Time-lapse confocal microscopy to study in vitro Streptococcus mutans surface colonization.

The cariogenicity of Streptococcus mutans relates to its ability to form biofilms on dental surfaces. The aim of this work was to develop a flowcell system compatible with time-lapse confocal microscopy to compare the adhesion and accumulation of S. mutans cells on surfaces in unsupplemented media against media containing sucrose or sucralose (a non-metabolized sweetener) over a short period of time. Fluorescent S. mutans 3209/pVMCherry was suspended in unsupplemented media or media supplemented with 1% sucrose or 1% sucralose and passed through a 3D-printed flowcell system. Flowcells were imaged over 60 minutes using a confocal microscope. Image analysis was performed, including a newly developed object-movement-based method to measure biomass adhesion. Streptococcus mutans 3209/pVMCherry grown in 1% sucrose-supplemented media formed small, dense, relatively immobile clumps in the flowcell system measured by biovolume, surface area, and median object centroid movement. Sucralose-supplemented and un-supplemented media yielded large, loose, mobile aggregates. Architectural metrics and per-object movement were significantly different (P < 0.05) when comparing sucrose-supplemented media to either unsupplemented or sucralose-supplemented media. These results demonstrate the utility of a flowcell system compatible with time-lapse confocal microscopy and image analysis when studying initial biofilm formation and adhesion under different nutritional conditions.

Effect of pH and titratable acidity on enamel and dentine erosion.

OBJECTIVES: The relative effect of pH and titratable acidity on tooth erosion remains unclear. We determined the effect of both properties on in vivo salivary pH recovery and on enamel and dentine early erosion in situ. METHODS: Solutions simulating acidic beverages with different pHs (2.5 or 3.5) and titratable acidities (0, 25, or 100 mM citric acid) were tested. In an in vivo study (n = 20 participants), the salivary pH was determined before, during, and up to 2 min after exposure to the tested solutions. In situ, 12 participants exposed enamel and root dentine slabs to the tested solutions simulating a beverage consumption; early erosion was assessed by percentage of surface hardness loss (%SHL). Groups were compared by ANOVA (p < 0.05). RESULTS: Saliva pH was lower after exposure to solutions at pH 2.5, irrespective of titratable acidity; pH recovery took longer for solutions with higher titratable acidities, irrespective of their pHs. In situ, the highest %SHL was observed for the solution with lower pH and higher titratable acidities. The addition of citric acid increased the %SHL by 2.5-3 times in enamel, and at least 5 times in dentine. CONCLUSIONS: Both pH and titratable acidity may play a role on the erosive potential of acidic beverages. CLINICAL RELEVANCE: Acidic beverages with lower pHs promote erosion by an initial acid etching of the surface; those with a higher titratable acidity slow down the salivary pH recovery. Both properties contribute to the overall erosive potential.

On the release of fluoride from biofilm reservoirs during a cariogenic challenge: an in situ study.

Biofilm fluoride reservoirs may be a source of fluoride to the fluid phase during a sugar challenge reducing tooth mineral loss. However, the evidence for that is conflicting and has not been studied in biofilms containing different fluoride levels. In order to test fluoride release from biofilms with distinct fluoride concentrations, biofilms were grown in situ exposed to a combination of placebo, calcium and fluoride rinses forming biofilms with no (fluoride-free rinses), low (fluoride-only rinses) or high (calcium followed by fluoride rinses) fluoride concentrations, and collected before and 5 min after a sucrose challenge. Rinsing with fluoride increased fluoride concentration in the biofilm (p < 0.05), mainly when a calcium pre-rinse was used before the fluoride (p < 0.05). However, after a sugar challenge, no significant increase in the biofilm fluid fluoride concentration was observed, even in the fluoride-rich biofilms (p > 0.05). Fluoride-rich biofilms do not release fluoride to the fluid phase during a sugar challenge.

Fluoride Increase in Saliva and Dental Biofilm due to a Meal Prepared with Fluoridated Water or Salt: A Crossover Clinical Study.

Knowledge about fluoride delivery to oral fluids from foods cooked with fluoridated water and salt is scarce, and no study has evaluated fluoride concentrations in saliva or biofilm during meal consumption. In this randomized double-blind crossover study, 12 volunteers ingested meals (rice, beans, meat, and legumes) prepared with nonfluoridated water and salt (control group), fluoridated water (0.70 mg F/L; water group), and fluoridated salt (183.7 mg F/kg; salt group). Whole saliva was collected before meal ingestion, during mastication, and up to 2 h after meal ingestion. Dental biofilm was collected before and immediately after meal ingestion. Fluoride concentrations in saliva and dental biofilm were determined by an ion-specific electrode. The mean (±standard deviation; n = 4) fluoride concentrations in meals prepared for the control, water, and salt groups were 0.039 ± 0.01, 0.43 ± 0.04, and 1.71 ± 0.32 µg F/g, respectively. The three groups had significantly different fluoride concentrations in saliva collected during mastication (p < 0.0001) and after meal ingestion (p < 0.04; salt > water > control). The fluoride concentration in saliva returned to baseline 30 min after meal ingestion in the water group but remained high for up to 2 h in the salt group (p = 0.002). The fluoride concentration in biofilm fluid differed only between the salt and control groups (p = 0.008). The mastication of foods cooked with fluoridated water and salt increases fluoride concentrations in oral fluids and may contribute to the local effect of these community-based fluoride interventions on caries control.

Mechanical and chemical plaque control in the simultaneous management of gingivitis and caries: a systematic review.

AIM: To report the evidence on the effect of mechanical and/or chemical plaque control in the simultaneous management of gingivitis and caries. MATERIAL AND METHODS: A protocol was designed to identify randomized (RCTs) and controlled (CCTs) clinical trials, cohort studies and prospective case series (PCS), with at least 6 months of follow-up, reporting on plaque, gingivitis and caries. Relevant information was extracted from full papers, including quality and risk of bias. Meta-analyses were performed whenever possible. RESULTS: After the screening of 1,373 titles, 15 RCTs, 10 CCTs and 2 PCS were included. Low to moderate evidence support that combined professional and self-performed mechanical plaque control significantly reduces standardized plaque index [n = 4; weighted mean difference (WMD) = 1.294; 95% CI (0.445; 2.144); p = 0.003] and gingivitis scores [n = 4; WMD = 1.728; 95% CI (0.631; 2.825); p = 0.002]. The addition of fluoride to mechanical plaque control is relevant for caries management [n = 5; WMD = 1.159; 95% CI (0.145; 2.172); p = 0.025] while chlorhexidine rinses are relevant for gingivitis. CONCLUSION: Mechanical plaque control procedures are effective in reducing plaque and gingivitis. The addition of fluoride to mechanical plaque control is significant for caries management. Chlorhexidine rinse has a positive effect on gingivitis and inconclusive role in caries.

Prevention and control of dental caries and periodontal diseases at individual and population level: consensus report of group 3 of joint EFP/ORCA workshop on the boundaries between caries and periodontal diseases.

BACKGROUND: The non-communicable diseases dental caries and periodontal diseases pose an enormous burden on mankind. The dental biofilm is a major biological determinant common to the development of both diseases, and they share common risk factors and social determinants, important for their prevention and control. The remit of this working group was to review the current state of knowledge on epidemiology, socio-behavioural aspects as well as plaque control with regard to dental caries and periodontal diseases. METHODS: Discussions were informed by three systematic reviews on (i) the global burden of dental caries and periodontitis; (ii) socio-behavioural aspects in the prevention and control of dental caries and periodontal diseases at an individual and population level; and (iii) mechanical and chemical plaque control in the simultaneous management of gingivitis and dental caries. This consensus report is based on the outcomes of these systematic reviews and on expert opinion of the participants. RESULTS: Key findings included the following: (i) prevalence and experience of dental caries has decreased in many regions in all age groups over the last three decades; however, not all societal groups have benefitted equally from this decline; (ii) although some studies have indicated a possible decline in periodontitis prevalence, there is insufficient evidence to conclude that prevalence has changed over recent decades; (iii) because of global population growth and increased tooth retention, the number of people affected by dental caries and periodontitis has grown substantially, increasing the total burden of these diseases globally (by 37% for untreated caries and by 67% for severe periodontitis) as estimated between 1990 and 2013, with high global economic impact; (iv) there is robust evidence for an association of low socio-economic status with a higher risk of having dental caries/caries experience and also with higher prevalence of periodontitis; (v) the most important behavioural factor, affecting both dental caries and periodontal diseases, is routinely performed oral hygiene with fluoride; (vi) population-based interventions address behavioural factors to control dental caries and periodontitis through legislation (antismoking, reduced sugar content in foods and drinks), restrictions (taxes on sugar and tobacco) guidelines and campaigns; however, their efficacy remains to be evaluated; (vii) psychological approaches aimed at changing behaviour may improve the effectiveness of oral health education; (viii) different preventive strategies have proven to be effective during the course of life; (ix) management of both dental caries and gingivitis relies heavily on efficient self-performed oral hygiene, that is toothbrushing with a fluoride-containing toothpaste and interdental cleaning; (x) professional tooth cleaning, oral hygiene instruction and motivation, dietary advice and fluoride application are effective in managing dental caries and gingivitis. CONCLUSION: The prevention and control of dental caries and periodontal diseases and the prevention of ultimate tooth loss is a lifelong commitment employing population- and individual-based interventions.

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 x2192; 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.

Oral fluoride levels 1 h after use of a sodium fluoride rinse: effect of sodium lauryl sulfate.

Increasing the concentration of free fluoride in oral fluids is an important goal in the use of topical fluoride agents. Although sodium lauryl sulfate (SLS) is a common dentifrice ingredient, the influence of this ion on plaque fluid and salivary fluid fluoride has not been examined. The purpose of this study was to investigate the effect of SLS on these parameters and to examine the effect of this ion on total (or whole) plaque fluoride, an important source of plaque fluid fluoride after a sufficient interval following fluoride administration, and on total salivary fluoride, a parameter often used as a surrogate measure of salivary fluid fluoride. Ten subjects accumulated plaque for 48 h before rinsing with a 12 mmol/l NaF (228 µg/g F) rinse containing or not containing 0.5% (w/w) SLS. SLS had no statistically significant effect on total plaque and total saliva fluoride but significantly increased salivary fluid and plaque fluid fluoride (by 147 and 205%, respectively). These results suggest that the nonfluoride components of topical agents can be manipulated to improve the fluoride release characteristics from oral fluoride reservoirs and that statistically significant change may be observed in plaque fluid and salivary fluid fluoride concentrations that may not be observed in total plaque and total saliva fluoride concentrations.

Measurement and analysis of microbial fluoride resistance in dental biofilm models.

The interactions between communities of microorganisms inhabiting the dental biofilm is a major determinant of oral health. These biofilms are periodically exposed to high concentrations of fluoride, which is present in almost all oral healthcare products. The microbes resist fluoride through the action of membrane export proteins. This chapter describes the culture, growth and harvest conditions of model three-species dental biofilm comprised of cariogenic pathogens Streptococcus mutans and Candida albicans and the commensal bacterium Streptococcus gordonii. In order to examine the role of fluoride export by S. mutans in model biofilms, procedures for generating a strain of S. mutans with a genetic knockout of the fluoride exporter are described. We present a case study examining the effects of this mutant strain on the biofilm mass, acid production and mineral dissolution under exposure to low levels of fluoride. These general approaches can be applied to study the effects of any gene of interest in physiologically realistic multispecies oral biofilms.

Fluoride export is required for competitive fitness of pathogenic microorganisms in dental biofilm models.

Microorganisms resist fluoride toxicity using fluoride export proteins from one of several different molecular families. Cariogenic species Streptococcus mutans and Candida albicans extrude intracellular fluoride using a CLCF F-/H+ antiporter and FEX fluoride channel, respectively, whereas commensal eubacteria, such as Streptococcus gordonii, export fluoride using a Fluc fluoride channel. In this work, we examine how genetic knockout of fluoride export impacts pathogen fitness in single-species and three-species dental biofilm models. For biofilms generated using S. mutans with genetic knockout of the CLCF transporter, exposure to low fluoride concentrations decreased S. mutans counts, synergistically reduced the populations of C. albicans, increased the relative proportion of commensal S. gordonii, and reduced properties associated with biofilm pathogenicity, including acid production and hydroxyapatite dissolution. Biofilms prepared with C. albicans with genetic knockout of the FEX channel also exhibited reduced fitness in the presence of fluoride, but to a lesser degree. Imaging studies indicate that S. mutans is highly sensitive to fluoride, with the knockout strain undergoing complete lysis when exposed to low fluoride for a moderate amount of time, and biochemical purification the S. mutans CLCF transporter and functional reconstitution establishes that the functional protein is a dimer encoded by a single gene. Together, these findings suggest that fluoride export by oral pathogens can be targeted by specific inhibitors to restore biofilm symbiosis in dental biofilms, and that S. mutans is especially susceptible to fluoride toxicity.

Fluoride export is required for the competitive fitness of pathogenic microorganisms in dental biofilm models.

Microorganisms resist fluoride toxicity using fluoride export proteins from one of several different molecular families. Cariogenic species Streptococcus mutans and Candida albicans extrude intracellular fluoride using a CLCF F-/H+ antiporter and FEX fluoride channel, respectively, whereas oral commensal eubacteria, such as Streptococcus gordonii, export fluoride using a Fluc fluoride channel. In this work, we examine how genetic knockout of fluoride export impacts pathogen fitness in single-species and three-species dental biofilm models. For biofilms generated using S. mutans with the genetic knockout of the CLCF transporter, exposure to low fluoride concentrations decreased S. mutans counts, synergistically reduced the populations of C. albicans, increased the relative proportion of oral commensal S. gordonii, and reduced properties associated with biofilm pathogenicity, including acid production and hydroxyapatite dissolution. Biofilms prepared with C. albicans with genetic knockout of the FEX channel also exhibited reduced fitness in the presence of fluoride but to a lesser degree. Imaging studies indicate that S. mutans is highly sensitive to fluoride, with the knockout strain undergoing complete lysis when exposed to low fluoride for a moderate amount of time. Biochemical purification of the S. mutans CLCF transporter and functional reconstitution establishes that the functional protein is a dimer encoded by a single gene. Together, these findings suggest that fluoride export by oral pathogens can be targeted by specific inhibitors to restore biofilm symbiosis in dental biofilms and that S. mutans is especially susceptible to fluoride toxicity. IMPORTANCE: Dental caries is a globally prevalent condition that occurs when pathogenic species, including Streptococcus mutans and Candida albicans, outcompete beneficial species, such as Streptococcus gordonii, in the dental biofilm. Fluoride is routinely used in oral hygiene to prevent dental caries. Fluoride also has antimicrobial properties, although most microbes possess fluoride exporters to resist its toxicity. This work shows that sensitization of cariogenic species S. mutans and C. albicans to fluoride by genetic knockout of fluoride exporters alters the microbial composition and pathogenic properties of dental biofilms. These results suggest that the development of drugs that inhibit fluoride exporters could potentiate the anticaries effect of fluoride in over-the-counter products like toothpaste and mouth rinses. This is a novel strategy to treat dental caries.

Surface porosity of enamel white spot lesions and penetration of fluorapatite nanocrystals into their subsurface: proof-of-concept study.

OBJECTIVES: In this study, we used atomic force microscopy (AFM) to quantify the size of surface pore apertures of enamel white spot lesions and then demonstrated the penetration of fluorapatite nanocrystals (nFA) into the subsurface of these lesions. METHODS: For the porosity study, enamel lesions were created on three sound human teeth using a demineralizing gel for 8 days. The interface between sound enamel and the artificial lesion was analyzed by AFM. To visualize the penetration of nFA tagged with a calcium-binding fluorophore (Fluo-4) into the subsurface of white spot lesions, we used two-photon microscopy. Sixteen extracted human teeth with either active, natural, or in vitro-created carious lesions in enamel were randomly divided into three groups. The teeth were treated for 2 min with either a suspension of tagged nFA crystals, Fluo-4 alone, or deionized water, and left for 30 min before being washed with distilled water and examined microscopically. RESULTS: A greater concentration of surface pores with larger areas was observed on the in vitro demineralized enamel (29 % of pores greater than 1.0 µm2) when compared with the adjacent sound enamel (8 % of pores greater than 1.0 µm2) (p=0.012, Fisher exact test). In vitro and natural lesions treated with tagged nFA showed fluorescence at depths ranging from 50 to 170 µm, demonstrating penetration of the nFA into the lesion subsurface. The lesions treated with Fluo-4 alone with no crystals showed mostly surface fluorescence (restricted to the outer 25 µm), while those treated with deionized water showed minimal (restricted to the outer 20 µm) to no fluorescence. CONCLUSION: We have demonstrated the use of AFM to quantify the surface pore apertures and two-photon microscopy to visualize nFA crystals in the subsurface of non-cavitated enamel lesions. CLINICAL SIGNIFICANCE: The restoration of the subsurface of non-cavitated caries lesions is a clinical challenge. This study demonstrated that a 2 min application of nFA could penetrate through the surface apertures of non-cavitated enamel lesions into their subsurface.

Chapter 9.1: The Use of Fluorides in the Control of Coronal Caries.

Fluoride is the main agent used to control dental caries, with a very successful history after its widespread implementation worldwide, resulting in significant caries declines around the globe. In this chapter, the mechanism of action and peculiarities of different methods of fluoride use are revised. Fluoride controls caries progression by interfering with the processes of mineral exchanges (demineralization, remineralization) to which teeth are frequently subjected upon exposure to dental biofilm and a diet rich in fermentable sugars. Because caries is a ubiquitous disease of modern societies, different strategies have been developed to successfully keep fluoride available in oral fluids: (1) community-based approaches, such as water fluoridation, in which fluoride is delivered to communities at very low levels; (2) individual-based approaches, such as fluoride toothpastes, in which fluoride is delivered at higher concentrations, directly into the oral cavity, in a convenient combination with the act of brushing teeth to remove dental biofilm; (3) professionally-delivered approaches, such as fluoride gels and varnishes, in which very-high concentration products react with teeth resulting in local fluoride reservoirs that release the ion for weeks/months; and (4) combinations of these approaches, such as the concomitant use of fluoridated water and toothpaste, or the use of professional fluorides in caries-active patients. In conclusion, this chapter reviews the different modalities of fluoride use for coronal caries control, and summarizes the combined application of these methods according to the best evidence available.

Teledentistry adoption and applications: An American Dental Association Clinical Evaluators Panel survey.

BACKGROUND: The rise of teleworking technologies has affected various industries, including dentistry. Although some dentists have used it for some time, many are discovering the benefits of incorporating this technology to complement their existing patient care capabilities. METHODS: To assess how clinicians are using teledentistry in their practices, an electronic survey was developed and deployed to the American Dental Association Clinical Evaluators (ACE) Panel on February 27, 2023. The survey link remained open for 2 weeks. Nonrespondents received reminders after 1 week. RESULTS: Of the 244 respondents (24% response rate), 30% use teledentistry in their practices, with more than one-half of those using synchronous (53%) or asynchronous teledentistry (63%). The most common reasons for incorporating teledentistry were increased convenience for patients (53%), COVID-19 (50%), and increased accessibility to providers (39%). Teledentistry can help serve patients of all ages (the lowest represented age group [0-5 years] had 42% of dentists treating them) and distances, with 63% of teledentistry patients fewer than 20 miles away. Most users adopted teledentistry within the past 3 years and use it fewer than 5 hours per month. Benefits cited include a reduced number of in-person patient visits (63%) and increased access and quality of care (57%). Among nonusers, 60% felt there was no need, and 39% had concerns with reimbursement. CONCLUSIONS: Teledentistry has gained popularity in since the onset of the COVID-19 pandemic but may be underused, despite its potential to benefit a wider range of patients and applications than many believe. PRACTICAL IMPLICATIONS: Education on the capabilities and benefits of teledentistry may help increase adoption and improve patient care.

Early occlusal caries detection using targeted fluorescent starch nanoparticles.

OBJECTIVES: We have previously shown fluorescent cationic starch nanoparticles (FCSNs) penetrate enamel surface porosity of active carious lesions, potentially aiding their detection. Here, we evaluate the in vitro diagnostic accuracy of FCSNs in detecting occlusal caries compared to histologic reference standard. METHODS: 100 extracted human teeth were selected with sound (50), or either non-cavitated (25) or cavitated (25) lesions. A region of interest (ROI) on the occlusal surface was assessed for fluorescence by two independent examiners, after immersion in FCSN solution, water rinse, and illumination by dental curing lamp viewed through orange UV-filter glasses. ROIs were sectioned and evaluated by histology (Downer Criteria) as a gold standard for caries presence. Cohen's Kappa was determined for inter- and intra-examiner agreement, and sensitivity, specificity, and area under the curve of Receiver Operator Curves (ROCAUC) were calculated. The analysis was repeated for the subset of "early" lesions, defined as being limited to enamel. RESULTS: FCSN use resulted in substantial inter-user (k=0.74±0.07), and high intra-user agreement (k=0.80±0.06; 0.94±0.03, by examiner). Sensitivity, specificity and ROCAUC for FCSNs were 88.9%; 94.6%; 0.92±0.06 for all, and 76.9%, 94.6%, and 0.86±0.10 for early lesions. In post hoc analysis, sensitivity seemed to be greater with the FCSN than the expert visual exam, particularly for early lesions. CONCLUSIONS/CLINICAL SIGNIFICANCE: FCSNs are a reproducible and accurate novel technology for occlusal caries detection, with high sensitivity and specificity compared to histology. Future clinical validation is necessary. FCSNs can improve early caries detection and shift treatment towards non-invasive approaches, improving oral health.

Low-fluoride toothpaste and deciduous enamel demineralization under biofilm accumulation and sucrose exposure.

Because low-fluoride toothpaste (500 p.p.m. F) has not clearly been shown to be effective for controlling caries in caries-active children, this experimental in situ study was conducted to evaluate whether its effect, when compared with a conventional toothpaste (1,000-1,500 p.p.m. F), would depend on the cariogenic challenge. During four phases of 14 d each, 14 volunteers used 500 or 1,100 p.p.m. F toothpaste and wore palatal appliances containing deciduous enamel slabs, on which biofilm was accumulated and exposed to 20% sucrose solution at frequencies increasing from two to eight times per day. The F concentration was determined in the biofilm formed, and enamel demineralization was assessed by surface hardness loss (% SHL) and integrated area of hardness x lesion depth (DeltaS). The F uptake by enamel was also determined. Fluoride in biofilm fluid and solids was statistically higher when conventional F toothpaste was used. The toothpastes did not differ statistically in terms of % SHL, DeltaS, and F in enamel, but only the conventional F toothpaste significantly reduced caries-lesion progression according to the frequency of sucrose exposure. The findings suggest that the high-F availability in biofilm, resulting from the use of conventional toothpaste compared with low-F toothpaste, may be important for reducing caries-lesion progression in conjunction with a high frequency of sucrose exposure.

Protocols to Study Dental Caries In Vitro: Microbial Caries Models.

Caries lesions result from the interaction between dental biofilm and sugars. Since the biofilm is an important component in the etiology of the disease, biofilm models have been developed to study the cariogenicity of dietary sugars, as well as the anticaries effect of substances. Two of such models, termed as "static" or "continuous flow," are described in details here together with their advantages, limitations, and applications.

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.

Laboratory and human studies to estimate anticaries efficacy of fluoride toothpastes.

Much more than mechanical biofilm removal, toothbrushing with fluoride toothpastes is an effective way of increasing the availability of fluoride in the oral cavity to reduce demineralization and enhance remineralization of enamel and dentine. These effects of fluoride toothpastes have been estimated by a wide range of laboratory and human studies, which have helped to develop anticaries effective formulations and understand their mechanism of action. These studies have focused on the availability of fluoride in the toothpaste formulations, its bioavailability in saliva and remnants of disturbed biofilm, its reaction with the dental substrate to form loosely bound reservoirs as well as the ultimate reduction of mineral loss and increase in mineral and fluoride content of caries lesions. The specifics of these modes of action and their application in in vitro, in situ and in vivo preclinical tests is presented and discussed.

Summary of the IADR Cariology Research, Craniofacial Biology, and Mineralized Tissue Groups Symposium, Iguaçu Falls, Brazil, June 2012: Gene-environment Interactions and Epigenetics in Oral Diseases: Enamel Formation and its Clinical Impact on Tooth Defects, Caries, and Erosion.

Characteristics of enamel may influence or modulate individual susceptibility to caries and erosion. These characteristics are defined during development, which is under strict genetic control, but can easily be modified in many ways by environmental factors. In the symposium, translational aspects of embryology, biochemistry, and genetics of amelogenesis were presented. The symposium provided unique insight into how basic sciences integrate with clinically relevant problems. The need for improved understanding of risks at the individual level, taking into consideration both environmental exposures and genetic background, was presented. The symposium was divided into four stepwise and interconnected topics as follows: 1) The Many Faces of Enamel Development; 2) Enamel Pathogenesis: Biochemistry Lessons; 3) Environmental Factors on Enamel Formation; and, 4) Genetic Variation in Enamel Formation Genes.