Riboflavin Attenuates Fluoride-Induced Testicular Injury via Interleukin 17A-Mediated Classical Pyroptosis.

Male reproductive toxicity of fluoride is of great concern worldwide, yet the underlying mechanism is unclear. Pyroptosis is a novel mode of inflammatory cell death, and riboflavin with anti-inflammatory properties has the potential to protect against fluoride damage. However, it is unknown whether pyroptosis is involved in fluoride-induced testicular injury and riboflavin intervention. Here, we first found that riboflavin could alleviate fluoride-caused lower sperm quality and damaged testicular morphology by reducing pyroptosis based on a model of ICR mice treated with NaF (100 mg/L) and/or riboflavin supplementation (40 mg/L) via drinking water for 13 weeks. And then, together with the results of in vitro Leydig cell modelsm it was confirmed that the pyroptosis occurs predominantly through classical NLRP3/Caspase-1/GSDMD pathway. Furthermore, our results reveal that interleukin-17A mediates the process of pyroptosis in testes induced by fluoride and riboflavin attenuation according to the results of our established models of riboflavin- and/or fluoride-treated IL-17A knockout mice. The results not only declare a new mechanism by which fluoride induces testicular injury via interleukin 17A-mediated classical pyroptosis but also provide evidence for the potential clinical application of riboflavin as an effective therapy for fluoride toxicity.

Amelogenin-inspired peptide, calcium phosphate solution, fluoride and their synergistic effect on enamel biomimetic remineralization: an in vitro pH-cycling model.

BACKGROUND: Several methods were introduced for enamel biomimetic remineralization that utilize a biomimetic analogue to interact and absorb bioavailable calcium and phosphate ions and induce crystal nucleation on demineralized enamel. Amelogenin is the most predominant enamel matrix protein that is involved in enamel biomineralization. It plays a major role in developing the enamel's hierarchical microstructure. Therefore, this study was conducted to evaluate the ability of an amelogenin-inspired peptide to promote the remineralization potential of fluoride and a supersaturated calcium phosphate solution in treating artificially induced enamel carious lesions under pH-cycling regimen. METHODS: Fifty enamel slices were prepared with a window (4*4 mm2 ) on the surface. Five samples were set as control healthy enamel and 45 samples were subjected to demineralization for 3 days. Another 5 samples were set as control demineralized enamel and 40 enamel samples were assigned into 8 experimental groups (n=5) (P/I, P/II, P/III, P/AS, NP/I, NP/II, NP/III and NP/AS) according to peptide treatment (peptide P or non-peptide NP) and remineralizing solution used (I; calcium phosphate solution, II; calcium phosphate fluoride solution, III; fluoride solution and AS; artificial saliva). Samples were then subjected to demineralization/remineralization cycles for 9 days. Samples in all experimental groups were evaluated using Raman spectroscopy for mineral content recovery percentage, microhardness and nanoindentation as healthy, demineralized enamel and after pH-cycling. Data were statistically analysed using two-way repeated measures Anova followed by Bonferroni-corrected post hoc test for pairwise multiple comparisons between groups. Statistical significance was set at p= 0.05. Additionally, XRD, FESEM and EDXS were used for crystal orientation, surface morphology and elemental analysis after pH-cycling. RESULTS: Nanocrystals clumped in a directional manner were detected in peptide-treated groups. P/II showed the highest significant mean values in mineral content recovery (63.31%), microhardness (268.81±6.52 VHN), elastic modulus (88.74±2.71 GPa), nanohardness (3.08±0.59 GPa) and the best crystal orientation with I002/I300 (1.87±0.08). CONCLUSION: Despite pH changes, the tested peptide was capable of remineralizing enamel with ordered crystals. Moreover, the supplementary use of calcium phosphate fluoride solution with peptide granted an enhancement in enamel mechanical properties after remineralization.

An in vitro evaluation of the effects of fluoride, CPP-ACP, or resin infiltration on discoloration caused by pediatric supplements.

OBJECTIVES: White spot lesions are more susceptible to staining agents due to their porous structure. This study examines the impact of white spot lesion treatments on discoloration caused by pediatric supplements. METHOD AND MATERIALS: Three treatments (fluoride, casein phosphopeptide-amorphous calcium phosphate [CPP-ACP], resin infiltration), a control, and their respective syrup-based subgroups (iron and black elderberry syrups) were established, each with eight teeth. Artificial white spot lesions were induced, and weekly applications of fluoride varnish, daily applications of CPP-ACP paste, or a single resin infiltration procedure were performed on the white spot lesions within the treatment groups over 4 weeks. Simultaneously, samples were exposed daily to iron or black elderberry syrups. Spectrophotometer measurements were taken at baseline, after demineralization (T0), and after 1 (T1), 2 (T2), and 4 weeks (T4). ΔE00 values were calculated. Statistical analysis was conducted using a three-way mixed-design ANOVA, with the significance level set at P = .05. RESULTS: At T4, ΔE00 values from all groups exceeded the clinical acceptability limit of 1.8. At T2 and T4, the ΔE00 values obtained from the black elderberry syrup subgroups were significantly higher (P < .001). At T4, the highest ΔE00 values were seen in the CPP-ACP groups (P < .001). The lowest ΔE00 values at T2 and T4 were observed in the resin infiltration groups (P < .05). CONCLUSIONS: Supplements containing ferrous sulfate and black elderberry extract caused color changes in white spot lesions that exceeded the clinical acceptability limit. Resin infiltration of white spot lesions provides advantages over remineralization treatments, particularly in minimizing discoloration induced by pediatric supplements.

Calcium supplementation attenuates fluoride-induced bone injury via PINK1/Parkin-mediated mitophagy and mitochondrial apoptosis in mice.

Excessive consumption of fluoride can cause skeletal fluorosis. Mitophagy has been identified as a novel target for bone disorders. Meanwhile, calcium supplementation has shown great potential for mitigating fluoride-related bone damage. Hence, this study aimed to elucidate the association between mitophagy and skeletal fluorosis and the precise mechanisms through which calcium alleviates these injuries. A 100 mg/L sodium fluoride (NaF) exposure model in Parkin knockout (Parkin-/-) mice and a 100 mg/L NaF exposure mouse model with 1% calcium carbonate (CaCO3) intervention were established in the current study. Fluoride exposure caused the impairment of mitochondria and activation of PTEN-induced putative kinase1 (PINK1)/E3 ubiquitin ligase Park2 (Parkin)-mediated mitophagy and mitochondrial apoptosis in the bones, which were restored after blocking Parkin. Additionally, the intervention model showed fluoride-exposed mice exhibited abnormal bone trabecula and mechanical properties. Still, these bone injuries could be effectively attenuated by adding 1% calcium to their diet, which reversed fluoride-activated mitophagy and apoptosis. To summarize, fluoride can activate bone mitophagy through the PINK1/Parkin pathway and mitochondrial apoptosis. Parkin-/- and 1% calcium provide protection against fluoride-induced bone damage. Notably, this study provides theoretical bases for the prevention and therapy of animal and human health and safety caused by environmental fluoride contamination.

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.

Green Tea-Derived Catechins Suppress the Acid Productions of Streptococcus mutans and Enhance the Efficiency of Fluoride.

Green tea-derived catechins, which can be divided into galloylated (epicatechin gallate: ECG, epigallocatechin gallate: EGCG) and non-galloylated (catechin: C, epicatechin: EC, epigallocatechin: EGC) catechins, are considered to be the main contributors to the caries control potential of green tea. In this study, we intended to compare the antimicrobial effects of these representative green tea-derived catechins and their combined effects with fluoride on the acid production and aggregation of Streptococcus mutans. The effects of different catechins on the growth, aggregation and acid production of S. mutans, and the combined effect of catechins and potassium fluoride (2 mm at pH 7.0, 0.3 mm at pH 5.5) on S. mutans acid production were measured by anaerobic culture, turbidity changes due to aggregation, and pH-stat methods. Molecular docking simulations were also performed to investigate the interactions between catechins and membrane-embedded enzyme II complex (EIIC), a component of the phosphoenolpyruvate-dependent phosphotransferase system (sugar uptake-related enzyme). ECG or EGCG at 1 mg/mL significantly inhibited the growth of S. mutans, induced bacterial aggregation, and decreased glucose-induced acid production (p < 0.05). All catechins were able to bind to EIIC in silico, in the following order of affinity: EGCG, ECG, EGC, EC, and C. Furthermore, they enhanced the inhibitory effects of fluoride at pH 5.5 and significantly inhibited S. mutans acid production by 47.5-86.6% (p < 0.05). These results suggest that both galloylated and non-galloylated catechins exhibit antimicrobial activity, although the former type demonstrates stronger activity, and that the caries control effects of green tea may be due to the combined effects of multiple components, such as catechins and fluoride. The detailed mechanisms underlying these phenomena and the in vivo effect need to be explored further.

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.

Dual protective effect of the association of plant extracts and fluoride against dentine erosion: In the presence and absence of salivary pellicle.

OBJECTIVES: To verify the protective effect of plant extracts associated with fluoride against dental erosion of dentine, in the presence and absence of a salivary pellicle. METHODS: Dentine specimens (n = 270) were randomly distributed into 9 experimental groups (n = 30/group): GT (green tea extract); BE (blueberry extract); GSE (grape seed extract); NaF (sodium fluoride); GT+NaF (green tea extract and NaF); BE+NaF (blueberry extract and NaF); GSE+NaF (grape seed extract and NaF); negative control (deionized water); and a positive control (commercialized mouthrinse containing stannous and fluoride). Each group was further divided into two subgroups (n = 15), according to the presence (P) or absence (NP) of salivary pellicle. The specimens were submitted to 10 cycles: 30 min incubation in human saliva (P) or only in humid chamber (NP), 2 min immersion in experimental solutions, 60 min of incubation in saliva (P) or not (NP), and 1 min erosive challenge. Dentine surface loss (dSL-10 and dSL-total), amount of degraded collagen (dColl) and total calcium release (CaR) were evaluated. Data were analyzed with Kruskal-Wallis, Dunn's and Mann-Whitney U tests (p>0.05). RESULTS: Overall, the negative control presented the highest values of dSL, dColl and CaR, and the plant extracts showed different degrees of dentine protection. For the subgroup NP, GSE showed the best protection of the extracts, and the presence of fluoride generally further improved the protection for all extracts. For the subgroup P, only BE provided protection, while the presence of fluoride had no impact on dSL and dColl, but lowered CaR. The protection of the positive control was more evident on CaR than on dColl. CONCLUSION: We can conclude that the plant extracts showed a protective effect against dentine erosion, regardless of the presence of salivary pellicle, and that the fluoride seems to improve their protection.

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.

Effect of laser acupuncture on pain and density of bone in osteoporotic postmenopausal women: a randomized controlled trial.

OBJECTIVE: This study aimed to evaluate the value of laser acupuncture (LA) on forearm bone mineral density (BMD) and wrist pain in osteoporotic postmenopausal women. METHODS: Sixty-eight postmenopausal women diagnosed with osteoporosis were randomly allocated equally to one of two sets. The drug-only group received calcium and vitamin D 3 supplement containing fluoride daily for 12 weeks, whereas the drug/LA group received LA therapy for 20 minutes per session, three sessions weekly, in addition to the same supplementation. The primary outcome parameter was assessment of BMD of the nondominant arm. Other outcomes included wrist pain. RESULTS: There was a highly significant improvement in the T-score of forearm BMD in both groups (-2.844 ± 0.476 to -2.597 ± 0.478 and -2.944 ± 0.486 to -1.652 ± 0.728 in the drug-only and drug/LA groups, respectively; P < 0.0001) and visual analog scale score (7.50 ± 0.79 to 4.24 ± 1.07 and 7.24 ± 0.82 to 3.09 ± 0.75 in the drug-only and drug/LA group, respectively; P < 0.0001). The improvement of both BMD and pain score was significantly higher in the drug/LA group (-1.303 and 4.15) compared with the drug-only group (-0.247 and 3.26; P < 0.0001). CONCLUSIONS: LA in combination with calcium and vitamin D supplementation containing fluoride is an effective modality in improving forearm BMD and reducing pain in osteoporotic postmenopausal women.

Resin-modified glass ionomer enriched with BIOGLASS: Ion-release, bioactivity and antibacterial effect.

Developing dental materials for the prevention of remineralization or demineralization is important for high-risk caries patients. This study aimed to evaluate the physicochemical and microbiological effects of adding 45S5 bioglass to resin-modified glass ionomer cement (RMGIC). Samples belonged to the following groups: GIC: conventional glass ionomer cement (Vitro Fil), RMGIC: resin-modified GIC (Vitro Fil LC), and RMGIC/45S5: RMGIC with 10% (wt %) of 45S5. Changes in pH and release of fluoride, calcium, and phosphorus ions under acidic (pH 4) and neutral (pH 7) pH conditions were evaluated. Antibacterial activity was verified based on colony-forming units. Material sorption and solubility were analyzed after bacterial exposure. After 28 days, the bioactivity of the materials was evaluated using scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS). Analysis of variance, post hoc Scheffe, and Tukey (α = 0.05) tests were employed for statistical analysis. RMGIC/45S5 showed higher alkalization activity, calcium release at pH 4 and 7, and sorption than GIC and RMGIC (p < .05). Release of phosphorus and fluoride at pH 4 and 7 was higher for GIC than that for RMGIC and RMGIC/45S5 (p < .05). RMGIC/45S5 showed higher values than RMGIC (p < .05). However, antibacterial activity did not differ among the groups. Precipitates of calcium and phosphorus were visualized in RMGIC/45S5 samples via SEM/EDS. These results indicate that the RMGIC/45S5 promotes alkalization and increases the release of calcium, phosphorus, and fluoride ions, resulting in precipitate deposition rich in calcium and phosphorus, thereby being a promising option to improve the bioactivity of RMGIC.

Miracle Tree (Moringa oleifera) Attuned GFAP and Synaptophysin Levels, Oxidative Stress and Biomarkers in Cerebellar Fluorosis of Pregnant Rats.

<b>Background and Objective:</b> Cerebellar fluorosis is a health issue associated with excessive exposure to fluoride (F) either in direct or indirect ways as pesticides, drinking water and caries preventing prescriptions. It is characterized by elevation in oxidative stress, inflammation, demyelination and Purkinje cell loss. <i>Moringa oleifera</i> (M), is a widely cultivated plant used as a health-booster agent in modulating various disorders because of its high content of vitamins and minerals. The beneficial effect of moringa against fluoride-induced cerebellar toxicity in pregnant rats was investigated in this study. <b>Materials and Methods:</b> Twenty pregnant rats were administered daily 300 mg kg¹ <i>M. oleifera</i> aqueous extract incorporated with 10 mg kg¹ of F intoxication from the 1st day of gestation until the 20th day. Following the termination of the trial, sera were collected and cerebellar tissue was removed for further examinations, along with the assessment of maternity. <b>Results:</b> The <i>M. oleifera</i> significantly normalized serum FSH, LH, progesterone, dopamine and serotonin levels of F-intoxicated mothers. Additionally, <i>M. oleifera</i> markedly prevented the lipid peroxidation and DNA fragmentation indicated by the tail length and moment in comet assay (-34.4 and -75.3%, respectively, when compared to the fluoride intoxicated group), while sustaining the levels of SOD and CAT revealing its antioxidant activity. The <i>M. oleifera</i> regressed the cerebellar α-amylase (-25.4%) and acetylcholinesterase activity (-40.6%), also attenuated GFAP (-73.4%, p<0.0001), synaptophysin level (216.6%, p<0.0001) and IL-6 expression (-91.2%) comparing to fluoride only treated mothers. <b>Conclusion:</b> Histological and ultrastructural examinations confirmed the recuperating effects of <i>M. oleifera</i> on mothers' cerebellar tissue intoxicated with fluoride indicated by intact folia and restored Purkinje cells number and architecture. The maternal study emphasized the anti-abortifacient activity of moringa against fluoride induced-fetotoxicity.

Synergistic effect between plant extracts and fluoride to protect against enamel erosion: An in vitro study.

Polyphenol-rich solutions, such as plant extracts and teas, can modify the salivary pellicle and improve the protection against dental erosion. In this study, we further explored how these polyphenol-rich plant extracts solutions behave in the presence of fluoride. We distributed enamel specimens into 9 groups (n = 15): Control_No_F- (Deionized water); Control_F- (500 ppm F-), Grape_Seed_No_F- (Grape seed extract), Grape_Seed_F- (Grape seed extract + 500 ppm F-), Grapefruit_Seed_No_F- (Grapefruit seed extract), Grapefruit_Seed_F- (Grapefruit seed extract + 500 ppm F-), Blueberry_No_F- (Blueberry extract), Blueberry_F- (Blueberry extract + 500 ppm F-), and Sn2+/F-_Rinse (commercial solution containing 800 ppm Sn2+ and 500 ppm F-). The specimens were submitted to 5 cycles (1 cycle per day), and each cycle consisted of: salivary pellicle formation (human saliva, 30 min, 37°C), modification of the pellicle (2 min, 25°C), pellicle formation (60 min, 37°C), and an erosive challenge (1 min, citric acid). Between cycles, the specimens were kept in a humid chamber. Relative surface hardness (rSH), relative surface reflection intensity (rSRI) and calcium released to the acid were analysed, using general linear models, and Kruskal-Wallis with post-hoc Dunn's tests. We observed that the presence of fluoride in synergy with the extract solutions provided better protection than the groups containing extract or fluoride only. For rSH, we observed a significant main effect of extracts (F(4,117) = 9.20; p<0.001) and fluoride (F(1,117) = 511.55; p<0.001), with a significant interaction (F(3,117) = 6.71; p<0.001). Grape_Seed_F- showed the best protection, better than fluoride, and Sn2+/F-_Rinse. Calcium results also showed greater protection for the groups containing fluoride, whereas for rSRI, despite a significant interaction between extract and fluoride (F(3,117) = 226.05; p<0.001), the differences between the groups were not as clearly observed. We conclude that polyphenols from plant extracts, when combined with fluoride, improve the protective effect of salivary pellicles against enamel erosion.

Redox and biometal status in Wistar rats after subacute exposure to fluoride and selenium counter-effects.

This study aimed to investigate the effect of 150 mg/L sodium fluoride (NaF) on redox status parameters and essential metals [copper (Cu), iron (Fe), and zinc (Zn)] in the blood, liver, kidney, brain, and spleen of Wistar rats and to determine the protective potential of selenium (Se) against fluoride (F-) toxicity. Male Wistar rats were randomly distributed in groups of five (n=5) receiving tap water (control) or water with NaF 150 mg/L, NaF 150 mg/L + Se 1.5 mg/L, and Se 1.5 mg/L solutions ad libitum for 28 days. Fluorides caused an imbalance in the redox and biometal (Cu, Fe, and Zn) status, leading to high superoxide anion (O2 .-) and malondialdehyde (MDA) levels in the blood and brain and a drop in superoxide dismutase (SOD1) activity in the liver and its increase in the brain and kidneys. Se given with NaF improved MDA, SOD1, and O2 .- in the blood, brain, and kidneys, while alone it decreased SH group levels in the liver and kidney. Biometals both reduced and increased F- toxicity. Further research is needed before Se should be considered as a promising strategy for mitigating F- toxicity.

Dentin Discoloration and Pulpal Ion Concentrations Following Silver Diamine Fluoride and Potassium Iodide Treatment.

OBJECTIVE: The objective of this in vitro study was to evaluate the effects of potassium iodide on dentin discoloration and ion penetration into the pulp chamber after application of silver diamine fluoride (SDF). METHODS: Proximal surfaces of extracted one-rooted sound human teeth were polished to obtain flat dentin and treated with 17% EDTA for two minutes. Each tooth was then fixed to a test tube. The specimens were distributed into five groups according to the treatment: SDF (Advantage Arrest, Elevate Oral Care); SDF-KI (SSKI, Upsher-Smith); RV-SDF (Riva Star, SDI); and RV-SDFKI. The CIE L*a*b* color value for each proximal dentin was assessed using a colorimeter (CR200, Konica-Minolta) at baseline, after two minutes, 10 minutes, and 24 hours, and ΔE compared to baseline was calculated. Subsequently, the water-filled tube was inverted to collect the ions that had penetrated from the tooth surface into the pulp chamber. Silver, iodide, and fluoride in the pulp were measured using trace element analysis (ISMat) and fluoride ion-selective electrode (Orion, Thermo Scientific) at days 1, 2, 7, and 14. RESULTS: Dentin in both SDF-KI groups showed no visual change in color whereas the groups using only SDF exhibited gradual staining. The time, the treatment and their interaction had a significant effect on ΔE (p<0.001). The fluoride concentrations at day 1 and day 14 for group SDF and KI were significantly lower compared to SDF (p=0.044). There was a difference between control groups and other groups in silver and iodine (p<0.05), whereas there was no significant difference among groups with treatment after 14 days (p>0.05). CONCLUSION: The application of KI after SDF treatment could significantly reduce the dentin staining. The pulpal fluoride concentration in the groups using SDF-KI was lower compared to the ones using SDF only, whereas there was no difference observed in iodide and silver among the groups with treatment.

Camellia sinensis Chloroplast Fluoride Efflux Gene CsABCB9 Is Involved in the Fluoride Tolerance Mechanism.

Soil is a main source of fluoride for plants. The tea plants (Camellia sinensis) accumulate excessive amounts of fluoride in their leaves compared to other plants, but their fluoride tolerance mechanism is poorly understood. A chloroplast fluoride efflux gene (CsABCB9) was newly discovered by using transcriptome analysis, cloned from Camellia sinensis, and its function was demonstrated in the fluoride detoxication mechanism in Escherichia coli/Xenopus laevis oocytes and Arabidopsis thaliana. CsABCB9 is expressed in tea leaves upon F− treatment. The growth of tea, E. coli, and Arabidopsis were inhibited by F− treatment. However, growth of CsABCB9-overexpression in E. coli was shown to increase with lower fluoride content under F− treatment compared to the control. Furthermore, chlorophyll, xanthophyll and soluble sugar contents of CsABCB9-overexpression in Arabidopsis were improved under F− treatment compared to the wild type. CsABCB9 functions in fluoride transport, and the mechanism by which CsABCB9 improves fluoride resistance in tea is mainly chloroplast protection through fluoride efflux.

Upregulation of protein N-glycosylation plays crucial roles in the response of Camellia sinensis leaves to fluoride.

The tea plant (Camellia sinensis) is one of the three major beverage crops in the world with its leaves consumption as tea. However, it can hyperaccumulate fluoride with about 98% fluoride deposition in the leaves. Our previously studies found that cell wall proteins (CWPs) might play a central role in fluoride accumulation/detoxification in C. sinensis. CWP is known to be glycosylated, however the response of CWP N-glycosylation to fluoride remains unknown in C. sinensis. In this study, a comparative N-glycoproteomic analysis was performed through HILIC enrichment coupled with UPLC-MS/MS based on TMT-labeling approach in C. sinensis leaves. Totally, 237 N-glycoproteins containing 326 unique N-glycosites were identified. 73.4%, 18.6%, 6.3% and 1.7% of these proteins possess 1, 2, 3, and ≥4 modification site, respectively. 93.2% of these proteins were predicted to be localized in the secretory pathway and 78.9% of them were targeted to the cell wall and the plasma membrane. 133 differentially accumulated N-glycosites (DNGSs) on 100 N-glycoproteins (DNGPs) were detected and 85.0% of them exhibited upregulated expression after fluoride treatment. 78.0% DNGPs were extracellular DNGPs, which belonged to CWPs, and 53.0% of them were grouped into protein acting on cell wall polysaccharides, proteases and oxido-reductases, whereas the majority of the remaining DNGPs were mainly related to N-glycoprotein biosynthesis, trafficking and quality control. Our study shed new light on the N-glycoproteome study, and revealed that increased N-glycosylation abundance of CWPs might contribute to fluoride accumulation/detoxification in C. sinensis leave.

Vitamin C and E supplementation can ameliorate NaF mediated testicular and spermatozoal DNA damages in adult Wistar rats.

OBJECTIVE: Present study was designed to explore the efficacy of vitamin C and E (VC&VE) against fluoride mediated testicular, epididymal and spermatozoal anomalies. MATERIALS AND METHODS: Thirty two adult Wistar rats were divided into four groups. Group-I was control; Group-II received sodium fluoride (NaF) at 15 mg/kg/day dose; Group-III was provided with VC (200 mg/kg/day) and VE (400 mg/kg/day) plus NaF; Group-IV received only VC&VE. Structural integrity and oxidative stress markers (superoxide dismutase, catalase, malondialdehyde and protein carbonyl) of testis and epididymis were assessed. Spermatozoal parameters (count, motility, viability and hypo-osmotic swelling) were evaluated. Testicular functional maker enzymes (acid phosphatase, alkaline phosphatase and lactate dehydrogenase) were also assessed. Integrity of testicular and spermatozoal DNA was evaluated. Testicular fluoride content was measured. RESULT: Fluoride induced structural changes and alterations of oxidative stress markers were observed in testis and epididymis. Spermatozoal potentials were altered and reduced activities of testicular functional marker enzymes were observed. Fluoride caused testicular and spermatozoal DNA damages. VC&VE supplementation resulted in protection from all fluoride mediated alterations and helped in attenuating testicular fluoride accumulation. CONCLUSION: Antioxidant properties of VC&VE ameliorated fluoride mediated reproductive damages but only supplementation did not exhibit any notable effect compared to control rats.

DOES TITANIUM TETRAFLUORIDE PROMOTE A PROTECTIVE EFFECT ON ERODED TOOTH? A SYSTEMATIC REVIEW AND META-ANALYSIS.

OBJECTIVE: This systematic review and meta-analyses aimed to evaluate the potential protective effect of titanium tetrafluoride (TiF4) compound compared to sodium fluoride (NaF) on eroded enamel or dentin. METHODS: Two independent reviewers searched PubMed, Scopus, Web of Science, LILACS, Cochrane, EMBASE, and Open gray Literature up to June 2020. MeSH terms, supplementary concepts, synonyms, and free terms were applied based on PICOS strategy (P- population: eroded enamel or dentin; I- intervention: eroded teeth treated with solution or varnish of TiF4; C- comparison: eroded teeth treated with a solution or varnish of NaF; O- outcome: mineral loss measurement; S- study design: in vitro studies). Seven distinct parameters evaluated the risk of bias, and all studies were submitted to 4 meta-analyses (first and second meta-analyses related to enamel loss treated with fluoride solution or varnish, respectively; third and fourth meta-analyses related to dentin loss treated with fluoride solution or varnish, respectively). A sensitivity test was performed by the RevMan software, based on standard mean difference calculation (P < .05). RESULTS: A total of 10,260 studies were retrieved, from which 22 studies were included in systematic review through eligibility criteria, of which 1 study was classified as low risk of bias, 19 as medium, and 3 as high. Seventeen studies were included in 4 different meta-analyses, presenting considerable overall heterogeneity (I2 = 95; 92; 95; and 93%, respectively) with statistical significance for TiF4 on the first meta-analysis, and no statistical significance for second, third, and fourth meta-analyses. CONCLUSIONS: Titanium tetrafluoride compound delivered as solution provided significant effective protection for erosion lesions on the enamel surface compared to sodium fluoride. However, when TiF4 or NaF were applied on enamel or dentin surfaces as varnish, both provided the same effective protection, based on in vitro studies with considerable heterogeneity.