Constanza E. Fernández

Validation of a Cariogenic Biofilm Model to Evaluate the Effect of Fluoride on Enamel and Root Dentine Demineralization

Due to gingival recession both enamel and root dentine are at risk of developing caries. Both tissues are exposed to a similar environment, however there is not a validated model to evaluate the effect of fluoride on these dental substrates simultaneously. Hence, this study aimed to validate a caries model to evaluate the effect of fluoride to prevent demineralization on enamel and root-dentine. Streptococcus mutans UA159 biofilms were formed on saliva-coated bovine enamel and root dentine slabs (n = 12 per group) mounted in the same well of culture plates. The biofilms were exposed 8×/day to 10% sucrose and treated 2×/day with fluoridated solutions containing 0, 150, 450, or 1,350 ppm F; thus, simulating the use of low to high fluoride concentration toothpastes. The pH values of the culture medium was monitored 2×/day as a biofilm acidogenicity indicator. After 96 h, biofilms were collected for fluoride concentration analysis. The percentage of surface hardness loss (%SHL) was calculated for slabs. The fluoride uptake by the enamel and dentine was also determined. The model showed a dose-response because the biofilm and fluoride uptake increased and %SHL decreased at increasing fluoride concentrations (p < 0.05). Fluoride in the biofilm formed on dentine and fluoride uptake by dentine were higher than those for enamel. With the same fluoride concentration treatment, the percentage of reduction of demineralization was lower for dentine than for enamel. In conclusion, the model was validated in terms of a dose-response effect of fluoride on enamel and root dentine. Furthermore, the findings support the clinical data, suggesting that higher fluoride concentrations are necessary to control caries of root dentine than of enamel.

Fluoride content in toothpastes commercialized for children in Chile and discussion on professional recommendations of use

BACKGROUND In Chile, no information is available regarding the soluble fluoride (F) content in the toothpastes commercialized for children and the countrys guidelines recommend the use of F in toothpastes in an age-dependent concentration. No global consensus has been reached on this subject. AIM To determine the soluble F concentration in dentifrices for children sold in Chile and to discuss Chilean guidelines and professional recommendations of use. DESIGN Three samples of twelve different dentifrices were purchased from drugstores. Toothpastes were analysed in duplicate using an ion-specific electrode. The concentrations of total F (TF) and total soluble F (TSF) were determined (μg F/g). RESULTS Measured TF was consistent with that declared by the manufacturer in eight products. Two dentifrices showed lower TF and two higher F concentrations than declared. A toothpaste, marketed as low-F (450 ppm), showed F concentration threefold higher. Most dentifrices exhibited TSF concentrations similar to the TF content, except one sample that displayed considerably lower TSF than TF. Recommendations on F toothpastes use in children widely vary from country to country. CONCLUSIONS Most dentifrices for children match F content in the labelling, but recommendations are not supported by the best evidence available on the benefit/risk of F toothpastes use.

Effect of the Probiotic Lactobacillus rhamnosus LB21 on the Cariogenicity of Streptococcus mutans UA159 in a Dual-Species Biofilm Model

Despite promising results using probiotics, evidence of the preventive effect on enamel demineralization is insufficient and the cariogenic potential of probiotics is still controversial. Probiotics could affect biofilm formation and interfere with adherence, growth or coaggregation with Streptococcus mutans in biofilms. However, most of the studies have been conducted using planktonic bacteria. Hence, the aim of the study was to assess the effect of probiotic bacteria on the cariogenicity of S. mutans using an in vitro biofilm caries model on enamel. Single-species biofilms (S. mutans UA159, SM or Lactobacillus rhamnosus LB21, LB) or dual-species biofilms simultaneously inoculated (SM + LB) or LB inoculated 8 h after SM (SM → LB) were grown for 96 h. Biofilms were formed on bovine enamel saliva-coated slabs of known surface hardness (SH) and immersed in culture media. Biofilms were exposed 8 times per day to 10% sucrose. Medium pH was monitored twice daily as a biofilm acidogenicity indicator. After 96 h, biofilms were collected to determine biomass and bacteria viability. Slab demineralization was calculated as percentage of SH loss (%SHL). Additionally, the model was tested with different concentrations of the initial inoculum (103, 106, 108 cells/ml) and different adhesion times (2 or 8 h). The dual-species biofilm revealed no LB effects on SM cariogenicity, without changes in acidogenicity or %SHL among groups (p > 0.05, n = 12). Lack of activity of LB on SM cariogenicity persisted even when 105 times higher concentration of the probiotic was tested. Coaggregation was not observed. In conclusion, findings suggest that LB does not reduce cariogenicity of SM in a validated experimental caries model.

Effect of 5,000 ppm Fluoride Dentifrice or 1,100 ppm Fluoride Dentifrice Combined with Acidulated Phosphate Fluoride on Caries Lesion Inhibition and Repair

High fluoride dentifrice (FD; 5,000 ppm F) has been recommended to arrest root dentine lesions and to control enamel caries in high-risk patients. Also, standard FD (1,100 ppm F) in combination with professional fluoride application has been recommended to control dentine caries, but the effect of this combination on enamel has been considered modest. Considering the lack of evaluation comparing the use of 5,000 ppm FD (5,000-FD) versus acidulated phosphate fluoride (APF) application combined with 1,100 ppm FD (1,100-FD) on the inhibition and repair of caries lesions in both enamel and dentine, we conducted this in situ, double-blind, crossover study of 3 phases of 14 days. In each phase, 18 volunteers wore palatal appliances containing enamel and root dentine specimens, either sound or carious, to evaluate the effect of the treatments on the inhibition or repair of caries lesions, respectively. The treatments were non-FD (negative control), 5,000-FD, or 1 APF gel application on dental specimens combined with 1,100-FD used twice per day (APF + 1,100-FD). The reduction of demineralization and enhancement of remineralization were assessed by surface and cross-sectional hardness. Fluoride concentration was determined on dental specimens and on the formed biofilm. For enamel, APF + 1,100-FD and 5,000-FD did not differ regarding the inhibition of demineralization and repair of caries lesions. However, for dentine the difference between these treatments was inconclusive because while APF + 1,100-FD was more effective than 5,000-FD in caries lesion reduction and repair, 5,000-FD was more effective than APF + 1,100-FD in the reduction of surface demineralization. Therefore, the findings show that the combination of APF + 1,100-FD is as effective as 5,000-FD in enamel inhibition of demineralization and enhancement of remineralization.

Titratable acidity of beverages influences salivary pH recovery

A low pH and a high titratable acidity of juices and cola-based beverages are relevant factors that contribute to dental erosion, but the relative importance of these properties to maintain salivary pH at demineralizing levels for long periods of time after drinking is unknown. In this crossover study conducted in vivo, orange juice, a cola-based soft drink, and a 10% sucrose solution (negative control) were tested. These drinks differ in terms of their pH (3.5 ± 0.04, 2.5 ± 0.05, and 5.9 ± 0.1, respectively) and titratable acidity (3.17 ± 0.06, 0.57 ± 0.04 and < 0.005 mmols OH- to reach pH 5.5, respectively). Eight volunteers with a normal salivary flow rate and buffering capacity kept 15 mL of each beverage in their mouth for 10 s, expectorated it, and their saliva was collected after 15, 30, 45, 60, 90, and 120 s. The salivary pH, determined using a mini pH electrode, returned to the baseline value at 30 s after expectoration of the cola-based soft drink, but only at 90 s after expectoration of the orange juice. The salivary pH increased to greater than 5.5 at 15 s after expectoration of the cola drink and at 30 s after expectoration of the orange juice. These findings suggest that the titratable acidity of a beverage influences salivary pH values after drinking acidic beverages more than the beverage pH.

Insoluble NaF in Duraphat® May Prolong Fluoride Reactivity of Varnish Retained on Dental Surfaces

There is no consensus about the clinical recommendation of the time that Duraphat® varnish should be maintained on enamel surfaces without suffering mechanical disturbance by the patient. Considering the importance of calcium fluoride (CaF2)-like reservoirs on the anticaries effect of professional fluoride application, an in vitro study was designed to test the reactivity of Duraphat® varnish with enamel forming these reservoirs as a function of time. Since most fluoride in Duraphat® varnish is insoluble to react and form products on enamel, the relative contribution of the varnish soluble and insoluble fluoride fractions to the reactivity was also evaluated. For this, whole-varnish, containing soluble and insoluble fluoride (total fluoride concentration of 23699 ± 384 µg F/g), or centrifuged varnish, containing only soluble fluoride (fluoride concentration of 258 ± 97 µg F/g), were applied in a standardized manner on enamel slabs (n=8/varnish group/time), which were immersed in continuously renewed artificial saliva for up to 36 h. CaF2-like reservoirs formed on enamel by varnish application were extracted using 1 M KOH and fluoride concentration was measured with ion specific electrode. The results were expressed as µg F/cm(2) of enamel area. Whole varnish formed significantly higher fluoride concentration on enamel than centrifuged varnish, reaching maximum concentration at 24 h (22.0 ± 4.5 µg F/cm(2)). Centrifuged varnish reached maximum concentration at 6 h (3.20 ± 0.81 µg F/cm(2)). In conclusion, a longer varnish retention time than the usually recommended could improve the anticaries effect of Duraphat® varnish, allowing that NaF particles, initially insoluble in the varnish matrix, prolong the reactivity with enamel.

Effect of Fluoride-Containing Toothpastes on Enamel Demineralization and Streptococcus mutans Biofilm Architecture

This study aimed to explore the effect of fluoridated toothpastes on biofilm architecture and enamel demineralization in an in vitro biofilm model. Streptococcus mutans was grown on enamel and treated with slurries of commercial toothpastes, containing SnF2 or NaF. Water and chlorhexidine were used as negative and positive controls, respectively. The developed biofilms were imaged and enamel demineralization was measured. SnF2 and NaF toothpaste treatments significantly reduced enamel demineralization, but SnF2 toothpaste was more effective. Only SnF2 toothpaste and chlorhexidine treatments caused reductions on biofilm mass and thickness. In conclusion, this biofilm model was able to differentiate the effects of the SnF2 and NaF toothpastes on biofilm architecture and enamel demineralization.

Concentración de fluoruro en aguas embotelladas comercializadas en Chile: importancia en caries y fluorosis dental

Background: Bottled water consumption has currently increased and their fluoride (F) concentration may not be ideal in terms of caries benefit or risk of fluorosis. While low concentrations would have little anticaries effect, high F concentration would increase the risk of dental fluorosis. Aim: To measure F concentration in bottled waters sold in Chile. Material and methods: Thirty bottles of water were purchased (15 sold as mineral water, six sold as purified water and nine as flavored water). Samples were analyzed in duplicate with a previously calibrated ion-specific electrode. Mean F concentration of each product was calculated and expressed as ppm F (mg F/L). Results: A mean (±SD) concentration of 0.38±0.42, 0.02±0.006 and 0.11±0.18 ppm F for mineral, purified and flavored waters respectively, was found. Three samples were within the optimal F concentration recommended for drinking water in Chile, which ranges from 0.6 to 1.0 ppm F. Two were above such concentration and the others below. Only two waters displayed F concentration information in the label, which was corroborated by the analysis. Conclusions: Only 10% of the bottled waters commercialized in Chile have potential to prevent caries. The F concentration in most of them does not represent an increased risk of fluorosis.

Frequency of Fluoride Dentifrice Use and Caries Lesions Inhibition and Repair

The clinical relevance of the frequency of fluoride dentifrice (FD) use on enamel caries is based on evidence. However, the relative effect of FD on reduction of demineralization or enhancement of remineralization is unknown and the effect of frequency on root dentine caries has not been explored. The aim of this double-blind, crossover, in situ study, which was conducted in 4 phases of 14 days each, was to evaluate the relationship between the frequency of FD use and enamel and root dentine de- and remineralization. Eighteen volunteers wore palatal appliances containing enamel and root dentine slabs, either sound or carious. Biofilm accumulation on the slab surface was allowed, and 20% sucrose solution was dripped 3 or 8 times per day on the carious and sound slabs, respectively. Volunteers used FD (1,100 μg F/g) in the frequencies 0 (fluoride-placebo dentifrice), 1, 2 and 3 times per day. The demineralization and remineralization that occurred in sound or carious slabs was estimated by the percentage of surface hardness loss (%SHL) or recovery (%SHR). Loosely (CaF2) and firmly (FAp) bound fluoride concentrations were also determined. The relationship between the variables was analyzed by linear regression. The %SHL, CaF2 and FAp concentrations were a function of the frequency of FD use for enamel and dentine, but the %SHR was a function of the frequency of FD use only for enamel (p < 0.05). The results suggest that demineralization in enamel and root dentine is reduced in proportion to the frequency of FD use, but for remineralization the effect of the frequency of FD use was relevant only to enamel.

Recent Advances in Remineralization Therapies for Caries Lesions

Remineralization of caries lesions is naturally achieved by salivary ions, and it can be enhanced by external factors or elements such as fluoride. Numerous studies have demonstrated the remineralizing efficacy of fluoride therapies as well as the limitations with some groups of the population. Consequently, developing new remineralization therapies to close this gap in efficacy has been a priority for the last 2 decades. In this review, we summarize and briefly discuss some of the latest advances in remineralization therapies. Most new therapies try to enhance the effect of fluoride by adding other potentially active ingredients to the formulation, such as calcium, phosphate, stannous, xylitol, and arginine. Other remineralization strategies have focused on creating remineralizing scaffolds within the lesions (e.g., self-assembling peptides). While several of the new remineralization strategies have progressed significantly in recent years, for most of them, the evidence is still insufficient to assess their true clinical potential.