurement of cross-sectional hardness and integrated loss of subsurface hardness (Delta KHN). SH and Delta KHN showed significant differences among the phases in each study. LF values for sound, demineralized and remineralized enamel were: 5.2 +/- 1.1, 8.1 +/- 1.2 and 5.6 +/- 0.8, respectively, in the in vitro study, and 5.3 +/- 0.3, 16.5 +/- 4.7 and 6.5 +/- 2.5, respectively, in the in situ study, values for demineralized enamel being significantly higher than for sound and remineralized enamel in both studies. However, LF was correlated with Delta KHN only in situ. LF was capable of monitoring de- and remineralization in early lesions in situ, when bacteria are presumably present in the caries lesion body, but is not correlated with mineral changes in bacteria-free systems.

Synchrotron microComputed Tomography of the mature bovine dentinoenamel junction.

The mature dentinoenamel junction (DEJ) is viewed by some investigators and the current authors, not as a fossilized, sharp transition between enamel and dentin, but as a relatively broad structural transition zone including the mantle dentin and the inner aprismatic enamel. In this study, the DEJ structure in bovine incisors was studied with synchrotron microComputed Tomography (microCT) using small cubes cut parallel to the tooth surface. The reconstructions revealed a zone of highly variable punctate contrast between bulk dentin and enamel; the mean linear attenuation coefficients and their standard deviations demonstrated that this zone averaged less mineral than dentin or enamel but had more highly variable structure than either. The region with the punctuate contrast is, therefore, the mantle dentin. The thickness of the mantle dentin seen in a typical data set was about 30 microm, and the mantle dentin-enamel interface deviated +/-15 microm from the average plane over a distance of 520 microm. In the highest resolution data ( approximately 1.5 microm isotropic voxels, volume elements), tubules in the dentin could be discerned in the vicinity of the DEJ. Contrast sensitivity was high enough to detect differences in mineral content between near-surface and near-DEJ volumes of the enamel. Reconstructions before and after two cubes were compressed to failure revealed cracks formed only in the enamel and did not propagate across the mantle dentin, regardless of whether loading was parallel to or perpendicular to the DEJ.

Effect of Psidium cattleianum leaf extract on Streptococcus mutans viability, protein expression and acid production.

Plants naturally produce secondary metabolites that can be used as antimicrobials. The aim of this study was to assess the effects of Psidium cattleianum leaf extract on Streptococcus mutans. The extract (100%) was obtained by decoction of 100 g of leaves in 600 ml of deionized water. To assess killing, S. mutans biofilms were treated with water (negative control) or various extract dilutions [100, 50, 25% (v/v) in water] for 5 or 60 min. To evaluate the effect on protein expression, biofilms were exposed to water or 1.6% (v/v) extract for 120 min, proteins were extracted and submitted to 2-dimensional difference gel electrophoresis. Differentially expressed proteins were identified by mass spectrometry. The effect of 1.6% (v/v) extract on acid production was determined by pH measurements and compared to a water control. Viability was similar after 5 min of treatment with the 100% extract or 60 min with the 50% extract (about 0.03% survival). There were no differences in viability between the biofilms exposed to the 25 or 50% extract after 60 min of treatment (about 0.02% survival). Treatment with the 1.6% extract significantly changed protein expression. The abundance of 24 spots was decreased compared to water (p < 0.05). The extract significantly inhibited acid production (p < 0.05). It is concluded that P. cattleianum leaf extract kills S. mutans grown in biofilms when applied at high concentrations. At low concentrations it inhibits S. mutans acid production and reduces the expression of proteins involved in general metabolism, glycolysis and lactic acid production.

Effect of a 4% titanium tetrafluoride (TiF4) varnish on demineralisation and remineralisation of bovine enamel in vitro.

OBJECTIVES: This in vitro study assessed the effect of a 4% TiF4 varnish on demineralisation and remineralisation of sound enamel and artificial carious enamel lesions, respectively. METHODS: Bovine sound and carious enamel (n=110) were randomly allocated to each type of varnish: Duraphat-D (NaF, 2.26%F, pH 4.5, Colgate-Brazil, n=30), Duofluorid-F (NaF, 2.71%F, pH 8.0, FGM-Brazil, n=30), TiF4-T (2.45%F, pH 1.0, FGM-Brazil, n=30) and no-fluoride-P (FGM-Brazil, pH 5.0, n=20). For the formation of artificial enamel caries, half of the blocks were immersed in 32 mL buffer acetate solution (16 h), whereas the other half was maintained sound. The varnishes were applied onto the enamel surfaces. Thus, the samples were subjected to pH cycles (37 degrees C) for 7 days. The response variables tested were surface and cross-sectional hardness. Data were tested using Kruskal-Wallis test (p<0.05). RESULTS: All F varnishes significantly reduced demineralisation and increased remineralisation in comparison to placebo. The TiF4 did not significantly reduce the surface enamel softening when compared with the other F varnishes, but it decreased the loss of subsurface hardness to the same extent. In enamel blocks with previous artificial carious lesions, the TiF4 significantly improved the rehardening compared to the other varnishes up to 30 microm depth. CONCLUSIONS: The TiF4 varnish was able to decrease the demineralisation and increase the remineralisation of previously sound and carious enamel, respectively. It was equally effective compared to NaF varnishes on reducing the demineralisation at subsurface, but it was more effective on improving the remineralisation at surface and subsurface.

In vitro evaluation of the effectiveness of acidic fluoride dentifrices.

This study evaluated the effectiveness of acidic low-fluoride dentifrices compared to conventional neutral dentifrices. Enamel blocks were submitted to pH cycling and treatment with slurries of dentifrices containing 0, 275, 412, 550 and 1,100 microg F/g (pH 4.5 or 7.0), and also a commercial dentifrice (1,100 microg F/g) and a commercial children's dentifrice (500 mug F/g). Variations in surface microhardness and in the mineral content in enamel after pH cycling were calculated. Enamel blocks treated with acidic dentifrices exhibited less mineral loss compared to neutral dentifrices (ANOVA; p < 0.05). The acidic dentifrices with 412 and 550 microg F/g had the same effectiveness as the neutral 1,100-microg F/g dentifrice and commercial 1,100-microg F/g dentifrice.

Toothpaste with Nanosized Trimetaphosphate Reduces Enamel Demineralization.

This double-blind crossover study assessed the effects of a low-fluoride (low-F) dentifrice containing nanosized sodium trimetaphosphate (TMP) on enamel demineralization in situ. Nineteen subjects wore palatal appliances containing 4 blocks of bovine enamel and were randomly assigned to brush their teeth with placebo (without F/TMP), 250-ppm F (250F), 250F plus 0.05% nanosized TMP (250F-TMPnano), and 1,100-ppm F (1,100F) dentifrices during 7 d, under cariogenic challenge. Enamel surface hardness and cross-sectional hardness (ΔKHN [Knoop hardness number]), as well as F, calcium (Ca), and phosphorus (P) concentrations, were determined. Also, biofilm that formed on the blocks was analyzed for F, Ca, P, and insoluble extracellular polysaccharide concentrations. Data were submitted to analysis-of-variance models and Student-Newman-Keuls test ( P < 0.05). The 250F-TMPnano dentifrice promoted the lowest ΔKHN among all groups ( P < 0.001), while the percentage of surface hardness loss was similar to 1,100F. Also, similar F, Ca, and P concentrations in enamel were observed for 1,100F and 250F-TMPnano. In the biofilm, the highest F content was observed for 1,100F; Ca content was similar between 1,100F and 250F-TMPnano; and P content was similar among all groups. Similar extracellular polysaccharide values were observed for 250F-TMPnano and 1,100F ( P < 0.001), ionic activity of CaHPO40, CaF+, and HF0 ( P < 0.05) and degree of saturation of hydroxyapatite and CaF2 ( P < 0.05). It was concluded that the protective effect of 250F-TMPnano dentifrice was similar to a conventional dentifrice for most of the variables studied, having a more pronounced effect on the subsurface lesion when compared with the conventional toothpaste (1,100F). Knowledge Transfer Statement: Although toothpastes containing ≥1,000-ppm fluoride are more effective than low-fluoride formulations against dental caries, their early use can lead to side effects. This has prompted intensive research on alternatives to increase the anticaries effect of low-fluoride toothpastes. The present in situ study demonstrated that the addition of sodium trimetaphosphate nanoparticles to toothpastes containing 250-ppm fluoride significantly enhances the protective effect of this formulation against enamel demineralization to levels comparable to a 1,100-ppm fluoride toothpaste in terms of most of the variables studied. Most important, this formulation promoted the lowest loss of subsurface hardness among all groups, suggesting that caries lesions would take longer to develop under clinical conditions when compared with a conventional (1,100-fluoride) toothpaste.