The role of lead and cadmium in deciduous teeth and saliva on dental caries in children residing in Tehran, Iran.

BACKGROUND: Lead (Pb) and Cadmium (Cd) are important environmental contaminants. There is no biological monitoring of exposure to these heavy metals and their potential effect on dental caries in children in Tehran, Iran, a polluted megacity. Therefore, the present study investigated the potential association between Pb and Cd levels in primary teeth and saliva and dental caries. METHODS: In a cross-sectional design, 211 children aged 6-11 years referred to Tehran University of Medical Sciences, School of Dentistry and residing in Tehran were examined. Pb and Cd levels of exfoliated primary teeth and stimulated saliva were determined by atomic absorption spectrophotometry (AAS). Dental caries prevalence was evaluated according to WHO criteria. Socioeconomic status, oral hygiene behavior, snacking frequency and salivary pH data were acquired as confounding factors. Frequency and percentages were reported for categorical variables, mean and standard deviation (SD) for continuous variables, and geometric mean for skewed continuous variables. A simple linear regression and Pearson correlation tests were used for statistical analyses. P-values < 0.05 were considered as significant. RESULT: The mean (95 % confidence interval) Pb and Cd levels in teeth were 213.26 ppb (164.29-274.84) and 23.75 ppb (20.86-27.05), respectively. The mean Pb and Cd levels in saliva were 11.83 ppb (10.71-13.06) and 3.18 ppb (2.69-3.75), respectively. Furthermore, Pb and Cd in primary teeth and saliva were not associated (p > 0.05) with socioeconomic status, oral hygiene behavior and snacking frequency CONCLUSION: This study showed no association between Pb and Cd concentrations in primary teeth and in saliva with dental caries prevalence.

Longitudinal assessment of dental erosion-abrasion by cross-polarization optical coherence tomography in vitro

Abstract: This study tested a novel in vitro dental erosion-abrasion model and the performance of cross-polarization optical coherence tomography (CP-OCT) in longitudinally monitoring the simulated lesions. Thirty human enamel specimens were prepared and randomized to receive three dental erosion-abrasion (EA) protocols: severe (s-EA, lemon juice/pH:2.5/4.25%w/v citric acid), moderate (m-EA, grapefruit juice/pH:3.5/1.03%w/v citric acid) and no-EA (water, control). EA challenge was performed by exposing the specimens to acidic solutions 4x/day and to brushing 2x/day with 1:3 fluoridated toothpaste slurry, for 14 days. Enamel thickness measurements were obtained using CP-OCT at baseline (D0), 7 (D7) and 14 days (D14) and micro-computed tomography (micro-CT) at D14. Enamel surface loss was measured with both CP-OCT and optical profilometry at D0, D7 and D14. Data was analyzed with repeated-measures ANOVA and Pearson's correlation (r) (α = 0.05). CP-OCT enamel thickness decreased over time in the s-EA group (D0 >D7 > D14, p < 0.001) and m-EA group (D0 > D14, p = 0.019) but did not change in the no-EA group (p = 0.30). Overall, CP-OCT and micro-CT results at D14 correlated moderately (r = 0.73). CP-OCT surface loss was highest for s-EA (p <0.001) but did not differ between moderate and no-EA (p = 0.25). Enamel surface loss with profilometry increased with severity (no-EA>m-EA>s-EA, p < 0.001). D14 surface loss was higher than D7 for both methods except for the no-EA group with profilometry. CP-OCT and profilometry had moderate overall correlation (r = 0.70). Our results revealed that the currently proposed in vitro dental erosion-abrasion model is valid and could simulate lesions of different severities over time. CP-OCT was a suitable method for monitoring the EA lesions.

Effects of silver diamine fluoride on demineralization protection after a secondary acid challenge

Abstract Objective This investigation describes the effects of 5% sodium fluoride varnish and 38% silver diamine fluoride on demineralization protection of human enamel lesions of three different severities after a secondary acid challenge. Study design Specimens underwent color and enamel surface microhardness change measurements after demineralization and treatment events. Transverse microradiography was conducted following the secondary demineralization. Results After treatments, enamel surface microhardness change showed that 24-hour lesions treated with fluoride varnish had less rehardening than 24-hour lesions treated with silver diamine fluoride (p<0.05), whereas 144-hour lesions from both treatment groups showed a beneficial decrease in surface microhardness change that was markedly better in samples treated with silver diamine fluoride (p<0.05). After the secondary demineralization, 24- and 144-hour lesions treated with silver diamine fluoride showed a sustained beneficial decrease in enamel surface microhardness change when compared to fluoride varnish-treated samples of the corresponding lesion severity (p<0.05). Transverse microradiography showed no difference between fluoride varnish- and silver diamine fluoride-treated samples of any corresponding lesion severity, indicating that remineralization in both fluoride varnish- and silver diamine fluoride-treated samples was proportional to each other after a secondary acid challenge. Conclusions Using silver diamine fluoride may have comparable benefits to fluoride varnish in mineral loss prevention.

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.

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.

Caries lesion remineralization with fluoride toothpastes and chlorhexidine - effects of application timing and toothpaste surfactant.

INTRODUCTION: Habitual toothbrushing with fluoridated toothpaste followed by rinsing with antibacterial mouthwashes is a method to maintain good oral hygiene and to diminish the occurrence and severity of dental caries and periodontal disease. However, our understanding of how antimicrobial agents in mouthwashes affect fluoride-mediated caries lesion remineralization is still poor. OBJECTIVE: The objectives of this in vitro study were a) to determine the effects of the waiting period of chlorhexidine (CHX) rinsing after fluoride toothpaste use and b) to further determine the effect of the type of toothpaste surfactant [sodium dodecyl sulfate (SDS) or cocamidopropyl betaine (CAPB)] on caries lesion remineralization associated with CHX rinsing. MATERIAL AND METHODS: Caries lesions were formed in bovine enamel specimens and assigned to 10 treatment groups (n=18) based on Vickers surface microhardness (VHN). Lesions were then pH-cycled for 10 days with daily regimen comprised of twice daily toothpaste slurry treatments (1150 ppm fluoride, with SDS or CAPB), followed by CHX solution treatments [0, 15, 30 or 60 minutes following slurry treatment or no CHX treatment (negative control)]. VHN was measured again and the extent of lesion remineralization calculated (∆VHN). RESULTS: ∆VHN with SDS-toothpaste was significantly lower than with CAPB-toothpaste, indicating more remineralization for the CAPB-toothpaste. ∆VHN with 0-minute waiting time was significantly lower than with 30-minute waiting time and with negative control. CONCLUSIONS: The absence of CHX as an adjunct to fluoride toothpastes led to greater remineralization of enamel lesions compared with the immediate use of CHX treatment for both SDS- and CAPB-toothpastes. CAPB-toothpastes indicated significantly greater remineralization than SDS-toothpastes, and can be suggested for patients at high risk of caries. A 30-minute waiting time for CHX treatment is recommended after brushing.

Effects of a sodium fluoride- and phytate-containing dentifrice on remineralisation of enamel erosive lesions-an in situ randomised clinical study.

OBJECTIVE: The objective of this work was to evaluate effects of a dentifrice containing sodium fluoride (1150 ppm F) and the organic polyphosphate phytate (0.85% w/w of the hexa-sodium salt) on in situ remineralisation of early enamel erosive lesions and resistance to subsequent demineralisation. MATERIALS AND METHODS: Subjects (n = 62) wore palatal appliances holding eight bovine enamel specimens with pre-formed erosive lesions. They brushed their natural teeth with the phytate test dentifrice (TD); a positive control dentifrice (PC, 1150 ppm fluoride as NaF); a reference dentifrice (RD, disodium pyrophosphate + 1100 ppm fluoride as NaF) or a negative control dentifrice (NC, fluoride-free) in a randomised, double-blind, crossover design. Specimens were removed at 2, 4 and 8 h post-brushing and exposed to an ex vivo acid challenge. Surface microhardness (Knoop) was measured at each stage. The primary efficacy variable was relative erosion resistance (RER); other variables included the surface microhardness recovery (SMHR), acid resistance ratio (ARR) and enamel fluoride uptake (EFU). RESULTS: After 4 h, the results for RER, ARR and EFU were in the order PC > TD = RD > NC with PC > TD = RD = NC for SMHR. Results at 2 and 8 h were generally consistent with the 4 h data. Mineralisation progressed over time. Dentifrices were generally well-tolerated. CONCLUSIONS: In this in situ model, addition of phytate or pyrophosphate to a fluoride dentifrice inhibited the remineralising effect of fluoride. Both formulations still delivered fluoride to the enamel and inhibited demineralisation, albeit to a lesser extent than a polyphosphate-free dentifrice. CLINICAL RELEVANCE: Addition of phytate or pyrophosphate to a fluoride dentifrice may reduce its net anti-erosive properties.

Caries lesion remineralization with fluoride toothpastes and chlorhexidine - effects of application timing and toothpaste surfactant

Abstract Habitual toothbrushing with fluoridated toothpaste followed by rinsing with antibacterial mouthwashes is a method to maintain good oral hygiene and to diminish the occurrence and severity of dental caries and periodontal disease. However, our understanding of how antimicrobial agents in mouthwashes affect fluoride-mediated caries lesion remineralization is still poor. Objective: The objectives of this in vitro study were a) to determine the effects of the waiting period of chlorhexidine (CHX) rinsing after fluoride toothpaste use and b) to further determine the effect of the type of toothpaste surfactant [sodium dodecyl sulfate (SDS) or cocamidopropyl betaine (CAPB)] on caries lesion remineralization associated with CHX rinsing. Material and Methods: Caries lesions were formed in bovine enamel specimens and assigned to 10 treatment groups (n=18) based on Vickers surface microhardness (VHN). Lesions were then pH-cycled for 10 days with daily regimen comprised of twice daily toothpaste slurry treatments (1150 ppm fluoride, with SDS or CAPB), followed by CHX solution treatments [0, 15, 30 or 60 minutes following slurry treatment or no CHX treatment (negative control)]. VHN was measured again and the extent of lesion remineralization calculated (∆VHN). Results: ∆VHN with SDS-toothpaste was significantly lower than with CAPB-toothpaste, indicating more remineralization for the CAPB-toothpaste. ∆VHN with 0-minute waiting time was significantly lower than with 30-minute waiting time and with negative control. Conclusions: The absence of CHX as an adjunct to fluoride toothpastes led to greater remineralization of enamel lesions compared with the immediate use of CHX treatment for both SDS- and CAPB-toothpastes. CAPB-toothpastes indicated significantly greater remineralization than SDS-toothpastes, and can be suggested for patients at high risk of caries. A 30-minute waiting time for CHX treatment is recommended after brushing.

Influence of Toothbrushing on the Antierosive Effect of Film-Forming Agents.

This study evaluated the influence of toothbrushing on the antierosive effect of solutions containing sodium fluoride (225 ppm/F), stannous chloride (800 ppm/Sn), sodium linear polyphosphate (2%/LPP), and their combinations, and deionized water as negative control (C). Solutions were tested in a 5-day erosion-remineralization-abrasion cycling model, using enamel and dentin specimens (n = 8). Erosion was performed 6 times/day for 5 min, exposure to the test solutions 3 times/day for 2min, and toothbrushing (or not) with toothpaste slurry 2 times/day for 2 min (45 strokes). Surface loss (SL) was determined by noncontact profilometry. Data were analyzed using three-way ANOVA (α = 0.05). Brushing caused more SL than no brushing for enamel (mean ± SD, in micrometers: 52.7 ± 6.6 and 33.0 ± 4.5, respectively), but not for dentin (28.2 ± 1.9 and 26.6 ± 1.8, respectively). For enamel without brushing, F+LPP+Sn showed the lowest SL (23.8 ± 3.4), followed by F+Sn (30.6 ± 4.9) and F+LPP (31.7 ± 1.7), which did not differ from each other. No differences were found between the other groups and C (37.8 ± 2.1). When brushing, F+LPP+Sn exhibited the lowest SL (36.7 ± 2.4), not differing from F+LPP (39.1 ± 1.8). F, F+Sn and LPP+Sn were similar (46.7 ± 2.9, 42.1 ± 2.8 and 45.3 ± 4.6, respectively) and better than C (52.7 ± 4.3). Sn (55.0 ± 2.4) and LPP (51.0 ± 4.3) did not differ from C. For dentin, neither groups differed from C, regardless of brushing. In conclusion, toothbrushing did not affect the antierosive effect of F+Sn, F+LPP and F+LPP+Sn on enamel, although overall it led to more erosion than nonbrushing. F and LPP+Sn showed a protective effect only under brushing conditions, whereas Sn and LPP did not exhibit any protection. For dentin, neither toothbrushing nor the test solutions influenced the development of erosion.

The effect of fluoride varnishes on caries lesions: an in vitro investigation.

OBJECTIVES: The purpose of this study was to investigate the effect of five commercially available fluoride varnishes (FV) on caries lesions. MATERIALS AND METHODS: Ninety bovine enamel specimens were assigned to five varnish groups (n = 18). Early caries lesions were created in the specimens and characterized using Vickers surface microhardness number (VHN). FV was applied to each group of specimens. Immediately afterwards, 7.5 ml of artificial saliva (AS) were pipetted over each group of specimens, collected and renewed every 15 min for 6 h. AS samples were analyzed for fluoride using an ion-specific electrode. Enamel fluoride uptake (EFU) was determined using the acid etch technique. Each group was then subjected to a pH cycling regimen for 5 days after which VHN was determined again. One-way analysis of variance (ANOVA) was used for data analysis. RESULTS: FVs differed in their rehardening capability (highest mean value was for Enamel Pro = 32.3 ± 5.8 and lowest mean value was for Vanish = 18.9 ± 11.3). No significant difference in EFU was found among groups. Total fluoride release over 6 h was in the order of MI Varnish (303 µg/ml) > Enamel Pro (217 µg/ml) > Flor-Opal (153 µg/ml) > PreviDent(84 µg/ml) > Vanish(28 µg/ml). CONCLUSIONS: ΔVHN and fluoride release characteristics differ among FV products. These differences may be attributed to the different compositions and physical properties of the tested FV. CLINICAL RELEVANCE: Fundamental, comparable research on FV and how different formulations affect early caries lesion rehardening, fluoride release into saliva, and uptake by teeth is scarce.

Anti-erosive properties of solutions containing fluoride and different film-forming agents.

OBJECTIVES: To evaluate the anti-erosive potential of solutions containing sodium fluoride (NaF, 225 ppm F) and different film-forming agents. METHODS: In Phase 1, hydroxyapatite crystals were pre-treated with solutions containing NaF (F), linear sodium polyphosphate (LPP), sodium pyrophosphate tetrabasic (PP), sodium tripolyphosphate (STP), sodium caseinate (SC), bovine serum albumin (BSA), stannous chloride (Sn) and some combinations thereof. Deionized water was the control (C). The pH-stat method was used to evaluate hydroxyapatite dissolution. In Phase 2, the most effective solutions were tested in two independent experiments. Both consisted of an erosion-remineralization cycling model using enamel and dentine specimens with three solution treatments per day. In Phase 2a, the challenge was performed with 0.3% citric acid (pH=3.8). In Phase 2b, 1% citric acid (pH=2.4) was used. Hard tissue surface loss was determined profilometrically. Data were analyzed with two-way ANOVA and Tukey tests. RESULTS: In Phase 1, F, LPP, Sn and some of their combinations caused the greatest reduction in hydroxyapatite dissolution. In Phase 2a, C showed the highest enamel loss, followed by LPP. There were no differences between all other groups. In Phase 2b: (F+LPP+Sn) < (F+LPP) = (F+Sn) < (F) = (LPP+Sn) < (LPP) < (Sn) < C. For dentine, in both experiments, only the fluoride-containing groups showed lower surface loss than C, except for LPP+Sn in 2a. CONCLUSIONS: F, Sn, LPP reduced enamel erosion, this effect was enhanced by their combination under highly erosive conditions. For dentine, the F-containing groups showed similar protective effect. CLINICAL SIGNIFICANCE: The addition of LPP and/or Sn can improve the fluoride solution protection against erosion of enamel but not of dentine.

Efficacy of stannous, fluoride and their their combination in dentin erosion prevention in vitro

The aim of this study was to compare the protective effects of solutions containing stannous (Sn), fluoride (F) and their combination in the prevention of dentin erosion. Forty bovine root dentin specimens (4’4’2 mm3) were prepared and randomly assigned to 4 groups (n = 10): SnCl2(800 ppm/6.7 mM Sn), NaF (250 ppm/13 mM F), NaF/SnCl2 (800 ppm/6.7 mM Sn; 250 ppm/13 mM F), and deionized water (DIW) as a negative control. An acquired pellicle was formed on dentin samples by incubation in clarified, pooled, stimulated human saliva for 24 hours. The specimens were subjected to 5 daily cycles, each consisting of 5 of min demineralization (0.3%/15.6 mM citric acid, pH 2.6, 6’/day) and 60 min of re-mineralization in clarified human saliva. Thirty minutes after the 1st, 3rd and 5th demineralization episodes of each day, the specimens were treated with one of the test solutions for 2 min. Surface loss was measured via optical profilometry. Mixed-model ANOVA followed by Tukey’s test were used for the statistical analysis. Sn, F, and their combination significantly reduced the dentin surface loss by 23%, 36%, and 60% compared with DIW, respectively. All groups were significantly different (p < 0.05). The combination of Sn and F significantly reduced the amount of dentin surface loss compared with all other groups. The F group also significantly reduced surface loss compared with Sn and DIW, followed by the Sn group, which showed significantly greater protection compared with the DIW control. The daily use of a combined fluoride and stannous solution is promising for preventing dentin erosion.

A randomised clinical study to evaluate experimental children's toothpastes in an in-situ palatal caries model in children aged 11-14 years.

OBJECTIVES: To compare three children's sodium fluoride toothpastes to placebo with respect to enamel remineralisation potential, enamel fluoride uptake and net acid resistance using an in situ palatal caries model in children aged 11-14 years following a single brushing. DESIGN: This was a randomised, single blind (laboratory analyst), single-centre, four-treatment, crossover study with a 7-day washout period between treatments. The treatments were 1,426 ppm fluoride, 1,000 ppm fluoride, 500 ppm fluoride and 0 ppm fluoride (placebo) toothpaste (NaF/silica). A custom made in situ palatal appliance was used by each subject in all treatment periods. At each of the four treatment visits subjects wore the appliance containing four partially demineralised human enamel specimens for 5 minutes and then brushed their teeth using a standardised procedure for 60 seconds under supervision using 1.0 g (±0.1 g) of their assigned toothpaste. After 4 hours the appliance was removed and enamel specimen recovered. This process was repeated until all subjects completed all four study treatment visits. Recovered enamel specimens were analysed for per cent surface microhardness recovery (%SMHR; Knoop) and enamel fluoride uptake (EFU; microdrill biopsy). Subsequently, specimens were demineralised in vitro to determine their % net acid resistance (%NAR; Knoop). RESULTS: All three fluoride toothpastes demonstrated significantly greater %SMHR, EFU and %NAR compared with 0 ppm F toothpaste. The model demonstrated a dose response over the range 0 to 1,426 ppm fluoride for %SMHR, EFU and %NAR. There was no significant difference between 500 ppm F and 1,000 ppm F for %SMHR and between 1,000 ppm F and 1,426 ppm F for %SMHR, EFU and %NAR. CONCLUSIONS: The present in situ study demonstrated that the children's fluoride toothpastes tested are capable of delivering cariostatic amounts of fluoride to early caries lesions following a single brushing.

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.