In vitro demineralization prevention by fluoride and silver nanoparticles when applied to sound enamel and enamel caries-like lesions of varying severities.

OBJECTIVES: To investigate the effect of fluoride and silver nanoparticles on the prevention of in vitro demineralization of sound enamel and enamel caries-like lesions of varying severities. METHODS: Caries-like lesions of different severities (1/6/15 days) were created in bovine enamel specimens. One group remained sound. All specimens were demineralized again using a partially saturated acetic acid solution. Mimicking the intra-oral retention of fluoride and silver in vitro, this solution was supplemented with fluoride (0/1/10 ppm) and/or silver nanoparticles (0/10 ppm) in a factorial design. Changes in lesion depth (ΔL) and integrated mineral loss (ΔΔZ) were evaluated by digital transverse microradiography. Data was analyzed using three-way ANOVA. RESULTS: Lesion severity significantly affected ΔΔZ and ΔL, after no treatment and after the treatment of fluoride and silver independently (p = 0.012 and p = 0.037, respectively). Fluoride and the fluoride × lesion severity interaction were shown to be significant (p < 0.001) on ΔΔZ and ΔL. Silver nanoparticles significantly affected ΔΔZ (p = 0.041), but not ΔL (p = 0.15). The silver nanoparticles × lesion severity interaction was significant for ΔΔZ and ΔL (p = 0.032 and p = 0.024, respectively). No interaction was observed for ΔΔZ and ΔL between fluoride and silver (p = 0.962 and p = 0.971, respectively) as well as lesion severity and the use of fluoride and silver combined (p = 0.722 and p = 0.158, respectively). CONCLUSION: Fluoride and silver nanoparticles had a significant effect on the prevention of in vitro demineralization of sound enamel and enamel caries-like lesions of varying severities. CLINICAL SIGNIFICANCE: Fluoride and silver nanoparticles may potentially allow for more tailored caries prevention.

A visual evaluation of oral plaque removal utilizing an adjunct enzyme pre-rinse in orthodontic subjects.

OBJECTIVE: To determine if an adjunct proteolytic pre-rinse along with contemporary methods of dental cleaning may more effectively remove visual plaque in subjects with fixed orthodontic appliances. MATERIALS AND METHODS: Forty-three orthodontic subjects, ages 10 to 25, completed this single site, double-blind, crossover clinical trial. Subjects randomly received bromelain enzyme or a powdered-sugar placebo pre-rinse, followed by manual tooth brushing and use of a Waterpik. Subjects received the alternate pre-rinse during the subsequent visit. Baseline and residual plaque accumulation were recorded via disclosing tablet and digital photography. A single, blinded examiner scored visual plaque scores from randomized photographs. Treatment effects on composite plaque score were evaluated using repeated-measures analysis of variance. A 5% significance level was used for all tests. RESULTS: No significant differences in plaque scores were noted at baseline or post-rinse between the enzyme and placebo. The changes from baseline to post-rinse (P = .190), post-brushing (P = .764), and post-Waterpik (P = .882) were not significantly different between interventions. Significant reduction in plaque scores were observed in both arms of the study after brushing (P < .01) and waterjet use (P < .01). Neither age (P = .220) nor gender (P = .449) impacted plaque scores. CONCLUSIONS: Use of a bromelain enzyme pre-rinse alone did not significantly enhance plaque removal. A significant reduction in retained plaque was observed with the application of brushing and or Waterpik.

Effects of casein phosphopeptide-amorphous calcium phosphate crème on nicotine-induced Streptococcus mutans biofilm in vitro.

OBJECTIVES: The aim of this study was to test the effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) crème, or MI Paste™ (MIP), on nicotine-induced Streptococcus mutans biofilm. The experiment utilized S. mutans biofilm assays with varying concentrations of nicotine and MIP aqueous concentrate levels. First hand exposure to nicotine has been demonstrated to significantly increase S. mutans biofilm formation, while the active component, CPP-ACP, in MIP has been shown to reduce S. mutans biofilm formation. MATERIALS AND METHODS: A 24-h culture of S. mutans UA159 in microtiter plates were treated with varying nicotine concentrations (0-32 mg/ml) in Tryptic Soy Broth supplemented with 1% sucrose (TSBS) with or without MIP aqueous concentrate. A spectrophotometer was used to determine total growth absorbance and planktonic growth. The microtiter plate wells were washed, fixed, and stained with crystal violet dye and the absorbance measured to determine biofilm formation. RESULTS: The presence of MIP aqueous concentrate inhibits nicotine-induced S. mutans biofilm formation at different concentrations of nicotine (0-32 mg/ml). CONCLUSION: The results demonstrated nicotine-induced S. mutans biofilm formation is decreased in the presence of MIP. This provides further evidence about the cariostatic properties of CPP-ACP, the active soluble ingredient in the MIP, and reconfirms the harmful effects of nicotine. CLINICAL SIGNIFICANCE: Smokers may gain dual benefits from the use of MIP, as a remineralization agent and as a cariostatic agent, by inhibiting nicotine-induced S. mutans biofilm formation.

Effectiveness of in vitro primary coronal caries prevention with silver diamine fluoride - Chemical vs biofilm models.

OBJECTIVES: The main goal of this study was to investigate the effectiveness of SDF and its individual components, silver (Ag+) and fluoride (F-) ions, in preventing enamel demineralization using biofilm and chemical models. METHODES: Polished human enamel specimens were assigned to five treatment groups (n = 18 per group): SDF (38 %); SDF followed by application of a saturated solution of potassium iodide (SDF + KI); silver nitrate (AgNO3; silver control, 253,900 ppm Ag+); potassium fluoride (KF; fluoride control, 44,800 ppm F); deionized water (DIW). Treatments were applied once to sound enamel. In the biofilm model, specimens were demineralized by aerobic overnight incubation using cariogenic bacteria isolated from human saliva in brain heart infusion supplemented with 0.2 % sucrose for three days. In the chemical model, enamel specimens were immersed in a demineralizing solution containing 0.1 M lactic acid, 4.1 mM CaCl2, 8.0 mM KH2PO4, 0.2 % Carbopol 907, pH adjusted to 5.0 for five days. Vickers surface microhardness was used to determine the extent of enamel demineralization. Data were analyzed using one-way ANOVA. RESULTS: In the chemical model, there was no statistically significant difference between SDF and SDF + KI in preventing coronal caries (p < 0.0001). In the biofilm model, SDF + KI was significantly less effective in preventing demineralization than SDF (p < 0.0001). In both models, SDF and SDF + KI were superior in their ability to prevent caries lesion formation than AgNO3 and DIW. CONCLUSION: KI application after SDF treatment appears to impair SDF's ability to prevent biofilm-mediated but not chemically induced demineralization. CLINICAL SIGNIFICANCE: SDF may be a viable option in preventing primary coronal caries.

Influence of salivary conditioning and sucrose concentration on biofilm-mediated enamel demineralization.

INTRODUCTION: The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types. OBJECTIVE: To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization. METHODOLOGY: Saliva collection was approved by Indiana University IRB. Three donors provided wax-stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose-supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA. RESULTS: The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001). CONCLUSION: Within the study limitations, surface conditioning using human saliva does not influence biofilm-mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm-mediated demineralization of the enamel surface.

Chapter 3: Macroelements: Ca, Na, K, P, Cl.

The foods in the diet contain a wide range of organic and inorganic compounds. Considering these from an elemental perspective, 5 so-called macroelements, calcium, potassium, sodium, phosphorus and chlorine, are contained in comparatively large quantities in foods compared to all other elements. This chapter attempts to review the importance of these dietary macroelements on oral health, and in particular their role in tooth loss, dental caries, erosive tooth wear and periodontal disease. Calcium and phosphate make up the bulk of the mineralized human tissues. Adequate intake of both is therefore of crucial importance in maintaining the health, function and retention of teeth and bones. Supplementation of the diet with calcium has also been shown to aid in maintaining and improving oral health. Several attempts have been made to lessen the erosive potential of beverages through calcium supplementation. Adequate calcium intake is also crucial for maintaining periodontal health. In many areas, however, the evidence is still emerging or controversial. Phosphate supplementation of the diet was once thought to decrease caries incidence, although studies in children were not successful. Furthermore, little attention has been paid to the other macroelements, highlighting the need for more well-controlled and comprehensive studies.

Influence of salivary conditioning and sucrose concentration on biofilm-mediated enamel demineralization

Abstract The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types. Objective To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization. Methodology Saliva collection was approved by Indiana University IRB. Three donors provided wax-stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose-supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA. Results The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001). Conclusion Within the study limitations, surface conditioning using human saliva does not influence biofilm-mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm-mediated demineralization of the enamel surface.

Photoinhibition of Streptococcus mutans Biofilm-Induced Lesions in Human Dentin by Violet-Blue Light.

This in vitro study determined the effectiveness of violet-blue light on Streptococcus mutans (UA159) biofilm induced dentinal lesions. Biofilm was formed on human dentin specimens in a 96-well microtiter plate and incubated for 13 h in the presence of tryptic soy broth (TSB) or TSB supplemented with 1% sucrose (TSBS). Violet-blue light (405 nm) from quantitative light-induced fluorescence (QLFTM) was used to irradiate the biofilm. Supernatant liquid was removed, and the biofilm was irradiated continuously with QLF for 5 min twice daily with an interval of 6 h for 5 d, except with one treatment on the final day. Colony forming units (CFU) of the treated biofilm, changes in fluorescence (∆F; QLF-Digital BiluminatorTM), lesion depth (L), and integrated mineral loss (∆Z; both transverse microradiography) were quantified at the end of the fifth day. Statistical analysis used analysis of variance (ANOVA), testing at a 5% significance level. In the violet-blue light irradiated groups, there was a significant reduction (p < 0.05) of bacterial viability (CFU) of S. mutans with TSB and TSBS. Violet-blue light irradiation resulted in the reduction of ∆F and L of the dentinal surface with TSBS. These results indicate that violet-blue light has the capacity to reduce S. mutans cell numbers.

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.

Erosion protection by calcium lactate/sodium fluoride rinses under different salivary flows in vitro.

This study investigated the effect of a calcium lactate prerinse on sodium fluoride protection in an in vitro erosion-remineralization model simulating two different salivary flow rates. Enamel and dentin specimens were randomly assigned to 6 groups (n = 8), according to the combination between rinse treatments - deionized water (DIW), 12 mM NaF (NaF) or 150 mM calcium lactate followed by NaF (CaL + NaF) - and unstimulated salivary flow rates - 0.5 or 0.05 ml/min - simulating normal and low salivary flow rates, respectively. The specimens were placed into custom-made devices, creating a sealed chamber on the specimen surface connected to a peristaltic pump. Citric acid was injected into the chamber for 2 min, followed by artificial saliva (0.5 or 0.05 ml/min) for 60 min. This cycle was repeated 4×/day for 3 days. Rinse treatments were performed daily 30 min after the 1st and 4th erosive challenges, for 1 min each time. Surface loss was determined by optical profilometry. KOH-soluble fluoride and structurally bound fluoride were determined in specimens at the end of the experiment. Data were analyzed by 2-way ANOVA and Tukey tests (α = 0.05). NaF and CaL + NaF exhibited significantly lower enamel and dentin loss than DIW, with no difference between them for normal flow conditions. The low salivary flow rate increased enamel and dentin loss, except for CaL + NaF, which presented overall higher KOH-soluble and structurally bound fluoride levels. The results suggest that the NaF rinse was able to reduce erosion progression. Although the CaL prerinse considerably increased F availability, it enhanced NaF protection against dentin erosion only under hyposalivatory conditions.

The effect of strontium and combinations of strontium and fluoride on the remineralization of artificial caries lesions in vitro.

OBJECTIVE: To determine the effect of relatively low strontium concentrations on enamel remineralization and investigate the dose-response effects of strontium and fluoride combinations on the remineralization of artificial caries lesions in vitro. METHOD AND MATERIALS: Artificial caries lesions were created in 135 bovine enamel specimens. Lesion severity was analyzed using transverse microradiography (TMR) and quantitative light-induced fluorescence (QLF). The specimens were randomly assigned to nine treatment groups based on lesion volume after lesion creation, as measured by TMR. Treatment groups were based on a 3 x 3 factorial design (0/0.05/0.1 ppm fluoride and 0/10/15 ppm strontium). Lesions were remineralized at 37°C for 14 days in artificial saliva, which was supplemented or not with NaF and/or SrCl2 x 6H2O. Lesion remineralization was assessed using QLF and TMR. Data were analyzed using ANOVA. RESULTS: For the TMR data, lesion remineralization in the 10 ppm strontium + 0.05 ppm fluoride group was significantly higher than in all other groups (P < .05) except the 0 ppm strontium + 0.05 ppm fluoride group (P = .06). The 10 ppm strontium + 0 ppm fluoride group exhibited significantly less remineralization than the 0 ppm strontium + 0 ppm fluoride group (P = .048). For the QLF data, intergroup differences were not the same as for the TMR analysis. The QLF measurement was only moderately correlated with TMR mineral loss (r = -0.37). CONCLUSION: Strontium alone did not improve the remineralization of artificial caries lesions under the chosen in vitro conditions. However, a synergistic effect between the combination of fluoride and strontium was found at specific concentrations.