Enamel Mineral Loss Research Articles

Probiotic fermented sheep’s milk containing Lactobacillus casei 01: Effects on enamel mineral loss and Streptococcus counts in a dental biofilm model

Abstract The effects of probiotic fermented sheep’s milk containing Lactobacillus casei 01 on enamel demineralization and microorganism counts in a mixed biofilm model were investigated. Enamel blocks (N = 50) were fixed in polystyrene plates in which a mixed biofilm (Streptococcus mutans 25175, Streptococcus parasanguinis 903, and Streptococcus salivarius 8618) was formed. The mature biofilm was treated for five days with the following: (1) sheep’s milk (SM); (2) fermented sheep’s milk with starter culture (FSMS); (3) fermented sheep’s milk with probiotic culture (FSMP); and (4) fermented sheep’s milk with starter and probiotic cultures (FSMSP). A growth control group was also included (brain–heart–infusion medium with inoculum). The percentage of surface hardness loss (%SHL) and total microorganism and Streptococcus counts (log10 colony-forming units/mL) were calculated. In addition, mean internal mineral density loss determination (ΔZ) by micro–computed tomography and descriptive topographic analysis via scanning electronic microscopy (SEM) were performed. No group prevented %SHL (p = 0.168), with a similarity in values (p > 0.05) confirmed in SEM images. No differences were found between the tested groups and growth control group in terms of total microorganism or Streptococcus counts (p > 0.05). In relation to total microorganism reduction, FSMS and FSMP were similar (p = 0.153), presenting a reduced number of microorganisms. Both were different from FSMSP (p

Evaluation of an antibacterial orthodontic adhesive incorporated with niobium-based bioglass: an in situ study

This in situ study aimed to evaluate the antibacterial and anti-demineralization effects of an experimental orthodontic adhesive containing triazine and niobium phosphate bioglass (TAT) around brackets bonded to enamel surfaces. Sixteen volunteers were selected to use intra-oral devices with six metallic brackets bonded to enamel blocks. The experimental orthodontic adhesives were composed by 75% BisGMA and 25% TEGDMA containing 0% TAT and 20% TAT. Transbond XT adhesive (TXT) was used as a control group. Ten volunteers, mean age of 29 years, were included in the study. The six blocks of each volunteer were detached from the appliance after 7 and 14 days to evaluate mineral loss and bacterial growth including total bacteria, total Streptococci, Streptococci mutans, and Lactobacilli. Statistical analysis was performed using GLM model - univariate analysis of variance for microhardness and 2-way ANOVA for bacterial growth (p<0.05). The 20% TAT adhesive caused no difference between distances from bracket and the sound zone at 10-µm deep after 7 and 14 days. After 14 days, higher mineral loss was shown around brackets at 10- to 30-µm deep for TXT and 0% TAT adhesives compared to 20% TAT. S. mutans growth was inhibited by 20% TAT adhesive at 14 days. Adhesive with 20% TAT showed lower S. mutans and total Streptococci growth than 0% TAT and TXT adhesives. The findings of this study show that the adhesive incorporated by triazine and niobium phosphate bioglass had an anti-demineralization effect while inhibiting S. mutans and total Streptococci growth. The use of this product may inhibit mineral loss of enamel, preventing the formation of white spot lesions.

Cytotoxicity of novel fluoride solutions and their influence on mineral loss from enamel exposed to a Streptococcus mutans biofilm

ObjectiveThis study evaluated the cytotoxicity, antimicrobial activity and in vitro influence of new fluoridated nanocomplexes on dental demineralization. DesignThe nanocomplexes hydroxypropyl-β-cyclodextrin with 1% titanium tetrafluoride (TiF4) and γ-cyclodextrin with TiF4 were compared to a positive control (TiF4), a blank control (without treatment) and negative controls (hydroxypropyl-β-cyclodextrin, γ-cyclodextrin, deionized water), following 12- and 72-hour complexation periods. The cytotoxicity was assessed using the neutral red dye uptake assay at T1–15 min, T2–30 min and T3–24 h. A minimum bactericidal concentration (MBC) against Streptococcus mutans (ATCC 25175) was performed. Enamel blocks were exposed to an S. mutans biofilm, and the percentage of surface microhardness loss was obtained. Biocompatibility and microhardness data were analysed using ANOVA/Tukey tests (p < 0.05). ResultsAt T1, the cell viability results of the nanocomplexes were similar to that of the blank control. At T2 and T3, the 72 h nanocomplexes demonstrated cell viability results similar to that of the blank, while the 12 h solutions showed results different from that of the blank (p < 0.05). All fluoridated nanocompounds inhibited S. mutans (MBC = 0.25%), while the MBC of TiF4 alone was 0.13%. All fluoridated compounds presented a percentage of surface microhardness loss lower than that of deionized water (p < 0.05). ConclusionsThe new fluoridated nanocomplexes did not induce critical cytotoxic effects during the experimental periods, whilst they did show bactericidal potential against S. mutans and inhibited enamel mineral loss.

Effect of Violet-Blue Light on Streptococcus mutans-Induced Enamel Demineralization.

Background: This in vitro study determined the effectiveness of violet-blue light (405 nm) on inhibiting Streptococcus mutans-induced enamel demineralization. Materials and Methods: S. mutans UA159 biofilm was grown on human enamel specimens for 13 h in 5% CO2 at 37 °C with/without 1% sucrose. Wet biofilm was treated twice daily with violet-blue light for five minutes over five days. A six-hour reincubation was included daily between treatments excluding the final day. Biofilms were harvested and colony forming units (CFU) were quantitated. Lesion depth (L) and mineral loss (∆Z) were quantified using transverse microradiography (TMR). Quantitative light-induced fluorescence Biluminator (QLF-D) was used to determine mean fluorescence loss. Data were analyzed using one-way analysis of variance (ANOVA) to compare differences in means. Results: The results demonstrated a significant reduction in CFUs between treated and non-treated groups grown with/without 1% sucrose. ∆Z was significantly reduced for specimens exposed to biofilms grown without sucrose with violet-blue light. There was only a trend on reduction of ∆Z with sucrose and with L on both groups. There were no differences in fluorescence-derived parameters between the groups. Conclusions: Within the limitations of the study, the results indicate that violet-blue light can serve as an adjunct prophylactic treatment for reducing S. mutans biofilm formation and enamel mineral loss.

Effect of sodium fluoride on oral biofilm microbiota and enamel demineralization

ObjectiveFluoride is widely used as an anti-caries agent, e.g. in toothpastes and mouth rinses. However, the nature of the anti-caries action is not entirely clear. Mechanisms suspected to explain the cariostatic effect include inhibitory effects on acid formation by bacteria, inhibition of extracellular polysaccharide (EPS) production, inhibition of enamel demineralization and enhancement of remineralizaton or combination thereof. The aim of this study was to examine with the supragingival Zurich in vitro biofilm model the effect of fluoride in NaF formulation, on the microbiota and on demineralization. MethodsBiofilms consisting of Actinomyces oris, Candida albicans, Fusobacterium nucleatum, Streptococcus oralis, Veillonella dispar and Streptococcus sobrinus, were grown anaerobically on sintered hydroxyapatite or bovine enamel disks, exposed to 200, 400, and 1400 ppm of NaF, or 0.1% chlorhexidine (positive control). The biofilms were harvested after 64 h and CFUs were assessed for total bacteria. Demineralization of enamel disks was measured by quantitative light-induced fluorescence. ResultsNaF did not affect the bacterial numbers. No enamel mineral loss was observed at 1400 and 400 ppm of fluoride, whereas the pH of the surrounding medium was increased to 5.5 and 5.0, respectively, compared to the untreated control (pH 4.5 and mineral loss ΔF of −32%). At 1400 ppm NaF the biofilm’s EPS volume was also significantly reduced. ConclusionsAdministration of NaF completely prevented demineralization without affecting biofilm composition and growth. This protective effect may be attributed to the observed decrease in acid production or EPS volume, or to a shift in the de/remineralization balance.

The effect on dental enamel of the frequency of consumption of fluoridated milk with a cariogenic challenge in situ

ObjectivesTo evaluate the effect on enamel of varying the frequency of consumption of 1.0 mg F in milk once per day, twice per day or once every other day under cariogenic challenge in situ. Materials and methodsIn a controlled, randomised, cross-over, single-blind study, 25 subjects wore an intra-oral lower removable appliance with enamel slabs for 21 days during each study arm. Subjects used F-free toothpaste, the cariogenic challenge comprising of five 2 min dippings per day in a 12% sucrose solution. Subjects dipped the appliances in 50 ml of 5.0 ppm fluoridated milk for five minutes during the test period once per day, twice per day, and once every other day and drank 200 ml of the same milk, once per day, twice per day (100 ml each time), or once every other day (200 ml) immediately on re-inserting their appliance in order to replicate topical and systemic effects. Slabs were analysed with surface microhardness (SMH) for protection against further demineralisation and transverse microradiography (TMR) to assess changes in mineralisation. ResultsUsing SMH, 200 ml of 5.0 ppm F milk once per day was significantly better than 100 ml of 5.0 ppm F twice/day (p < 0.05) and 200 ml once every other day, but not significantly. Using TMR there was a statistically significant difference in mineral loss of enamel between baseline and treatment for all groups, but not between groups. ConclusionsDrinking 200 ml of 5.0 ppm milk once per day every day protected enamel against further demineralisation whereas all three frequencies were effective in promoting remineralisation.

A Novel Kinetic Method to Measure Apparent Solubility Product of Bulk Human Enamel.

Introduction: Tooth enamel mineral loss is influenced by its solubility product value, which is fundamental to the understanding of de- and remineralization resulting from a carious or erosive challenge. Published pKsp values for human enamel and hydroxyapatite range from 110 to 126 suggesting a heterogeneous nature of enamel solubility. However, this range of values may also result from the variety of methods used, e.g., some authors reporting values for suspensions of enamel powder and others for bulk enamel. The aim of this study was to develop a method to measure the solubility of bulk human enamel under controlled in vitro conditions simulating demineralization behavior of enamel within the oral environment using scanning microradiography (SMR). SMR was used to monitor real-time changes in enamel demineralization rates at increasing calcium concentrations in a caries simulating demineralization solution until the concentration at which thermodynamic equilibrium between enamel and solution was achieved.Method: 2 mm thick caries free erupted human enamel slabs with the natural buccal surfaces exposed were placed in SMR cells exposed to circulating caries-simulating 2.0 L 0.1 M pH = 4.0 acetic acid, at 25°C. SMR was used to continuously measure in real-time the decrease in mineral mass during the demineralization at 5 different points from on each slab. Demineralization rates were calculated from a linear regression curve of projected mineral mass against demineralization time. Changes in the demineralization rates were monitored following a series of successive increases in calcium (and phosphate at hydroxyapatite stoichiometric ratios of Ca:P 1.67) were added to the demineralizing solution, until demineralization ceased. The pH was maintained constant throughout.Results: Demineralization halted when the calcium concentration was ~30 mM. At higher calcium concentrations, mineral deposition (remineralization) occurred. By comparison with results from speciation software calculations for the calcium phosphate ternary system, this result suggests that the bulk solubility product of enamel (pKspBEnamel) under the conditions used is 121.Discussion: The apparent pKspBEnamel under these conditions was higher than many previous reported values, and much closer to those previously reported for HAp. However, this is a bulk value, and does not reflect that enamel is a heterogeneous material, nor the influence of ionic inclusions.

Dynamic measurement of the optical properties of bovine enamel demineralization models using four-dimensional optical coherence tomography.

Dental enamel mineral loss is multifactorial and is consequently explored using a variety of in vitro models. Important factors include the presence of acidic pH and its specific ionic composition, which can both influence lesion characteristics. Optical coherence tomography (OCT) has been demonstrated as a promising tool for studying dental enamel demineralization. However, OCT-based characterization and comparison of demineralization model dynamics are challenging without a consistent experimental environment. Therefore, an automated four-dimensional OCT system was integrated with a multispecimen flow cell to measure and compare the optical properties of subsurface enamel demineralization in different models. This configuration was entirely automated, thus mitigating any need to disturb the specimens and ensuring spatial registration of OCT image volumes at multiple time points. Twelve bovine enamel disks were divided equally among three model groups. The model demineralization solutions were citric acid (pH 3.8), acetic acid (pH 4.0), and acetic acid with added calcium and phosphate (pH 4.4). Bovine specimens were exposed to the solution continuously for 48 h. Three-dimensional OCT data were obtained automatically from each specimen at a minimum of 1-h intervals from the same location within each specimen. Lesion dynamics were measured in terms of the depth below the surface to which the lesion extended and the attenuation coefficient. The net loss of surface enamel was also measured for comparison. Similarities between the dynamics of each model were observed, although there were also distinct characteristic differences. Notably, the attenuation coefficients showed a systematic offset and temporal shift with respect to the different models. Furthermore, the lesion depth curves displayed a discontinuous increase several hours after the initial acid challenge. This work demonstrated the capability of OCT to distinguish between different enamel demineralization models by making dynamic quantitative measurements of lesion properties. This has important implications for future applications in clinical dentistry.

Novel multifunctional dental cement to prevent enamel demineralization near orthodontic brackets

ObjectivesWhite spot lesions due to biofilm acid-induced enamel demineralization are prevalent in orthodontic treatments. The aim of this study was to develop a novel bioactive multifunctional cement with protein-repellent, antibacterial and remineralizing capabilities, and investigate the effects on enamel hardness and lesion depth in vitro for the first time. Materials and methods2-Methacryloyloxyethyl phosphorylcholine (MPC), dimethylaminohexadecyl methacrylate (DMAHDM), and nanoparticles of amorphous calcium phosphate (NACP) were incorporated into a resin-modified glass ionomer (RMGI). Extracted human premolars had brackets bonded via four groups: (1) Transbond XT (TB), (2) RMGI (GC Ortho LC), (3) RMGI+MPC+DMAHDM, (4) RMGI+MPC+DMAHDM+NACP. Demineralization was induced via a dental plaque microcosm biofilm model. Samples were tested using polarized light microscopy (PLM) for lesion depth. Enamel hardness was tested for different groups. ResultsIncorporating MPC, DMAHDM and NACP did not affect enamel bond strength. “RMGI+MPC+DMAHDM+NACP” group had the least lesion depth in enamel (p<0.05). Groups with NACP had the highest enamel hardness (p<0.05). Mineral loss (ΔS) in enamel for NACP group was about one third that for RMGI control. “RMGI+MPC+DMAHDM” had greater effect on demineralization-inhibition, compared to RMGI and TB controls. “RMGI+MPC+DMAHDM+NACP” was more effective in protecting enamel prisms from dissolution by biofilm acids, compared to RMGI and TB control groups. ConclusionThe Novel “RMGI+MPC+DMAHDM+NACP” cement substantially reduced enamel demineralization adjacent to orthodontic brackets, yielding much less lesion depth and greater enamel hardness under biofilm acid attacks than commercial controls. The clinical significance is that the novel multi-agent (RMGI+MPC+DMAHDM+NACP) method is promising for a wide range of preventive and restorative applications to combat caries.

The performance of probiotic fermented sheep milk and ice cream sheep milk in inhibiting enamel mineral loss

The study aimed to evaluate the effects of two different sheep milk-based food matrices – fermented sheep milk and ice cream – with added probiotic bacteria (Lactobacillus casei 431) on dental enamel subjected to an in vitro highly cariogenic challenge. Sixty enamel blocks were selected and randomly allocated into five treatment groups (n=12): conventional fermented sheep milk (CFSM), probiotic fermented sheep milk (PFSM), conventional sheep milk ice cream (CSMIC), probiotic sheep milk ice cream (PSMIC) and control using deionized water. The blocks were subjected to highly cariogenic pH cycling and the products were applied (5min), in a blinded way, once a day to simulate a daily use for 8 consecutive days. A microhardness test was performed before and after the treatment to estimate the percentage of microhardness surface loss (% SML). Scanning electronic microscopy (SEM) was performed to confirm the mineral loss. All groups had lost microhardness after the experiment. However, CFSM and PFSM exhibited the most positive findings when compared to the control in both ice creams. Scanning electron microscopy showed less mineral loss in CFSM and PFSM compared with CSMIC, PSMIC and control after the cariogenic challenge. Overall, fermented milk decreased mineral loss from enamel subjected to a highly cariogenic challenge, regardless of the presence of probiotics in their composition, which had a higher efficacy compared to ice cream.

In Vitro Acid-Mediated Initial Dental Enamel Loss Is Associated with Genetic Variants Previously Linked to Caries Experience.

We have previously shown that AQP5 and BTF3 genetic variation and expression in whole saliva are associated with caries experience suggesting that these genes may have a functional role in protecting against caries. To further explore these results, we tested ex vivo if variants in these genes are associated with subclinical dental enamel mineral loss. DNA and enamel samples were obtained from 53 individuals. Enamel samples were analyzed for Knoop hardness of sound enamel, integrated mineral loss after subclinical carious lesion creation, and change in integrated mineral loss after remineralization. DNA samples were genotyped for single nucleotide polymorphisms using TaqMan chemistry. Chi-square and Fisher's exact tests were used to compare individuals above and below the mean sound enamel microhardness of the cohort with alpha of 0.05. The A allele of BTF3 rs6862039 appears to be associated with harder enamel at baseline (p = 0.09), enamel more resistant to demineralization (p = 0.01), and enamel that more efficiently regain mineral and remineralize (p = 0.04). Similarly, the G allele of AQP5 marker rs3759129 and A allele of AQP5 marker rs296763 are associated with enamel more resistant to demineralization (p = 0.03 and 0.05, respectively). AQP5 and BTF3 genetic variations influence the initial subclinical stages of caries lesion formation in the subsurface of enamel.

Cariogenic Potential of Sucrose Associated with Maltodextrin on Dental Enamel

Maltodextrin is a hydrolysate of cornstarch and has been widely used in the food industry associated with sucrose. The addition of starch can increase the cariogenic potential of sucrose; however, there are sparse data regarding the cariogenicity of sucrose associated with maltodextrin. Therefore, the aim of this study was to test in situ if maltodextrin could increase the cariogenic potential of sucrose. This was an in situ, randomized, crossover, split-mouth, and double-blind study. Volunteers wore palatal appliances containing bovine enamel blocks for 2 periods of 14 days. They dripped the following solutions on the enamel blocks 8 times per day: deionized distilled water (DDW), maltodextrin (M), sucrose + maltodextrin (S+M), or sucrose (S). At the end of each experimental period, biofilms were collected and analyzed for microbiological (mutans streptococci, lactobacilli, and total microorganisms counts) and biochemical (calcium, inorganic phosphate, fluoride, and insoluble extracellular polysaccharides concentrations) compositions. The enamel demineralization was assessed by microhardness. Treatments S and S+M resulted in a lower inorganic composition and higher concentration of insoluble extracellular polysaccharides in the biofilms, and higher enamel mineral loss compared to DDW and M. It can be concluded that the cariogenic potential of sucrose is not changed when this carbohydrate is associated with maltodextrin (dextrose equivalent 13-17).

Inhibition of Streptococcus mutans Growth and Biofilm Formation by Probiotics in vitro

To exert anticaries effects, probiotics are described to inhibit growth and biofilm formation of cariogenic bacteria such as Streptococcus mutans (SM). We screened 8 probiotics and assessed how SM growth or biofilm formation inhibition affects cariogenicity of probiotic-SM mixed-species biofilms in vitro. Growth inhibition was assessed by cocultivating probiotics and 2 SM strains (ATCC 20532/25175) on agar. Probiotics were either precultured before SM cultivation (exclusion), or SM precultured prior to probiotic cultivation (displacement). Inhibition of SM culture growth was assessed visually. Inhibition of SM biofilm formation on bovine enamel was assessed using a continuous-flow short-term biofilm model, again in exclusion or displacement mode. The cariogenicity of mixed-species biofilms of SM with the most promising growth and biofilm formation inhibiting probiotic strains was assessed using an artificial mouth model, and enamel mineral loss (ΔZ) was measured microradiographically. We found limited differences in SM growth inhibition in exclusion versus displacement mode, and in inhibition of SM 20532 versus 25175. Results were therefore pooled. Lactobacillus acidophilus LA-5 inhibited significantly more SM culture growth than most other probiotics. L. casei LC-11 inhibited SM biofilm formation similarly to other alternatives but showed the highest retention of probiotics in the biofilms (p < 0.05). Mineral loss from SM monospecies biofilms (ΔZ = 9,772, 25th/75th percentiles: 6,277/13,558 vol% × µm) was significantly lower than from mixed-species SM × LA-5 biofilms (ΔZ = 24,578, 25th/75th percentiles: 19,081/28,768 vol% × µm; p < 0.01) but significantly higher than from SM × LC-11 biofilms (ΔZ = 4,835, 25th/75th percentiles: 263/7,865 vol% × µm; p < 0.05). Probiotics inhibiting SM culture growth do not necessarily reduce the cariogenicity of SM-probiotic biofilms. Nevertheless, SM biofilm formation inhibition may be relevant in the reduction of cariogenicity.

An In Vitro Evaluation of Human Enamel Surfaces Subjected to Erosive Challenge After Bleaching.

This study aimed to evaluate whether tooth enamel bleached with hydrogen peroxide (H2 O2 ) is more susceptible to erosion when compared with unbleached tooth enamel; and whether the presence of calcium (Ca) in the bleaching gel influenced this process. Enamel blocks were prepared from human molars, and submitted to surface microhardness analysis (baseline). Blocks were prepared and randomly divided into four treatment groups (n = 20): G1 and G2-bleached with 7.5% H2 O2 , with and without Ca, respectively; G3 and G4-bleached with 35% H2 O2 , with and without Ca, respectively. After bleaching, these groups were submitted to an erosive challenge with 1% citric acid. G5 and G6 (n = 20, each) were the negative (without bleaching) and positive controls (without bleaching, but with erosion), respectively. The percentage of surface hardness loss (%SHL), the 3D non-contact profilometry and Scanning Electron Microscopy (SEM) analyses were performed. G2 showed the highest %SHL after bleaching. G1 presented the lowest %SHL in comparison with G2, G3, G4, and G6 after erosion (p < 0.05), which was confirmed only by the SEM analysis. It is suggested that low concentration of H2 O2 with calcium can be recommended for at-home bleaching agents, which may avoid the mineral loss of bleached enamel after an erosive challenge. Low concentration of H2 02 with calcium can be recommended for at-home bleaching agents, which may avoid the mineral loss of bleached enamel after an erosive challenge. (J Esthet Restor Dent 29:128-136, 2017).

Impact of combined CO2 laser irradiation and fluoride on enamel and dentin biofilm-induced mineral loss.

The caries-protective effects of CO2 laser irradiation on dental enamel have been demonstrated using chemical demineralization models. We compared the effect of CO2 laser irradiation, sodium fluoride, or both on biofilm-induced mineral loss (∆Z) and Streptococcus mutans adhesion to enamel and dentin in vitro. Ground, polished bovine enamel, and dentin samples were allocated to four groups (n=12/group): no treatment (C); single 22,600-ppm fluoride (F) varnish (5% NaF) application; single CO2 laser treatment (L) with short pulses (5μs/λ=10.6μm); and laser and subsequent fluoride treatment (LF). Samples were sterilized and submitted to an automated mono-species S. mutans biofilm model. Brain heart infusion plus 5% sucrose medium was provided eight times daily, followed by rinses with artificial saliva. After 10days, bacterial numbers in biofilms were enumerated as colony-forming units/ml (CFU/ml) (n=7/group). ∆Z was assessed using transversal microradiography (n=12/group). Univariate ANOVA with post hoc Tukey honestly-significant-difference test was used for statistical analysis. Bacterial numbers were significantly higher on dentin than enamel (p<0.01/ANOVA). On dentin, LF yielded significantly lower CFUs than other groups (p=0.03/Tukey), while no differences between groups were found for enamel. The lowest ∆Z in enamel was observed for L (mean/SD 2036/1353vol%×μm), which was not only significantly lower than C (9642/2452vol%×μm) and F (7713/1489vol%×μm) (p<0.05) but also not significantly different from LF (3135/2628vol%×μm) (p>0.05). In dentin, only LF (163/227) significantly reduced ∆Z (p<0.05). CO2 laser irradiation did not increase adhesion of S. mutans in vitro. Laser treatment alone protected enamel against biofilm-induced demineralization, while a combined laser-fluoride application was required to protect dentin.

Sodium trimetaphosphate enhances the effect of 250 p.p.m. fluoride toothpaste against enamel demineralization in vitro.

This in vitro study investigated the effect of sodium trimetaphosphate (TMP), added to toothpaste containing 250 p.p.m. fluoride, on enamel demineralization. Bovine enamel blocks (n = 96) were subjected to five pH cycles over a 7-d period and treatment with suspensions of toothpastes containing 0, 250, 500, and 1,100 p.p.m. fluoride (as sodium fluoride), as well as with 250 p.p.m. fluoride containing TMP at 0.25, 0.5, 1.0, and 3.0%. Treatment with toothpaste suspensions was performed under agitation twice a day, for 1 min. Surface and cross-sectional hardness, and fluoride firmly bound to enamel, were quantified. Data were subjected to one-way anova, followed by Tukey's test. Low-fluoride toothpastes containing TMP at 0.25-1.0% resulted in enamel mineral loss similar to that seen for the toothpaste containing 1,100 p.p.m. fluoride. Also, the addition of TMP to the toothpaste containing 250 p.p.m. fluoride promoted enamel fluoride concentrations similar to those obtained for the 500 p.p.m. fluoride group. The toothpaste containing 250 p.p.m. fluoride and 0.25% TMP led to the lowest mineral loss among all groups. It was concluded that the addition of as little as 0.25% TMP to a toothpaste containing 250 p.p.m. fluoride can reduce enamel demineralization to levels similar to those seen for a conventional toothpaste containing 1,100 p.p.m. fluoride, in vitro.

Effectiveness of a Toothpaste with Low Fluoride Content Combined with Trimetaphosphate on Dental Biofilm and Enamel Demineralization in situ

Objective: The aim of the present study was to evaluate in situ whether a toothpaste with low fluoride associated with sodium trimetaphosphate (TMP) would provide similar effect to that of a 1,100 ppm F toothpaste. Design: This crossover double-blind study consisted of 4 phases (14 days each), during which 10 volunteers wore oral appliances containing 4 enamel bovine blocks. The cariogenic challenge was performed by the application of a 20% sucrose solution (6×/day). The toothpaste treatments (2×/day) were: placebo, 500 ppm F, 500 ppm F plus 1% TMP, and 1,100 ppm F. At the end, enamel mineral loss and biofilm composition were analyzed. Results: The toothpaste with 500 ppm F plus 1% TMP showed the lowest mineral loss (p < 0.05). Regarding the fluoride and calcium concentrations in the enamel and in the biofilm, there were no significant differences between 500 ppm F plus 1% TMP, and 1,100 ppm F toothpastes (p > 0.569), but they were significantly different when compared to toothpaste with 500 ppm F (p < 0.050). Conclusion: The addition of 1% TMP to a low-fluoride toothpaste reduces enamel demineralization in situ similar to a 1,100 ppm F toothpaste.

Does the presence of sucrose in pediatric antibiotics influence the enamel mineral loss and the Streptococcus mutans counts in dental biofilm?

The role of antibiotics containing sucrose on the formation of dental caries is still controversial. This study aimed to investigate the effect of two antibiotics (amoxicillin and potassium clavulanate suspension), with and without sucrose, on human dental hardness and Streptococcus mutans counts in dental biofilm. Primary tooth fragments (n=72) were coated with nail varnish leaving a window of 2.25 mm diameter. Specimens were fixed in 24-well polystyrene plates, containing BHI medium. S. mutans (clinical strains) represented the inoculum to form biofilm on the fragments for 24 h. Twelve fragments were separated for the initial count of microorganisms (baseline). The other fragments were divided into 4 groups (n=12) of treatment: G1 (Clavulin(r)), G2 (Betamox(r)), G3 (chlorhexidine 0.12%), G4 (sucrose 10%). All specimens had their self-control area (covered area). The cross-sectional microhardness (CSMH) was evaluated for each specimen. All the treated groups had a loss of hardness compared to their self-controls (p<0.05). Both drugs inhibited the S. mutans growth and promoted no CSMH difference among them. Both antibiotics eliminated all formed biofilm and did not cause mineral loss from the enamel, regardless the presence of sucrose in its formulation.

Effects of probiotic fermented milk on biofilms, oral microbiota, and enamel.

The aim of this study was to evaluate in vitro and in vivo the effects of 2 brands of probiotic fermented milk on biofilms, oral microbiota, and enamel. For the in situ experiment, ten volunteers wore palatine devices containing four blocks of bovine dental enamel over 3 phases, during which 20% sucrose solution, Yakult® (Treatment A), and Batavito® (Treatment B) were dropped on the enamel blocks. Salivary microbial counts were obtained and biofilm samples were analyzed after each phase. For the in vivo experiment, the same ten volunteers drunk Yakult® (Treatment C) and Batavito® (Treatment D) in two phases. Saliva samples were collected for microbial analysis after each phase. The in situ study showed that in comparison with Treatment A, Treatment B resulted in fewer total cultivable anaerobes and facultative microorganisms in biofilms, higher final microhardness, lower percentage change in surface hardness, and smaller integrated subsurface enamel hardness. In the in vivo study, Treatment D resulted in a reduction in the counts of all microorganisms. The results suggested that the probiotic fermented milk Batavito®, but not Yakult®, reduced the amount of oral microorganisms and mineral loss in bovine enamel.

Analysis of anticaries potential of pit and fissures sealants containing amorphous calcium phosphate using synchrotron microtomography.

The aim of this study was to analyze the anticaries potential of pit and fissure sealants containing amorphous calcium phosphate (ACP) by synchrotron microtomography. Bovine enamel blocks (4×4 mm; n=50) were selected through surface hardness (Knoop) analysis. Slabs were obtained through cross-sections taken 1 mm from the border of the enamel. Five indentations, spaced 100 μm apart, were made 300 μm from the border. Ten specimens were prepared for each tested material (Ultraseal XT plus TM, Aegis, Embrace, Vitremer and Experimental Sealant). The materials were randomly attached to the sectioned surfaces of the enamel blocks and fixed with sticky wax. The specimens were submitted to pH cycling. After that, the surface hardness (SH1) was determined, and the blocks were submitted to synchrotron microcomputed tomography analysis to calculate the mineral concentration (ΔgHAp cm(-3)) at different areas of the enamel. The comparison between the SH1 and ΔgHAp cm(-3) showed a correlation for all groups (r=0.840; p<0.001). The fluoride groups presented positive values of ΔgHAp cm(-3), indicating a mineral gain that was observed mainly in the outer part of the enamel. The ACP showed mineral loss in the outer enamel compared with fluoride groups, although it inhibited the demineralization in the deeper areas of enamel. The combination of two remineralizing agents (fluoride and ACP) was highly effective in preventing demineralization.