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Postbiotic is the term used to define the soluble factors, metabolic products, or byproducts released by live probiotic bacteria or after its lysis. The objective of this study was to carry out the chemical characterization of the postbiotic of Lacticaseibacillus rhamnosus LR-32 and to evaluate its in vitro effect on the development of the Streptococcus mutans biofilm. After the cultivation of the probiotic strain, the postbiotic was extracted by centrifuging the culture and filtering the supernatant. This postbiotic was characterized by using gas chromatography coupled with mass spectrometry (GC-MS), and then it was used to determine the growth inhibition of S. mutans in its planktonic form; additionally, its effects on the following parameters in 48 h biofilm were evaluated: viable bacteria, dry weight, and gene expression of glucosyltransferases and VicR gene. The control group consisted of the biofilm without any treatment. A paired t-test was performed for statistical analysis, with the p-value set at 5%. Seventeen compounds of various chemical classes were identified in the postbiotic, including sugars, amino acids, vitamins, and acids. The treatment with the postbiotic led to an inhibition of the growth of S. mutans in its planktonic form, as well as a decrease in the number of viable bacteria, reduction in dry weight, and a negative regulation of the gene expression of gtfB, gtfC, gtfD, and vicR in its biofilm state, compared with the nontreated group (p < 0.05). The postbiotic of L. rhamnosus impaired the development of S. mutans biofilm.

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Probiotics are beneficial bacteria that may modulate the immune response by altering the maturation and function of antigen-presenting cells, such as dendritic cells. This study aimed to evaluate the antibacterial gene expression of dendritic cells challenged with LPS and probiotics. Immature dendritic cells were obtained from human CD14+ monocytes and challenged with E. coli LPS and probiotics Lacticaseibacillus rhamnosus (LR-32) and Lactobacillus acidophilus (LA-5) at a ratio DC:bacteria of 1:10. The analysis of gene expression was performed by RT-qPCR using the Kit RT2 human antibacterial response. In the supernatant, the cytokines secretion was determined by ELISA. Tukey post-ANOVA with p at 5% was used for statistical analysis. LPS showed the higher upregulation of 29 genes compared with the groups where probiotics were added to LPS, including genes related to an inflammatory response like BIRC3, CASP1, CCL5, CXCL1, IL12B, IL18, MYD88, NLRP3, RIPK1, and TIRAP. Similarly, LPS increased the transcription of genes enrolled with apoptosis such as CARD6, CASP1, IRF5, MAP2K1, MAP2K4, MAPK1, MYD88, NLRP3, RIPK2, TNF, TNFRSF1A, and XIAP when compared to probiotics groups (p < 0.05). Although probiotics decrease several genes upregulated by LPS, the transcription of encoded cytokines IL12A, IL12B, IL1B, IL6, CXCL8, and TNF genes was maintained upregulated by probiotics, except for IL18, which was downregulated by LA-5. LA-5 led to a higher transcription of IL1B, IL6, and CXCL-8 which was followed by the secretion of these proteins by ELISA. The results suggest that probiotics attenuate the transcription of inflammatory and immune response genes caused by LPS.

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OBJECTIVE: The objective of the study was to evaluate the kinetics of salivary F bioavailability after the use of high-fluoride dentifrices with different compositions and their amount of total soluble fluoride (TSF). METHODS: A short-term clinical randomized trial was performed in which 15 adult participants were randomly allocated into three groups: 5000 ppm F-dentifrice, 5000 ppm F-dentifrice + TCP (tri-calcium phosphate) and 1450 ppm F-dentifrice. Unstimulated saliva was collected at different times: baseline (before toothbrushing), immediately after brushing/water rinsing and at 5, 15 and 30 min and 1, 2, 4, 8 and 12 h after brushing. The TSF in dentifrices and saliva samples was analysed using an ion-specific electrode. For statistical analysis, the paired t-test and Kruskal-Wallis were used with Dunn's post-test with a 95% confidence interval. RESULTS: There was no significant difference between the declared TSF and that found in 5000 ppm F-dentifrice and 1450 ppm F-dentifrice (p ≥ 0.13); however, in the 5000 ppm F-dentifrice + TCP, approximately 500 ppm less TSF was observed (p = 0.0024). The area under the curve (AUC, µg F/ml min-1 ) of both high-fluoride dentifrices (321.7 ± 84.0 and 223.6 ± 55.1 for the one without and with TCP, respectively) was higher than the conventional one (89.97 ± 15.6) attesting a higher F-bioavailability (p = 0.04). Furthermore, they were able to provide F-salivary levels higher than the baseline for up to 2 h, while this time was 1 h for the 1450 ppm F-dentifrice (p ≤ 0.003). CONCLUSION: Both high-fluoride dentifrices similarly increased the salivary-F bioavailability in comparison with 1450 ppm F-dentifrice, despite the lower TSF presented by the dentifrice containing TCP.

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Periodontitis and related systemic inflammatory diseases are characterized by imbalanced ratio between pro- and anti-inflammatory factors. Probiotics may control inflammation by altering the inflammatory phenotype of defense cells. We aimed to evaluate the gene transcription of the antibacterial response of monocytes to exposure to probiotic lactobacilli. CD14 + monocytes were obtained by positive selection from peripheral blood mononuclear cells from healthy donors (5 × 104 CD14 + /mL) and cultured with probiotic strains of Lacticaseibacillus rhamnosus (LR-32) and Lactobacillus acidophilus (LA-5) at a 1:10 multiplicity of infection in 24-well plates for 12 h. The gene expression analysis was performed by RT-qPCR using the Kit RT2 human antibacterial response, and in the supernatant, the cytokines were determined by ELISA. Tukey's post hoc test following an ANOVA with a p value of 5% was used for statistical analysis. Both probiotic strains increased the levels of cytokines TNF-α and CXCL-8 in the supernatant compared to the control of non-challenged cells (p < 0.05), but for IL-1Β and IL-6, this effect was observed only for LA-5 (p < 0.05). The fold-regulation values for the following genes for LA-5 and LR-32 were, respectively, IL-12B (431.94 and 33.30), IL-1Β (76.73 and 17.14), TNF-α (94.63 and 2.49), CXCL-8 (89.59 and 4.18), and TLR-2 (49.68 and 3.40). Likewise, most of the other genes evaluated showed greater expression for LA-5 compared to LR-32 (p < 0.05). The positive regulation of inflammatory factors such as IL-1ß promoted by L. acidophilus LA-5 may increase the antibacterial activity of innate defense in periodontal tissues. However, this property may be deleterious by increasing inflammatory response.

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The present study evaluated the effect of high-fluoride dentifrice on dentine demineralization and bacterial composition in a multispecies biofilm model in vitro. A seven-organism bacterial consortium was grown on bovine dentine discs in a high-throughput active attachment model. The biofilms were submitted twice per day to the following dentifrices treatments: 5,000 ppm F, 1,100 ppm F, with placebo as a negative control. After 5 days of biofilm growth, dentine samples were assessed by transversal microradiography, the biofilm was collected for bacterial counts and the pH of the media was determined. Lower integrated mineral loss values were observed when 5,000 ppm F-treatment was used compared to the other treatments. Overall microbiological counts decreased with increasing F-concentration as well the pH of the media throughout the experiment. The 5,000 ppm F-treatment caused a shift in microbial composition and reduced dentine demineralization in the in-vitro experimental model.

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The use of fluoridated dentifrices is recognized as the main reason for the decline of dental caries and its effect is associated with the bioavailability of fluoride (F) in the oral cavity. High-fluoride dentifrice has been indicated for patients at high risk of caries and management of root lesions. This study aimed to evaluate the bioavailability of F in saliva after the use of high-fluoride dentifrice during the nocturnal period. Fifteen healthy adults participated in this is in vivo and crossover study in which the concentration of F in their saliva was determined after brushing with the tested dentifrices: a conventional (1450 ppm F) or with high-fluoride concentration (5000 ppm F). Before brushing, the participants collected the non-stimulated saliva (baseline), immediately after brushing (time zero) and after 5min, 2h, 4h, and 8h, during the nocturnal period (between 10:00 pm and 06:00 am). The salivary F concentration was determined using a specific F ion electrode. Regarding statistical analysis, a paired t-test was used to compare dentifrices with p fixed at 5%. At baseline, there was no significant difference between groups (p>0.001). Immediately after brushing, both dentifrices increased the F salivary concentration, with the highest concentration reached in time zero; however, the use of 5000 ppm F dentifrice maintained the higher F salivary concentration at all times evaluated (p<0.001), remaining higher until 8 h after brushing. Furthermore, this treatment showed higher F bioavailability in relation to time, evaluated by the area under the curve (p<0.001). Thus, it can be concluded that the high-fluoride dentifrice increased the bioavailability of salivary F during the nocturnal period in comparison with conventional dentifrice.

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BACKGROUND: High-fluoride dentifrice is used to manage root caries, but there is no evidence whether its association with nanohydroxyapatite could provide an additional protection for root caries. Therefore, this study aimed to develop and evaluate the effect of an experimental dentifrice with high fluoride (F-) concentration and nanohydroxyapatite (nano-HA) on root dentin demineralization. MATERIALS AND METHODS: After formulation of dentifrices, root dentin specimens were randomly assigned to six groups (n = 10) using different dentifrice treatments: placebo; nano-HA without F-; 1,100 µg F-/g; 1,100 µg F-/g + nano-HA; 5,000 µg F-/g; and 5,000 µg F-/g + nano-HA. A pH cycling model was performed for 10 days, in which treatments were performed twice a day. After that period, the longitudinal hardness was evaluated and the area of demineralization (ΔS) was calculated. The formulated dentifrices were evaluated for primary stability, cytotoxicity, and other technical parameters. Two-way ANOVA and Tukey's test with p set at 5% were used for data analysis. RESULTS: The experimental dentifrices were stable and had no cytotoxicity. Regarding dentin demineralization, the placebo group significantly increased ΔS compared to all other treatment groups (p<0.001). The dentifrices containing 5,000 µg F-/g, regardless of the presence of nano-HA, led to a smaller lesion area in relation to the other treatments (p<0.001). CONCLUSION: The findings of this study suggest that nano-HA reduced dentin demineralization, and dentifrice with 5,000 µg F-/g dentifrices, regardless of the presence of nano-HA, showed a greater reduction in root dentin demineralization.

Assuntos

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Clinical features suggest differences in immune response among periodontitis forms, albeit a large number of cytokines and chemokines remain to be evaluated. The saliva is an available source of mediators and its analysis would be valuable in order to understand pathophysiological differences. The objective of this study was analyze chemokines/cytokines profile in whole saliva of individuals with severe periodontitis (Stage III) presenting moderate [Grade B; GB] or rapid progression rate with a localized incisor-molar pattern [Grade C; GC/IMP]. A case-control study was designed for each periodontitis group. GB (n = 9) and GC/IMP (n = 7) patients and their healthy controls (C-GB, n = 9 and C-GC, n = 7) were evaluated. Non-stimulated saliva samples were assessed by a multiplex assay for a total of 40 cytokines, C-C and C-X-C motif chemokines. GC/IMP group presented higher levels of CCL17 and CCL27 (p = 0.04, FDR > 0.05), and lower levels of CCL2 (p = 0.04, FDR > 0.05) and CCL25 (p = 0.006, FDR < 0.05) when compared to its control. GB patients had higher levels of IL-6, IL-1ß (p = 0.04, FDR > 0.05), and elevated pro-inflammatory (TNF-α,IL-1ß,INF-γ,IL-6, IL-16): anti-inflammatory (IL-2, IL-4, IL-10) ratio (p = 0.01, FDR < 0.05) compared to its control [p-values by Mann-Whitney test, and False Discovery Rate (FDR) by Benjamini-Hochburg corrections]. CCL-chemokines and cytokines contributed to differences between GC/C-GC and GB/C-GB, respectively (p < 0.05, PERMANOVA test). These preliminary data revealed that each periodontitis phenotype presented distinct immune profiles differentially expressed in saliva compared to their related controls, suggesting differences in the etiopathogenesis of GB and GC/IMP.

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OBJECTIVE: This in situ study evaluated the effect of high-fluoride dentifrice (5000 µg F-/g) and fluoride-containing bonding composite resin on enamel demineralization adjacent to orthodontic brackets. METHODS: Ten volunteers wore palatal appliances containing bovine enamel blocks with metallic brackets bonded with fluoride-free or fluoride-containing composite resin. During three phases of 14 days each, three dentifrices with different fluoride concentrations (0, 1100, and 5000 µg F-/g) were tested. The cariogenic challenge consisted of 20% sucrose solution dripped 8x/day onto the dental blocks. At the end of each phase, biofilm formed was collected for fluoride analysis. Cross section hardness was performed in enamel blocks, and the lesion area was calculated. Data were analyzed by two-way ANOVA followed by Tukey post hoc test (α = 5%). RESULTS: The only signicant factor for all the variables under study was the dentifrice. Smaller lesion area and higher fluoride concentration on biofilm were found in 5000 µg F-/g group, irrespective of bonding composite resin (p < 0.001). Neither bracket-bonding composite resin nor the interaction between the factors was statistically significant (p > 0.05) for all the variables. CONCLUSION: High-fluoride dentifrice is effective in reducing demineralization on enamel adjacent to orthodontic brackets, while the fluoride-containing bonding composite resin does not influence it. CLINICAL SIGNIFICANCE: Since high-fluoride dentifrice was able to reduce demineralization adjacent to brackets, it can be an option to caries management in orthodontics patients.

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OBJECTIVE: This study aimed to evaluate F release from GICs before and after recharging with F-dentifrices and after aging process. METHODS: Fifteen specimens of GICs (conventional, resin modified, and high viscosity) and composite resin were stored individually in a polystyrene tube containing 2 ml of deionized water (DW), with water replacement every 24 hours. After 15 days, the specimens were treated with a dentifrice suspension (1 : 3 by volume) containing 0 µg F/g (n = 5), 1,100 µg F/g (n = 5), or 5,000 µg F/g (n = 5). After 3 min, the specimens were rinsed and replaced in new tubes with 2 ml of DW. This procedure was performed 2x/day for 2 days. The readings were taken on days 1, 5, 10, and 15 before and after the treatments. A second experiment was performed, using the same specimens of the previous study that were submitted to an aging process (specimens were kept in 2 ml of DW, remaining at 37°C for 36 weeks). Readings using specific electrode for F detection were taken on days 1, 5, 10, and 15 after treatment of the samples as described above. Data were analyzed by ANOVA and Tukey's test with α fixed at 5%. RESULTS: It was observed that the highest release of F for all the GICs occurred on the first day after the treatments, especially when using a high-fluoride dentifrice, with decreasing release over time. Also, although aged GICs still recharge with F treatments, the amount of F released was lower than fresh materials. CONCLUSION: GICs present a high F recharge and release capacity, especially in the first 24 hours and after the treatment with a high-fluoride dentifrice, even after material aging.