Search

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

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.

The aim of the present study was to evaluate the effect of commercial sweeteners on root dentin demineralization using a microcosm biofilm model. Bovine dentin specimens with pre-determined surface hardness were randomized into six groups according to the studied sweeteners: sucralose, stevia, saccharin, aspartame. Sucrose was used as a positive control and an untreated group as a negative control. The specimens were submitted to biofilm development from one saliva donor and the cariogenic challenge occurred on subsequent five days, twice a day. At the end, the percentage of surface hardness loss (%SHL) and biomass was determined and submitted to ANOVA followed by Tukey's test. Sucrose presented the highest rate of demineralization, however, all sweeteners tested lead to a statistically higher root demineralization compared to the negative control (p

Objective The aim of this pilot study was to address the reliability, internal validity and viability of oral health methods used in the South American Youth/Child cARdiovascular and Environmental study. Methods South American Youth/Child cARdiovascular and Environmental study was a multicentre feasibility observational study and conducted in seven South American cities. The training sessions were performed in two steps before data collection: the first verified the inter-rater reproducibility between the examiners of the six centres in relation to the gold standard, and the second one verified the inter-rater reproducibility between the examiners at each centre in relation to the main rater. The diagnostic methods used were International Caries Detection and Assessment System II and Pulpal Involvement, Ulceration, Fistula and Abscess for dental caries and Periodontal Screening and Recording and Index Plaque for periodontal disease. Anthropometric variables were measured and used to calculate the body mass index and were classified according to the cut-off points defined by the International Obesity Task Force. Cohen's kappa coefficient and proportions of agreement were calculated to report inter-rater and intra-rater reliability in the calibration process and pilot study. Results The inter-rater weight kappa ranged from 0.78 to 0.88 and proportion of agreement from 96.07% to 98.10% for the International Caries Detection and Assessment System II and for the Periodontal Screening and Recording, 0.68 to 0.95 and 94.40% to 98.33%, respectively, in the calibration process. At the pilot study, a total of 490 children (40.8% overweight and 12.9% obese) and 364 adolescents (23.4% overweight and 4.3% obese) were examined. The prevalence of dental caries was 66% in children and 78% in adolescents, and gingival bleeding was 49% and 58.20%, respectively. Conclusion The results demonstrated good reliability and internal validity after the examiners were trained, as well as the feasibility of using the methods chosen for this multicentre study.

Background There is a lack of information about the association of high-fluoride dentrifrice and fluoride-containing bonding material to prevent enamel white spot lesions development adjacent to brackets. The aim of this in vitro study was to evaluate the effect of high-fluoride dentifrice and fluoride-containing bonding material on enamel demineralization adjacent to orthodontic brackets. Material and methods Forty-eight enamel specimens with 7x7x2 mm were obtained from bovine incisors. Orthodontic brackets were bonded with fluoride-containing resin composite (OrthoCem®) or fluoride-free low viscosity resin. The specimens were submitted to an 8-day pH cycling that consisted in the daily immersion of specimens in the demineralizing solution for 4 h and in artificial saliva for 20 h in an incubator at 37° C. The treatments consisted in 5 min-immersion between the cycles of fluoride (F) suspensions containing 275 µg F/mL, 1,250 µg F/mL or distilled water (negative control). The 275 and 1,250 µg F/mL concentrations were used to simulate salivary dilution of 1,100 and 5,000 µg F/g dentifrices during toothbrushing. After the experiment, cross-sectional hardness was performed to analyze the lesion area of the specimens. Tukey post hoc test after two-way ANOVA with p at 5% was used as statistical analysis. Results The specimens treated with high-fluoride dentifrice showed significantly less demineralization in comparison with the other treatments (p>0.05). There was a significant difference in the cross-sectional hardness values for the specimens bonded with OrthoCem when compared to the low viscosity resin (p>0,05). Conclusions The use of high-fluoride dentifrice associated with fluoride-containing bonding material promoted a greater reduction of enamel demineralization adjacent to orthodontic brackets. Key words:Demineralization, dentifrice, fluoride, bonding materials, orthodontic brackets.

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-1β (p = 0.04, FDR 0.05), and elevated pro-inflammatory (TNF-α,IL-1β,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.

The present study aimed to evaluate the effect of fluoride (F) dentifrice with different F concentrations on root dentine de-/remineralization. Ten healthy volunteers took part in this randomized, double-blinded, cross-over, and split-mouth in situ experimental study. During 4 phases of 7 days, they wore a palatal appliance containing 4 bovine dentine blocks (2 sound and 2 with caries) of 4 × 4 × 2 mm. Treatments were performed with silica-based dentifrices containing 0, 700, 1,300, and 5,000 µg F/g (F as NaF). To provide a cariogenic challenge, a 20% sucrose solution was dripped 3 and 8 times daily on the carious-like and sound blocks, respectively. After each experimental phase, the percentage of surface hardness loss (%SHL) or recovery (%SHR) was calculated and the fluoride concentration in the biofilm was determined. The statistical analysis was performed using ANOVA and the Tukey post hoc test with p at 5%. The relationship between variables was analyzed by linear regression. The results showed a lower %SHL when 5,000 µg F/g dentifrice was used but without a statistically significant difference from the conventional one (1,300 µg F/g). Regarding remineralization and F in biofilms, the high-fluoride dentifrice was expressively superior in mineral replacement on the surface and in the F concentration in the biofilms, respectively, compared to the other 3 products (p < 0.05). Also, a significant linear fit between mineral loss/gain, F in biofilms, and fluoride concentration in the dentifrices could be observed. In conclusion, a dose-response F effect was observed, and the high-fluoride dentifrice was effective in enhancing root dentine remineralization in this short-term in situ study.

Background Composites sorption and solubility can be precursors of several chemical and physical processes, which lead to deleterious effects on the polymer structure. This study evaluated the effect of mouthwashes with and without alcohol on the sorption and solubility of conventional and low viscosity bulk fill resins. Material and Methods Four types of Bulk Fill resins (Filtek™ Bulk Fill, X-tra Fil, Filtek™ Bulk Fill Flow and X-tra Base) were submitted to the following mouthwashes: Listerine Cool Mint and Periogard (containing alcohol) and Listerine Zero and Periogard (alcohol-free). The specimens were stored in the mouthwashes for seven days. Solubility and sorption tests were performed according to ISO 4049. Data were analyzed using two-way-ANOVA, followed by Tukey Test. The data were grouped, and a paired t-test was performed to evaluate the effect of alcohol on the properties studied. The p was fixed at 5%. Results Resins immersed in alcohol-containing mouthwashes had higher values of sorption and solubility, with the highest sorption rate for X-Tra Base in Listerine Cool Mint treatment (p

Objective Probiotics are usually given as living cells, but their effects may be also achieved by postbiotics. We hypothesized that probiotics products (spent media and lysate) altered the response induced by P. gingivalis in gingival epithelial cells (GECS). Methods Immortalized human OBA-9 GECs (∼2,5 × 105cells/well) were challenged with P. gingivalis ATCC33277, and co-infected with L. rhamnosus Lr-32 for 4 h. L. rhamnosus Lr-32 spent medium or cells lysate was added to GECs co-infected with P. gingivalis. Another set of OBA-9 GECs were first exposed to P. gingivalis ATCC 33277 and then to the living probiotic or probiotic products. Transcription of genes encoding inflammatory mediators (IL-1β, TNF-α, IL-6, and CXCL-8) and receptors (TLR2 and TLR4) were evaluated by RT-qPCR. P. gingivalis growth under L. rhamnosus Lr-32 postbiotics was also evaluated. Results L. rhamnosus Lr-32 spent media decreased cell viability, while living cells and cell lysates did not. L. rhamnosus Lr-32 lysate, but not spent media, upregulated transcription of inflammatory mediators (IL-1β, TNF-α, IL-6, and CXCL-8) in GECs infected with P. gingivalis. Transcription of TRL2 was upregulated in all experimental groups compared to control, whereas TLR4 was upregulated by the probiotic or its postbiotics in P. gingivalis infected cells. Spent media and lysates reduced the growth of P. gingivalis. Conclusion L. rhamnosus Lr-32 cell components rather than live probiotic enhanced the expression of inflammatory mediators in P. gingivalis infected gingival epithelial cells. The increased potential of Lr-32 cell lysates to promote immune response to the periodontopathogen may favor pathogen elimination but may also lead to additional deleterious effects of the exacerbated inflammation.

The aim of this in vitro study was to assess the root dentin demineralization caused by a microcosm biofilm model that has been exposed to sucrose in different ways. Materials and Methods: Saliva of two volunteers was inoculated into an artificial medium for biofilm growth and dentin blocks were immersed into these media. Dentin specimens were randomly exposed to one of the five experimental conditions: C (control group - no saliva inoculum or sucrose), 0S (saliva inoculum without sucrose, negative control), 3S (three daily one-minute immersions in 20 % sucrose), 6S (six daily one-minute immersions in 20 % sucrose), and CS (continuously immersed in 5 % sucrose). After five days, biofilm was collected to determine the concentration of intracellular and extracellular polysaccharides and the dentin surface hardness loss (SHL) was measured. The experiment was carried out in triplicate. Results: The dentin SHL was higher in groups that were exposed to sucrose (3S, 6S and CS) and there was a statistically significant difference between all groups (p0.001) and there was no statistically significant difference between the other groups (0S, 3S and 6S) (p>0.005). Conclusion: The microcosm biofilm model developed has the potential to produce root dentin demineralization at different exposures to sucrose.

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. 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%). 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 0.05) for all the variables. 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. Since high-fluoride dentifrice was able to reduce demineralization adjacent to brackets, it can be an option to caries management in orthodontics patients.

Objective. This in vitro study investigated the effect of Fluoride (F), Chlorhexidine (CHX), and their association on the viability of Streptococcus mutans using a biofilm model. Materials and Methods. Biofilms were anaerobically grown on glass slides that were vertically suspended in 24-well plates for 5 days. After 48 h of initial growth, biofilms were treated for the next 72 h, 2x/day with CHX at 0.12% and 2%, F as NaF at 0.08% and 0.4% and their association. Results. CHX treatment decreased the bacteria counts either alone or in association with both F concentrations, when compared with control group and the F treatments alone (p

El objetivo fue asociar el uso de protesis dentales totales y la prevalencia de Candida spp. en ancianos institucionalizados con 60 o mas anos de edad, en una ciudad del Nordeste de Brasil. Se llevo a cabo un estudio del perfil de salud y cuantificacion de Candida spp. por aislamiento con agar Sabouraud. A partir de 181 ancianos institucionalizados, solo 17 (66­84 anos) cumplieron los criterios de inclusion. 47,1 % eran totalmente dependientes y 58,8 % necesito ayuda con la higiene. Los farmacos mas utilizados fueron antihipertensivos. Los resultados mostraron una alta prevalencia de Candida spp. (64,5 %) en las protesis dentales totales de los ancianos institucionalizados y esto tal vez sea un reflejo de la deficiente higiene oral.

OBJECTIVE To evaluate root dentine demineralisation, biomass and loosely bound fluoride (CaF2 ) concentration according to different frequencies of sucrose exposure using a high-fluoride dentifrice. BACKGROUND Although high-fluoride dentifrice has been recommended to arrest root dentine lesions, it is not clear whether it can protect dentine from increased frequencies of sucrose exposure. METHODS An in situ, crossover, split-mouth study was conducted in 3 phases with 7 days each, in which 10 volunteers used a palatal device containing 4 bovine root dentine slabs (2 on each side) with predetermined initial hardness. Cariogenic challenge consisted in dripping a 20% sucrose solution 0 (control), 2, 4, 6, 8 or 10 times/d in each block. Volunteers used high-fluoride dentifrice (NaF, 5000 µg F/g) 3 times/d. After each phase, final hardness was measured and the percentage of surface hardness loss (%SHL) calculated. Also, biomass and CaF2 concentration on dentine were determined. The data were processed and analysed by ANOVA and Tukey test with significance level set at 5%. The relationship between the variables was analysed by linear regression and Pearson correlation (r). RESULTS %SHL and biomass were significantly greater than control for sucrose frequencies higher than 6 times/d (P

This study aimed to evaluate the gastrointestinal absorption and renal excretion of fluoride after the ingestion of high-fluoride dentifrice. Twelve volunteers participated in this in vivo, crossover, and blinded study. In three experimental phases, the volunteers were randomly assigned to one of three treatment groups, who ingested either the following: distilled and deionized water (control), conventional dentifrice (1100 μg/g), or high-fluoride dentifrice (5000 μg/g). Both dentifrices contained fluoride in the form of NaF/SiO2. To determine the rate of fluoride absorption, non-stimulated saliva was collected for up to 120 min after ingestion and the area under the curve of the salivary fluoride concentration was plotted as a function of time and the maximum concentration determined. All urine produced during the 24 h before and after ingestion was collected, and urinary excretion was calculated from the difference between the urinary fluoride concentrations in the two periods. A specific ion electrode coupled to an ion analyzer was used to measure fluoride concentrations. Statistical analysis was performed by ANOVA followed by Tukey’s test with p set at 5%. All measured parameters were highest after the ingestion of the dentifrice with 5000 μg/g (p

Data about total fluoride intake in children living in a tropical semi-arid climate city is scarce, thus we conducted this study. Fifty-eight children aged two to five years, living in a Brazilian tropical city with optimally fluoridated water were selected. Dietary samples were collected using the duplicate diet method on two non-consecutive days in the children's home toothpaste was determined by subtracting the amount of fluoride recovered after brushing from the amount placed on the toothbrush. The mean total dose (SD) of fluoride intake was 0.043(0.016) mg F·kg-1·d-1, with the major (60.6%) contribution from water. The factors associated with the ingestion of fluoride from toothpaste were fluoride concentration of the toothpaste (p = 0.03) and the use of kids toothpaste (p = 0.02). The findings suggest that children have a low fluoride intake, measured by at-home meals and use of fluoride toothpaste; drinking water is the main source of fluoride ingestion.

The aim of the study was to evaluate salivary fluoride (F) availability after toothbrushing with a high-F dentifrice. Twelve adult volunteers took part in this crossover and blind study. F concentration in saliva was determined after brushing with a high-F dentifrice (5000 µg F/g) or with a conventional F concentration dentifrice (1100 µg F/g) followed by a 15 mL distilled water rinse. Samples of nonstimulated saliva were collected on the following times: before (baseline), and immediately after spit (time = 0) and after 1, 2, 3, 4, 5, 10, 15, 20, 30, 45, 60, 90, and 120 min. F analysis was performed with a fluoride-sensitive electrode and the area under curve of F salivary concentration × time (µg F/mL × min−1) was calculated. At baseline, no significant difference was found among dentifrices (P> 0.05). After brushing, both dentifrices caused an elevated fluoride level in saliva; however salivary F concentration was significantly higher at all times, when high-F dentifrice was used (P< 0.01). Even after 120 min, salivary F concentration was still higher than the baseline values for both dentifrices (P< 0.001). High-F dentifrice enhanced the bioavailability of salivary F, being an option for caries management in patients with high caries risk.

The main sources of fluoride intake by children are fluoridated water and toothpaste. Little has been studied regarding fluoride intake from these sources in regions with tropical climates and high temperatures throughout the year. This study aimed to determine the amount of fluoride ingested from diet and tooth brushing by children who live in a city with a tropical climate. Sixty-seven children from Teresina, Piauí, Brazil, took part in this study. The city's water supply was optimally fluoridated. The duplicate-diet method was used to determine the fluoride intake from diet. The intake of fluoride from dentifrice was determined by subtracting the amount of fluoride placed on the toothbrush and that recovered after brushing. The concentration of fluoride was measured using an ion-specific electrode and is expressed as milligrams/kilogram of body weight/day. The mean (±SD) total amount was 0.071 ± 0.036 mg F/kg body weight/day, and the relative contributions of diet and toothpaste were 0.025 ± 0.010 and 0.046 ± 0.035, respectively. The factors associated with fluoride intake from toothpaste were: use of children's toothpaste (p = 0.003), use of large amounts of toothpaste (p < 0.001), and a high frequency of tooth brushing (p = 0.003). Sixty-four percent of children had an intake of less than 0.07 mg F/kg body weight/day, which is considered the upper limit for an aesthetically tolerable fluorosis risk. The results suggest that the amount of fluoride ingested by most children who live in a Brazilian city with a tropical climate is considered safe in terms of the risk of dental fluorosis.

Because dentin is more caries-susceptible than enamel, its demineralization may be more influenced by additional fluoride (F). We hypothesized that a combination of professional F, applied as acidulated phosphate F (APF), and use of 1100-ppm-F dentifrice would provide additional protection for dentin compared with 1100-ppm-F alone. Twelve adult volunteers wore palatal appliances containing root dentin slabs, which were subjected, during 4 experimental phases of 7 days each, to biofilm accumulation and sucrose exposure 8x/day. The volunteers were randomly assigned to the following treatments: placebo dentifrice (PD), 1100-ppm-F dentifrice (FD), APF + PD, and APF+FD. APF gel (1.23% F) was applied to the slabs once at the beginning of the experimental phase, and the dentifrices were used 3x/day. APF and FD increased F concentration in biofilm fluid and reduced root dentin demineralization, presenting an additive effect. Analysis of the data suggests that the combination of APF gel application and daily regular use of 1100-ppm-F dentifrice may provide additional protection against root caries compared with the dentifrice alone.

O objetivo deste estudo foi avaliar a sorção e solubilidade de resinas compostas Bulk Fill expostas a enxaguatórios bucais. Para tal utilizou-se dois tipos de resinas Bulk Fill convencionais (Filtek e Voco) e, a partir destas, confeccionou-se corpos de prova em matriz de teflon, que foram posteriormente randomizados em quatro grupos (n8) correspondentes aos enxaguatórios bucais usados: Listerine Cool Mint e Periogard (contendo álcool em suas composições) e Listerine Zero e Periogard (sem álcool), sendo armazenados nestes por sete dias. Os testes de solubilidade e sorção foram realizados de acordo com a norma ISSO 4049. Os dados foram analisados usando-se ANOVA seguido do teste de Tukey para comparação das médias (a0,05). Além disso, os dados foram agrupados e Teste t pareado foi realizado para verificar o efeito do álcool nas propriedades estudadas. Para a resina Filtek, observou-se que o Listerine Cool Mint e Periogard (com álcool na composição) provocou o maior grau de sorção (p

OBJECTIVE: To evaluate the role of socioeconomic variables and self-perceived oral health in the polarization of caries among adolescents in Santa Bárbara D'Oeste, Brazil. MATERIAL AND METHODS: Cross-sectional study. Sampling was randomized and sample size was defined according to WHO criteria. Two hundred and seventy seven adolescents (15 to 18 year-old) were examined by five trained examiners that assessed DMFT index according to WHO criteria. Self-perceived oral health, access to dental services and socio-demographic variables were self-reported. Student's t tests, chi-square tests, and multivariate logistic regression (with significant caries index (SiC) as the outcome), were performed. RESULTS: Mean DMFT was 5.48 (±4.22) and the proportion of "caries free" subjects was 15.5%. Mean DMFT (9.71±2.85) and mean D (1.67±2.18) of SiC positive subjects were significantly higher than mean DMFT (2.88±2.17) and mean D (0.45±0.87) of SiC negative subjects (p

In situ dental biofilm composition under sugar exposure is well known, but sugar effect on the genotypic diversity of S. mutans in dental biofilm has not been explored. This study evaluated S. mutans genotypic diversity in dental biofilm formed in situ under frequent exposure to sucrose and its monosaccharide constituents (glucose and fructose). Saliva of 7 volunteers was collected for isolation of S. mutans and the same volunteers wore intraoral palatal appliances, containing enamel slabs, which were submitted to the following treatments: distilled and deionized water (negative control), 10% glucose + 10% fructose (fermentable carbohydrates) solution or 20% sucrose (fermentable and EPS inductor) solution, 8x/day. After 3, 7 and 14 days, the biofilms were colleted and S. mutans colonies were isolated. Arbitrarily primed polymerase chain reaction (AP-PCR) of S. mutans showed that salivary genotypes were also detected in almost all biofilm samples, independently of the treatment, and seemed to reflect those genotypes present at higher proportion in biofilms. In addition to the salivary genotypes, others were found in biofilms but in lower proportions and were distinct among treatment. The data suggest that the in situ model seems to be useful to evaluate genotypic diversity of S. mutans, but, under the tested conditions, it was not possible to clearly show that specific genotypes were selected in the biofilm due to the stress induced by sucrose metabolism or simple fermentation of its monosaccharides. A composição do biofilme dental in situ exposto a açúcares é bem conhecida, mas o efeito dos açúcares na diversidade genotípica de S. mutans no biofilme dental ainda não foi explorada. Este estudo avaliou a diversidade genotípica de S. mutans no biofilme dental formado in situ sob frequente exposição à sacarose e seus monossacarídeos constituintes (glicose e frutose). Primeiramente, saliva de voluntários foi coletada para isolamento de S. mutans e os mesmos voluntários usaram um dispositivo intraoral palatino, contendo blocos de esmalte, que foram submetidos 8x/dia aos seguintes tratamentos: água destilada e deionizada (controle negativo), solução de glicose 10% + frutose 10% (carboidratos fermentáveis) e solução de sacarose 20% (fermentável e indutor de PEC). Após 3, 7 e 14 dias, os biofilmes foram coletados e colônias de S. mutans foram isoladas. A técnica de reação em cadeia de polimerase usando primers arbitrários (AP-PCR) demonstrou que o genótipo salivar foi detectado em quase todas as amostras de biofilme, independente do tratamento, e parece refletir aqueles genótipos presentes em maiores proporções no biofilme. Além do genótipo salivar, outros foram encontrados nos biofilmes, mas em uma menor proporção e foram distintos entre os tratamentos. Os dados sugerem que o modelo in situ é útil para a avaliação da diversidade genotípica de S. mutans. Porém, nas condições do presente estudo, não foi possível demonstrar que genótipos específicos foram detectados no biofilme devido ao estresse induzido pelo metabolismo da sacarose ou fermentação de seus monossacarídeos.

Streptococcus mutans é especificamente suprimido pelo tratamento intensivo com clorexidina em gel, mas o tempo de recolonização e o efeito em outras bactérias orais não está totalmente claro. Nesse estudo, a recolonização de estreptococos do grupo mutans foi avaliado em nove voluntários adultos saudáveis, os quais eram altamente colonizados por esse microrganismo. Saliva estimulada foi coletada antes (baseline) e 1, 7, 14, 21 e 28 dias após a aplicação de clorexidina em gel a 1% nos dentes dos voluntários por dois dias consecutivos. Em cada dia, o gel foi aplicado utilizando moldeiras descartáveis por 3 x 5 min com intervalos de 5 min entre cada aplicação. A saliva foi inoculada em ágar sangue para determinação dos microrganismos totais (MT); em mitis salivarius ágar para determinação dos estreptococos totais (ET) e em meio mitis salivarius com bacitracina para determinar a contagem de estreptococos do grupo mutans (EGM). O tratamento com clorexidina foi capaz de reduzir as contagens de EGM e a proporção de EGM em relação aos microrganismos totais (%EGM/MT) (p0,05) após 14 dias para EGM e 21 dias para %EGM/MT. As contagens de MT e ET e a proporção de EGM em relação a estreptococos totais não difereriram estatisticamente do baseline (p>0,05) após o tratamento com clorexidina. Os resultados sugerem que o efeito do tratamento com clorexidina em gel na supressão de estreptococos do grupo mutans é limitado a menos de um mês em indivíduos altamente colonizados. Streptococcus mutans is specifically suppressed by intensive treatment with chlorhexidine gel, but the time for recolonization and the effect on other oral bacteria are not totally clear. In this study, recolonization of mutans streptococci was evaluated in nine healthy adult volunteers, who were highly colonized with this microorganism. Stimulated saliva was collected before (baseline) and at 1, 7, 14, 21 and 28 days after application of 1% chlorhexidine gel on volunteers’ teeth for two consecutive days. On each day, the gel was applied using disposable trays for 3 x 5 min with intervals of 5 min between each application. Saliva was plated on blood agar to determine total microorganisms (TM); on mitis salivarius agar to determine total streptococci (TS) and on mitis salivarius agar plus bacitracin to determine mutans streptococci (MS). Chlorhexidine was capable of reducing the counts of MS and the proportion of MS with regard to total microorganisms (%MS/ TM) (p0.05) after 14 days for MS and 21 days for %MS/TM. The counts of TM and TS and the proportion of MS to total streptococci did not differ statistically from baseline (p>0.05) after chlorhexidine treatment. The results suggest that the effect of chlorhexidine gel treatment on suppression of mutans streptococci is limited to less than a month in highly colonized individuals.

The oral cavity harbors several Streptococcus mutans genotypes, which could present distinct virulence properties. However, little is known about the diversity and virulence traits of S. mutans genotypes isolated in vivo under controlled conditions of high cariogenic challenge. This study evaluated the genotypic diversity of S. mutans isolated from dental biofilms formed in vivo under sucrose exposure, as well as their acidogenicity and aciduricity. To form biofilms, subjects rinsed their mouths with distilled water or sucrose solution 8 times/day for 3 days. S. mutans collected from saliva and biofilms were genotyped by arbitrarily-primed PCR. Genotypes identified in the biofilms were evaluated regarding their ability to lower the suspension pH through glycolysis and their acid susceptibility and F-ATPase activity. Most subjects harbored only one genotype in saliva, which was detected in almost all biofilm samples at high proportions. Genotypes isolated only in the presence of sucrose had higher acidogenicity than those isolated only in the presence of water. Genotypes from biofilms formed with sucrose were more aciduric after 30 and 60 min of incubation at pH 2.8 and 5.0, respectively. The present results suggest that biofilms formed under high cariogenic conditions may harbor more aciduric and acidogenic S. mutans genotypes. A cavidade oral apresenta vários genótipos de Streptococcus mutans, que podem possuir diferentes capacidades de virulência. Entretanto, pouco se sabe sobre a diversidade e virulência de genótipos de S. mutans isolados in vivo sob uma condição controlada de alto desafio cariogênico. Este estudo avaliou a diversidade genotípica de S. mutans identificados no biofilme dental formado in vivo na presença de sacarose, assim como a acidogenicidade e aciduricidade desses genótipos. Para possibilitar formação de biofilme, voluntários bochecharam com água destilada ou solução de sacarose 8x/dia durante 3 dias. S. mutans isolados da saliva e do biofilme dental foram genotipados por PCR com primers-arbitrários. Genótipos isolados do biofilme foram avaliados em relação à habilidade de reduzir o pH da suspensão devido à glicólise, em relação à susceptibilidade a ácidos e também atividade F-ATPase. A maioria dos voluntários apresentou apenas 1 genótipo na saliva, que foram detectados em quase todas as amostras de biofilme em altas proporções. Genótipos isolados somente na presença de sacarose apresentaram maior acidogenicidade do que aqueles genótipos isolados apenas na presença de água. Genótipos de biofilmes formados na presença de sacarose foram mais acidúricos após 30 e 60 min de incubação em pH 2,8 e 5,0, respectivamente. Os resultados do presente estudo sugerem que biofilmes formados sob condição de alto desafio cariogênico podem apresentar genótipos de S. mutans mais acidúricos e mais acidogênicos.

The aim of this study was to investigate the temporal relationship between changes in biofilm composition and enamel demineralization following exposure to sucrose. A crossover blind study was conducted in situ in three phases, during which 12 volunteers, divided into three groups, subjected enamel slabs 8 times/day to water (negative control), 10% glucose + 10% fructose (active control) or 20% sucrose solution. Biofilms accumulated for 3, 7 and 14 days were collected and analyzed biochemically and microbiologically, and mineral loss from enamel (ΔZ) was evaluated. Significantly higher ΔZ was found in the sucrose group after 7 days. However, on the 3rd day, lactobacilli, insoluble extracellular polysaccharide (EPS) and intracellular polysaccharide were significantly higher, and the calcium, inorganic phosphorus and fluoride concentrations in the biofilm were significantly lower in the sucrose group than in the negative controls. The only significant difference compared to glucose + fructose treatment was a higher insoluble EPS concentration. The data suggest that, although sucrose induces significant enamel demineralization only after 7 days of biofilm accumulation, changes in the biofilm composition are observed earlier.

It has been suggested that enamel would resist higher frequencies of sucrose exposure if fluoride from water or dentifrice is being used. However, the effect of increasing frequencies of sugar on dental biofilm composition is not well known. Ten volunteers living in a fluoridated area wore palatal appliances bearing human enamel slabs during 14 days. The slabs were exposed to 20% sucrose solution 0 (control), 2, 4, 6, 8 or 10 times/day and the volunteers used fluoride dentifrice 3 times/day. Enamel demineralization was significantly greater than control for sucrose frequencies higher than 6 times/day. However, biofilm mass, total microbiota, total streptococci, lactobacilli counts and insoluble extracellular polysaccharide concentration increased, while Ca, Pi and F concentration in whole biofilm decreased significantly, with frequencies of sucrose exposure lower than 6 times/day. The findings confirm that fluoride can reduce enamel demineralization if sucrose consumption is not higher than 6 times/day, but changes in the biochemical and microbiological composition of the biofilm are observed with lower frequencies of sucrose use.