Effectiveness of Fluoride Varnishes for White Spot Lesion Prevention and Remineralization during Orthodontic Treatment: A Randomized Controlled Trial.

INTRODUCTION: This study investigated the effectiveness of titanium tetrafluoride (TiF4) varnish compared to sodium fluoride (NaF) varnish for preventing and remineralizing white spot lesions (WSLs) in individuals undergoing orthodontic treatment. METHODS: This randomized, placebo-controlled, and double-blinded study was conducted with sixty-five adolescents who were selected based on caries activity and then randomized into three parallel groups: G1 (placebo varnish), G2 (5%-NaF varnish) and G3 (4%-TiF4 varnish). Volunteers received varnish application weekly for the first 4 weeks, after 6 (T1) and 12 (T2) months. The measured outcomes included: prevention of new WSLs, and reversal/progression of WSLs, assessed by Nyvad and ICDAS indices, as well as quantitative light-induced fluorescence (QLF). Chi-square, ANOVA, and Kruskall-Wallis tests were applied. The level of significance was set at 0.05, and post hoc Bonferroni test for p values were performed to correct for multiple comparisons. RESULTS: 1,274 teeth were included; 70.5% were Nyvad 0, and 29.5% were Nyvad 1, with no differences between the groups at baseline (T0). Regarding ICDAS, 70.5% were ICDAS 0, 21.6% were ICDAS 1, and 7.9% were ICDAS 2. G1 showed an increasing prevalence of WSLs at T1 and maintained stable at T2; G2 exhibited a decline at T2, while G3 experienced a decrease at T1 and T2 (p < 0.01). Incidence of WSLs at T2 was 10.2% (G1), 5.6% (G2) and 1.4% (G3). The percentage of teeth initially scored as Nyvad 0 that progressed to Nyvad 1 was 13%, 6.8%, and 1% for G1, G2, and G3, respectively. Conversely, the percentage of teeth initially scored as Nyvad 1 that regressed to Nyvad 0 or Nyvad 2 (T0-T2) was 14%, 49.3%, and 74.4%, respectively (p < 0.001). As for ICDAS index, regression was observed in 6.5%, 17.8% and 24%, while progression was observed in 14.9%, 7.7% and 0.9% for G1, G2 and G3, respectively (p < 0.001). Significant differences among the 3 groups for integrated fluorescence loss (mean±SD, G1: -14.28±9.47, G2: -11.10±11.49 and G3: -6.77±11.00) were found at T2 (p < 0.01). CONCLUSION: Both varnishes demonstrated the ability to prevent and remineralize WSLs. However, TiF4 varnish exhibited the most effective control over WSLs during the 12-month orthodontic treatment.

A sugarcane cystatin (CaneCPI-5) alters microcosm biofilm formation and reduces dental caries.

The antimicrobial and anticaries effects of CaneCPI-5 were evaluated. Ninety bovine enamel samples were treated for 60 s with either phosphate-buffered-saline (PBS), 0.12% chlorhexidine (CHX), 0.05 mg ml-1 CaneCPI-5, 0.1 mg ml-1 CaneCPI-5 or 0.5 mg ml-1 CaneCPI-5. They were incubated with inoculum (human saliva + McBain's saliva) for the first 8 h. From then until the end of the experiment, the enamel was exposed to McBain saliva with sucrose and, once a day, for 5 days, they were treated with the solutions. At the end of the experimental period, resazurin and viable plate count assays were performed. Enamel demineralization was also measured. All concentrations of CaneCPI-5 and CHX significantly reduced the activity of biofilms compared with PBS. For viable plate counts, all treatments similarly reduced the lactobacilli and total streptococci; for the mutans streptococci, 0.05 mg ml-1 CaneCPI-5 performed better than CHX. All CaneCPI-5 concentrations significantly reduced the integrated mineral loss. This study represents the first step regarding the use of CaneCPI-5 within the concept of acquired enamel pellicle and biofilm engineering to prevent dental caries.

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.

Influence of irradiated dentin, biofilm and different artificial saliva formulations on root dentin demineralization.

The aim of this study was evaluated the influence of radiation as well as of new formulations of artificial saliva on the development of root dentin lesions. Bovine root samples were divided into: irradiated (70 Gy) dentin or not; the type of biofilm (from irradiated patient-experimental or non-irradiated patient-control) and the type of artificial saliva (for the condition irradiated dentin/biofilm from irradiated patient): Control Artificial Saliva (inorganic); Control Saliva + 1 mg/ml hemoglobin; Control Saliva +0.1 mg/ml cystatin; Control Saliva + hemoglobin + cystatin; Bioextra (positive control) and deionized water (DiW, negative control) (n = 12/group). Biofilm was produced using human biofilm and McBain saliva (0.2 % of sucrose, 37o C and 5 % CO2); the treatments were done 1x/day, for 5 days. Colony-forming units (CFU) counting was performed; demineralization was quantified by transversal microradiography. Two-way ANOVA/Bonferroni or Sidak test for the comparison between biofilm x dentin and ANOVA/Tukey or Kruskal-Wallis/Dunn for comparing artificial saliva were done (p < 0.05). The type of biofilm had no influence on CFU and demineralization. Sound dentin under control biofilm presented the lowest Lactobacillus ssp. and Streptococcus mutans CFU and the lowest mean mineral loss (R) (25.6 ± 2.2; 23.7 ± 2.9 %) compared to irradiated dentin (26.1 ± 2.8; 28.1 ± 3.3, p < 0.004) for both types of biofilms (experimental and control, respectively). Bioextra was the only artificial saliva that reduced R (10.8 ± 2.5 %) and Lesion Depth (LD) (35 ± 15 µm) compared to DiW (17.3 ± 3.3 %, 81 ± 18 µm, p < 0.0001). Irradiation has impact on caries development; the experimental saliva were unable to reduce its occurrence.

Acquired pellicle and biofilm engineering with CaneCPI-5: Insights from proteomic and microbiomics analysis.

OBJECTIVE: In this in vivo proof-of-concept study, acquired pellicle engineering was implemented to promote alterations in the protein composition of the acquired enamel pellicle (AEP) and the bacterial composition of the dental biofilm after treatment with Sugarcane cystatin (CaneCPI-5). DESIGN: After prophylaxis, 10 volunteers rinsed (10 mL, 1 min) with the following solutions: 1) deionized water (H2O- negative control or 2) 0.1 mg/mL CaneCPI-5. The AEP and biofilm were formed along 2 or 3 h, respectively. The AEP was collected with electrode filter papers soaked in 3 % citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. The biofilm microbiome was collected with a dental curette. The DNA was extracted, amplified, and analyzed by 16S-rRNA Next Generation Sequencing (NGS). RESULTS: Treatment with CaneCPI-5 increased several proteins with antimicrobial, acid-resistance, affinity for hydroxyapatite, structural and calcium binding properties, such as Cysteine-rich-3 (6-fold-p = 0.03), Cystatin-B (5.5-fold-p < 0.01), Neutrophil-defensin 1 (4.7-fold-p < 0.01), Mucin (3.9-fold-p < 0.01), Immunoglobulin-heavy-constant (3.8-fold-p < 0.01) and Lactotransferrin (2.8-fold-p < 0.01). Microbiome revealed that several commensal bacteria had their abundance increased after rinsing with CaneCPI-5, such as Corynebacterium and Neisseria, while Streptococcus and Prevotella nigrescens were decreased. The results indicate the efficiency of CaneCPI-5 in promoting beneficial changes in the AEP and biofilm, making this phytocystatin a potential target for incorporation into dental products. CONCLUSION: Cane demonstrated the capability to alter the protein composition of the acquired enamel pellicle (AEP) and the initial colonizers of the biofilm, enhancing the presence of proteins and bacteria crucial for dental protection.

The protective effect of the experimental TiF4 and chitosan toothpaste on erosive tooth wear in vitro.

This study evaluated the protective effect of TiF4 and chitosan toothpaste on erosive tooth wear (ETW) in vitro. Enamel and dentin samples were randomly assigned to toothpastes (n = 12): (G1) TiF4 (1400 ppm F-), (G2) 0.5% chitosan (75% deacetylation, 500 mPas), (G3) TiF4 (1400 ppm F-) plus 0.5% chitosan (75% deacetylation, 500 mPas), (G4) Placebo, (G5) Erosion Protection (Elmex-GABA, 1400 ppm F-). Twelve samples were only eroded. All samples were submitted to erosive pH cycles and G1 to G5 to abrasive challenges using toothpastes' slurries plus 45 s of treatment, for 7 days. The final profile was overlaid to the baseline one for the ETW calculation (µm). The data were subjected to Kruskal-Wallis/Dunn tests. TiF4 toothpastes, regardless of the presence of chitosan, were able to significantly reduce ETW compared to placebo, while chitosan alone was similar to placebo for both tissues. The toothpastes containing TiF4 were even superior to the commercial Elmex toothpaste on enamel, while they were similar on dentin; both were also significantly different from placebo for both tissues. TiF4 and Elmex toothpastes minimized the impact of brushing on eroded surface. In conclusion, TiF4 toothpastes, regardless the presence of chitosan, showed to be effective in minimizing ETW in vitro.

Effect of TiF4/NaF and chitosan solutions on the development of enamel caries under a microcosm biofilm model.

OBJECTIVES: To evaluate the effect of experimental solutions containing TiF4/NaF and chitosan on bacterial species and on enamel caries prevention. METHODS: Microcosm biofilm was produced from human saliva mixed with McBain saliva (0.2% sucrose) on bovine enamel for five days, under 5% CO2 and 37 °C. From the second day until the end, the treatments were applied (1 × 60 s/day): (1) NaF (500 ppm F-, positive control); (2) TiF4 and NaF (TiF4: 190 ppm Ti4+ and 300 ppm F-; NaF: 190 ppm F-); (3) similar to 2 plus 0.5% chitosan (Ch 500 mPas, 75% deacetylation); (4) phosphate buffer solution (negative control); and (5) 0.5% chitosan (Ch 500 mPas, 75% deacetylation). CFU counting was performed for total microorganism, total streptococcus, total lactobacillus and Streptococcus mutans. Enamel demineralization was measured by transverse microradiography-TMR. The data were compared using ANOVA/Tukey or Kruskal-Wallis/Dunn tests (p < .050). RESULTS: No differences were found between the treatments with respect to CFU counting (ANOVA, p > .050). Enamel treated with TiF4/NaF plus chitosan solution presented the lowest demineralization compared to the negative control and pure chitosan solution. On the other hand, this experimental solution did not significantly differ from TiF4/NaF and NaF solutions, being all of them able to significantly reduce mineral loss (50-74%), but only TiF4/NaF plus chitosan reduced lesion depth (55%) compared to the negative control (p = .001). CONCLUSION: TiF4/NaF plus chitosan solution had no antimicrobial effect, but it was able to reduce enamel caries development in 79% compared to control under this model. CLINICAL SIGNIFICANCE: This study showed that TiF4/NaF plus chitosan solution had no antimicrobial effect, but it was able to reduce enamel caries development under a microcosm biofilm model.

Effect of sweetener containing Stevia on the development of dental caries in enamel and dentin under a microcosm biofilm model.

OBJECTIVE: This study compared the effect of commercial and pure sweetener containing stevia to that of aspartame, to sucrose and xylitol on the development of dental caries. METHODS: 228 bovine enamel and root dentin were exposed to microcosm biofilm model using human saliva. From the 2nd to the 5th day, the samples were exposed daily to McBain saliva supplemented with 0.2% of the respective sweeteners/sugar, under 5% CO2 and 37 °C. The lactic acid and the colony-forming units (CFU) were quantified. The demineralization was analyzed by TMR. The data were compared statistically (Kruskal-Wallis/ Dunn, p<0.05). RESULTS: Pure stevia, pure aspartame, xylitol and control were able to significantly reduce 92% of lactate production compared to sucrose. Stevia finn, aspartame finn and sucrose showed similar production of lactic acid (around 0.45±0.12 g/L and 0.67±0.18 g/L, for enamel and dentin, p<0.0001). With respect to total lactobacilli and S. mutans/S. sobrinus CFU, xylitol and control did not show growth on enamel, while CFU numbers were found in stevia finn, aspartame finn and sucrose groups for both tissues. Enamel and dentin demineralization was significantly reduced for xylitol, control, pure stevia and pure aspartame (85% and 83% reduction, respectively) compared to stevia finn, aspartame finn and sucrose, which in turn did not differ from each other (sucrose ΔZ: 2913.7 ± 646.7 vol%.µm for enamel and 3543.3 ± 432.5 vol%.µm for dentin). CONCLUSIONS: Commercial sweeteners containing stevia and aspartame proved to be as cariogenic as sucrose, which may be due to the other components, since the pure forms were not cariogenic. CLINICAL RELEVANCE: Our study showed that some commercial sweeteners (aspartame and stevia) are as cariogenic as sucrose, which may be due to the presence of lactose. The population should be advice about the presence of lactose in such brand names, to avoid their consume.

Effect of TiF4/NaF and chitosan solutions on biofilm formation and prevention of dentin demineralization.

OBJECTIVE: This study evaluated the effect of experimental solutions containing TiF4/NaF and chitosan on bacterial species of microcosm biofilm and on dentin demineralization. DESIGN: Microcosm biofilm was produced from human saliva mixed with McBain medium (0.2% sucrose) on bovine dentin for 5 days, under 5% CO2 and 37 °C. From the 2nd day to 5th day, the treatments were applied (1×60s/day) as following: (1) NaF (500 ppm F-, positive control); (2) TiF4 and NaF (TiF4: 190 ppm Ti4+ and 300 ppm F-; NaF: 190 ppm F-); (3) similar to 2 plus 0.5% chitosan (Ch 500 mPa.s, 75% deacetylation); (4) phosphate buffer solution (negative control); and (5) 0.5% chitosan (Ch 500 mPa.s, 75% deacetylation). CFU counting was performed for total microorganism, total streptococci, total lactobacilli and mutans streptococci. Dentin demineralization was measured by transverse microradiography-TMR. The data were compared using ANOVA/Tukey or Kruskal-Wallis/Dunn tests (p < 0.05). RESULTS: No differences were found between the treatments with respect to CFU counting (p > 0.05). Dentin treated with TiF4/NaF plus chitosan solution presented the lowest demineralization compared to the negative control and pure chitosan solution. On the other hand, this experimental solution did not significantly differ from TiF4/NaF solution, being both able to significantly reduce mineral loss. CONCLUSION: TiF4/NaF plus chitosan solution, at suitable pH to be clinically applicable, had no antimicrobial effect, but it was able to reduce dentin caries development under this model.

Acceptability and effect of TiF4 on dental caries: a randomized controlled clinical trial.

This randomized three-armed controlled clinical trial compared the effect of titanium tetrafluoride (TiF4) and sodium fluoride (NaF) varnishes on caries control in smooth surfaces of permanent dentition and children's acceptability. Sixty children (6-8 y/o) were randomly divided into TiF4 (2.45% F-), NaF (2.26% F-) or placebo (control) groups. Varnishes were applied on permanent teeth once a week for the first 4 weeks and after the 6th and 12th months of the study. The variables were as follows: International Caries Detection and Assessment System (ICDAS) scores, quantitative fluorescence changes, visual plaque index (VPI) and degree of acceptability. Two-way RM-ANOVA, ANOVA/Tukey and χ2 tests were performed (p < 0.05). No differences were found between the treatments with respect to ICDAS scores (p = 0.32). Only TiF4 reduced the mean fluorescence loss significantly at 18 months compared to the baseline (p = 0.003). TiF4 showed a lower percentage of new caries lesions by tooth surface than the placebo, while NaF did not induce such a change (p < 0.014). Regardless of the treatment, more than 95% of the participants reported being satisfied. For all groups, the VPI decreased significantly at 3 months compared to the baseline value (p < 0.001), with no differences between the treatments (p = 0.17). TiF4 had a similar ability to control caries lesions as NaF; however, only TiF4 differed from the placebo (p = 0.004). The acceptability of TiF4 varnish was similar to that of NaF varnish.

Protective effect of titanium tetrafluoride and silver diamine fluoride on radiation-induced dentin caries in vitro.

This in vitro study evaluated the protective effect of titanium tetrafluoride (TiF4) varnish and silver diamine fluoride (SDF) solution on the radiation-induced dentin caries. Bovine root dentin samples were irradiated (70 Gy) and treated as follows: (6 h): 4% TiF4 varnish; 5.42% NaF varnish; 30% SDF solution; placebo varnish; or untreated (negative control). Microcosm biofilm was produced from human dental biofilm (from patients with head-neck cancer) mixed with McBain saliva for the first 8 h. After 16 h and from day 2 to day 5, McBain saliva (0.2% sucrose) was replaced daily (37 °C, 5% CO2) (biological triplicate). Demineralization was quantified by transverse microradiography (TMR), while biofilm was analyzed by using viability, colony-forming units (CFU) counting and lactic acid production assays. The data were statistically analyzed by ANOVA (p < 0.05). TiF4 and SDF were able to reduce mineral loss compared to placebo and the negative control. TiF4 and SDF significantly reduced the biofilm viability compared to negative control. TiF4 significantly reduced the CFU count of total microorganism, while only SDF affected total streptococci and mutans streptococci counts. The varnishes induced a reduction in lactic acid production compared to the negative control. TiF4 and SDF may be good alternatives to control the development of radiation-induced dentin caries.

Effect of TiF4 varnish after pre-treatment with proanthocyanidin or chlorhexidine on the progression of erosive dentin loss in the presence or absence of the demineralized organic matrix.

This study evaluated the protective effect of TiF4 varnish, after pre-treatment with proantocyanidin or chlorhexidine, on the progression of erosive dentin loss (EDL), under the presence or absence of the demineralized organic matrix (DOM). Bovine root dentin samples were eroded for 30 min (0.1% citric acid, pH 2.5) and the loss was measured by profilometry. Half of them were subjected to the DOM removal using collagenase for 4 days, while the other half remained immersed in water. The removal of DOM was checked by profilometry. Samples were divided into 24 groups (n = 15) according to the factors: 1- With or without DOM; 2- Pre-treatment with 0.012% chlorhexidine gel, 10% proanthocyanidin gel or untreated for 1 min; 3-Final Treatment with TiF4 varnish, NaF varnish, placebo varnish or untreated for 6 h. The samples were submitted to a pH cycling for 5 days: 0.1% citric acid (4 × 90s/day) and artificial saliva between the challenges. The final profile was obtained for the calculation of EDL (µm, three-way ANOVA/Tukey test). When DOM was preserved, the EDL was lower compared to the condition without DOM (7.08 ± 4.03 µm and 9.80 ± 3.79 µm, p < 0.001, respectively), regardless of the treatments. The pre-treatment had no influence on the progression of EDL (p = 0.637), while the final treatment (TiF4 varnish only, 6.77 ± 4.08 µm) was effective in reducing the progression of EDL (NaF varnish: 9.52 ± 4.02 µm; Placebo varnish: 8.64 ± 4.06 µm and no treatment: 8.80 ± 3.95 µm). It can be concluded that DOM has important protective effect on the progression of EDL. TiF4 was the unique treatment capable of reducing EDL progression, regardless of the pre-treatment of DOM.

The effect of commercial whitening toothpastes on erosive dentin wear in vitro.

OBJECTIVES: This study evaluated the erosive tooth wear promoted by commercial whitening toothpastes on eroded dentin in vitro. DESIGN: Ninety bovine roots were embedded, polished and subjected to the baseline profile analysis. The samples were protected in 2/3 of the dentin surface and were randomly assigned to 6 groups (n = 15/group): Oral-B 3D White; Close-up Diamond Attraction Power White; Sorriso Xtreme White 4D; Colgate Luminous White; Crest and erosion only. All samples were submitted to erosive pH cycles (4 × 90 s in 0.1% citric acid, pH 2.5, per day) and abrasive challenges (2 × 15 s, per day) for 7 days. The samples were subjected to abrasion, using toothbrushing machine, soft toothbrushes and slurries of the tested toothpastes (1.5 N, 1:3 water). Between the challenges, the samples were immersed in artificial saliva. The final profile was overlaid to the baseline profile for the calculation of the erosive dentin wear (µm). The data were subjected to Kruskal-Wallis/Dunn tests (p < 0.05). RESULTS: Colgate Luminous White (4.7 µm) and Sorriso Xtreme White 4D (4.0 µm) promoted the highest wear, similarly to Oral-B 3D White (2.3 µm). Oral-B 3D White promoted similar wear compared to Crest (1.1 µm) and Close-up Diamond Attraction Power White (1.2 µm); however, it induced significant higher dentin wear compared to erosion only (1.0 µm). Close-up Diamond Attraction Power White and Crest did not increase the erosive wear compared to erosion only. CONCLUSION: Some whitening toothpastes increase the wear of eroded dentin, which should be considered by the dentist when prescribing them to patient with root exposure.

Effect of different citrus sweets on the development of enamel erosion in vitro.

The increased consumption of citrus sweets can contribute to the development of erosive tooth wear (ETW). Objective This in vitro study evaluated the erosive potential of citrus sweets on bovine enamel samples regarding the quantification of wear. Methodology Ninety bovine crowns were prepared and samples were randomly distributed into 6 groups (n=15): 0.1% citric acid solution (pH 2.5); Coca-Cola ® Soft Drink (pH 2.6); Fini ® Diet (lactic and citric acid, pH 3.3); Fini ® Jelly Kisses (lactic and citric acid, pH 3.5); Fini ® Fruit Salad Bubblegum (maleic acid, pH 2.6); Fini ® Regaliz Acid Tubes (maleic and citric acid, pH 3.1). Sweets were dissolved in the proportion of 40 g/250 mL of deionized water. Enamel samples were submitted to erosive challenges for 7 days (4 daily acid immersion cycles for 90 s each). Enamel wear was measured using contact profilometry (µm), and data (median values [interquartile range]) were submitted to Kruskal-Wallis/Dunn's test (p<0.0001). Results All citrus sweets tested present a high erosive potential, Fini Diet ® (2.4 [1.2]) and Fini Regaliz Tubs ® (2.2 [0.5]) show the highest erosive potential, similar to 0.1% citric acid (2.3 [0.7]); Fini Regaliz Tubs ® is more erosive than Coca-Cola ® (1.4 [0.9]). Conclusion The evaluated citrus sweets have great erosive potential and play a key role in the development of ETW.

Effect of titanium tetrafluoride/sodium fluoride solutions containing chitosan at different viscosities on the protection of enamel erosion in vitro.

OBJECTIVE: The aim of this in vitro study was to evaluate the protective effect against enamel erosion of experimental solutions containing TiF4/NaF and Chitosan compared to a commercial SnCl2/NaF/AmF solution. DESIGN: Bovine enamel samples were divided (n = 15/group) into: (1) commercial solution SnCl2/NaF/AmF (500 ppm F-, positive control); (2) NaF/TiF4 (490 ppm F-); (3) similar to 2 plus 0.5 % chitosan (Ch) (500 mPas), (4) similar to 2 plus 0.5 % chitosan (2000 mPas), (5) negative control (water), (6) 0.5 % chitosan (500 mPas) and (7) 0.5 % chitosan (2000 mPas). The samples were submitted to a pH cycling (0.1 % citric acid, 4 × 90 s/day, interposed by artificial saliva) and daily treatment application (after the last erosive challenge, 1 × 30 s/day) for seven days. After the first day, the surface reflection intensity changes (% rSRI) were measured. After 7 days, the erosive enamel loss was quantified by contact profilometer. The % rSRI and the enamel loss (µm) were compared using ANOVA/Tukey and Kruskal-Wallis/Dunn, respectively (p < 0.05). RESULTS: The solution containing TiF4/NaF plus Ch 500 mPas was the only able to reduce the early erosive demineralization compared to negative control (p = 0.003). However, it did not differ from the other solutions. Enamel samples treated with SnCl2/NaF/AmF presented the lowest median loss value [0.72 (0.18) µm] followed by both TiF4 + Ch [1.24 (0.49) and 1.28 (0.25)]; which significantly differed from the negative control [1.70 (0.27)]. CONCLUSION: The experimental solution containing TiF4/NaF plus chitosan (2000 mPas) has comparable effect to SnCl2/NaF/AmF on the protection against enamel erosion.

Antibiofilm and anti-caries effects of an experimental mouth rinse containing Matricaria chamomilla L. extract under microcosm biofilm on enamel.

OBJECTIVE: This study evaluated the antibiofilm and anti-caries effects of an experimental mouth rinse containing aqueous extract of Matricaria chamomilla L. METHODS: Microcosm biofilm was produced on bovine enamel, from pooled human saliva mixed with McBain saliva, under 0.2 % sucrose exposure, for 5 days. The biofilm was daily treated using (1 mL/1 min): Vochysia tucanorum Mart. (2.5 mg/mL); Myrcia bella Cambess. (1.25 mg/mL); Matricaria chamomilla L. (20 mg/mL); Malva sylvestris (Malvatricin® Plus-Daudt); 0.12 % Chlorhexidine (PerioGard®-Palmolive, Positive control) and PBS (Negative control). The % dead bacteria, biofilm thickness, EPS biovolume, lactic acid concentration, the CFU counting (total microorganisms, Lactobacillus sp., total streptococci and Streptococcus mutans/S. sobrinus) were determined. Enamel demineralization was measured by TMR. RESULTS: All mouth rinses induced bacterial death compared to PBS (p < 0.0001). The biofilm thickness varied from 12 ±â€¯2 µm (chlorhexidine) to 18 ±â€¯2 µm (V. tucanorum) (ANOVA/Tukey, p < 0.0001). The EPS biovolume varied from 7(4)% (chlorhexidine) to 30(20)% (PBS) (Kruskal-Wallis/Dunn, p < 0.0001). The lactic acid production was reduced by M. sylvestris (1.1 ±â€¯0.2 g/L) and chlorhexidine (0.6 ±â€¯0.2 g/L) compared to PBS (2.6 ±â€¯1.3 g/L) (ANOVA, p < 0.0001). Malva sylvestris and chlorhexidine showed significant low CFU for total microorganisms, Lactobacillus sp. and total streptococci. Only chlorhexidine significantly reduced S. mutans/S. sobrinus. CFUs for total streptococci and Lactobacillus sp, were also significantly reduced by M. chamomilla L. Malva sylvestris (63.4 % of mineral loss reduction), chlorhexidine (47.4 %) and M. chamomilla L. (39.4 %) significantly reduced enamel demineralization compared to PBS (ANOVA/Tukey, p < 0.0001). CONCLUSION: M. chamomilla L. has lower antibiofilm action, but comparable anti-caries effect to those found for chlorhexidine, under this model. CLINICAL RELEVANCE: This study shows that the antibiofilm and anti-caries potential may vary between the commercial and experimental mouth rinses containing natural agents, with promising results for those containing Matricaria chamomilla L. and Malva Sylvestris.

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

Abstract 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.

Antimicrobial and Anti-Caries Effect of New Glass Ionomer Cement on Enamel Under Microcosm Biofilm Model.

The occurrence of caries lesions adjacent to restorations is a serious problem in Dentistry. Therefore, new antimicrobial restorative materials could help to prevent recurrent carious lesions. This study evaluated the effect of a new glass ionomer cement (Ion Z) on the viability of a microcosm biofilm and on the development of enamel demineralization. Enamel samples were filled with the following materials (n=9): A) Ion-Z (FGM Ltda); B) Maxxion R (FGM Ltda); C) Ketac Fil Plus (3M ESPE) and D) no restoration (control). The samples were then exposed to human saliva mixed with McBain saliva (1:50) containing 0.2% sucrose for 14 days. The live and dead bacteria were quantified by fluorescence using a confocal laser-scanning microscope. The enamel demineralization was analyzed using transverse microradiography (TMR). The data were submitted to ANOVA/Tukey or Kruskal-Wallis/Dunn test (p<0.05). Ion Z induced a higher percentage of dead bacteria (60.96±12.0%) compared to the other groups (Maxxion R: 39.8±6.7%, Ketac Fil Plus: 43.7±9.71% and control 46.3±9.5%). All materials significantly reduced the average mineral loss compared to control (Ion-Z 25.0±4.2%vol, Maxxion R 23.4±8.0%vol, Ketac Fil Plus 30.7±7.7 and control 41.2±6.6%vol). Ion-Z was the only material able to significantly improve the mineral content at the surface layer (Zmax: 63.5±18.2%vol) compared to control (38.9±11.3%vol). Ion-Z shows antimicrobial potential, but its anti-caries effect was similar to the other materials, under this model.

Effect of an experimental mouth rinse containing NaF and TiF4 on tooth erosion and abrasion in situ.

OBJECTIVE: This study compared the effect of an experimental NaF/TiF4 mouth rinse with a commercial tin/F mouth rinse on the prevention of tooth wear in situ. METHODS: Fifteen subjects took part in this crossover and double-blind study, in which they wore a palatal appliance with 8 bovine teeth samples (4 enamel and 4 root dentine) in each of 3 phases (5 days each). Half of the samples were subjected to erosive challenges, and the other half to erosive plus abrasive challenges. The phases corresponded to the use of 1) Experimental solution containing NaF/TiF4 (189 ppm Ti+4, 500 ppm F-, pH 4.4); 2) commercial solution containing SnCl2/NaF/AmF (800 ppm Sn+2, 500 ppm F-, pH 4.5, Elmex®/GABA, positive control); 3) distilled water (negative control). Erosive challenges were performed using 0.1% citric acid (pH 2.5) for 90 s 4 times per day. The abrasion was done using a toothbrush and slurry of fluoride toothpaste, for 15 s 2 times per day. Thereafter, the subjects rinsed with the tested mouth rinse for 60s. Tooth wear was measured using contact profilometry (µm) and submitted to a two-way RM ANOVA/Bonferroni test (p < 0.05). RESULTS: No significant differences were detected between the experimental and the commercial mouth rinses, regardless of the challenge. Both fluoride mouth rinses were able to significantly reduce tooth wear compared to the negative control (p < 0.0001). No significant differences were detected with respect to tooth wear between the challenges (erosion and erosion plus abrasion). CONCLUSION: The experimental NaF/TiF4 mouth rinse has a similar protective effect to the commercial one against tooth wear in situ. CLINICAL SIGNIFICANCE: The experimental NaF/TiF4 solution protected against tooth wear in situ, regardless of the challenge (erosion or erosion plus abrasion), for both enamel and dentine, similarly to a commercial solution (tin/F-Elmex®) applied for this proposal. This result supports the conduction of clinical trials and a possible application of this solution in the future.

Effect of a Titanium Tetrafluoride Varnish in the Prevention and Treatment of Carious Lesions in the Permanent Teeth of Children Living in a Fluoridated Region: Protocol for a Randomized Controlled Trial.

BACKGROUND: Titanium tetrafluoride (TiF4) has regained interest due to new formulations that have been shown to be more effective against tooth demineralization than sodium fluoride (NaF) formulations in vitro and in situ. OBJECTIVE: The aim of this study is to evaluate the effect of two types of varnishes (4% TiF4 and a commercial 5% NaF) on the prevention of carious lesions and the treatment of noncavitated enamel carious lesions in the permanent teeth of children living in a fluoridated area. METHODS: This randomized, controlled, parallel and single-blind clinical trial involves 63 children, 6-7 years old, living in Bauru, São Paulo, Brazil. Children were selected according to their caries activity (ie, presence of at least 1 tooth with a Nyvad score of 1) and randomly divided into the following treatment categories: 4% TiF4 varnish (2.45 % F-, pH 1, FGM); 5% NaF varnish (2.26% F-, pH 5, Duraphat, Colgate) and control (placebo varnish, pH 5, FGM). The varnishes will be applied on all permanent teeth, once a week for 4 weeks and they will be reapplied only once 6 and 12 months after the study begins. Two calibrated examiners will carry out the clinical examination (International Caries Detection and Assessment System [ICDAS] and Nyvad indexes, kappa>.8) at baseline, before the first application, after the 1st, 6th, 12th, and 18th month of the study begins. Furthermore, quantitative fluorescence changes will be measured using Quantitative Light-Induced Fluorescence (QLF). The degree of patient satisfaction with the treatment will also be computed. The data will undergo statistical analysis (P<.05). RESULTS: This ongoing study is funded by funding agencies from Brazil (São Paulo Research Foundation, FAPESP-015/14149-1, and National Council for Scientific and Technological Development, CNPq-401313/2016-6). We expect to confirm the efficacy of TiF4 on the prevention and treatment of carious lesions by comparing it to NaF varnish. The subjects are under 1 month evaluation and the dropout was about 8%. No differences between the treatments have been detected at the first month so far (P>.05). CONCLUSIONS: If our hypothesis is confirmed, TiF4 varnish can be marketed and applied at the individual level and used in community programs to control dental caries. TRIAL REGISTRATION: Brazilian Clinical Trials Registry: RBR-5VWJ4Y; http://www.ensaiosclinicos.gov.br/rg/?q=RBR-5VWJ4Y (Archived by WebCite at http://www.webcitation.org/6wUurEnm7).