Quantitative Proteomic Analysis of the Effect of Fluoride on the Acquired Enamel Pellicle

The acquired enamel pellicle (AEP) is a thin film formed by the selective adsorption of salivary proteins onto the enamel surface of teeth. The AEP forms a critical interface between the mineral phase of teeth (hydroxyapatite) and the oral microbial biofilm. This biofilm is the key feature responsible for the development of dental caries. Fluoride on enamel surface is well known to reduce caries by reducing the solubility of enamel to acid. Information on the effects of fluoride on AEP formation is limited. This study aimed to investigate the effects of fluoride treatment on hydroxyapatite on the subsequent formation of AEP. In addition, this study pioneered the use of label-free quantitative proteomics to better understand the composition of AEP proteins. Hydroxyapatite discs were randomly divided in 4 groups (n = 10 per group). Each disc was exposed to distilled water (control) or sodium fluoride solution (1, 2 or 5%) for 2 hours. Discs were then washed and immersed in human saliva for an additional 2 hours. AEP from each disc was collected and subjected to liquid chromatography electrospray ionization mass spectrometry for protein identification, characterization and quantification. A total of 45 proteins were present in all four groups, 12 proteins were exclusively present in the control group and another 19 proteins were only present in the discs treated with 5% sodium fluoride. Relative proteomic quantification was carried out for the 45 proteins observed in all four groups. Notably, the concentration of important salivary proteins, such as statherin and histatin 1, decrease with increasing levels of fluoride. It suggests that these proteins are repulsed when hydroxyapatite surface is coated with fluoride. Our data demonstrated that treatment of hydroxyapatite with fluoride (at high concentration) qualitatively and quantitatively modulates AEP formation, effects which in turn will likely impact the formation of oral biofilms.

Effect of NaF and TiF4 varnish and solution on bovine dentin erosion plus abrasion in vitro

Objectives. This in vitro study aimed to analyze the effect of TiF4 compared to NaF varnishes and solutions, to protect against dentin erosion associated with abrasion. Materials and methods. Bovine dentin specimens were pre-treated with NaF-Duraphat (2.26% F), NaF/CaF2-Duofluorid (5.63% F), experimental-NaF (2.45% F), experimental-TiF4 (2.45% F) and placebo varnishes; NaF (2.26% F) and TiF4 (2.45% F) solutions. Controls remained untreated. The erosive pH cycling was performed using a soft drink (pH 2.6) 4 x 90 s/day and the toothbrushing-abrasion 2 x 10 s/day, in vitro for 5 days. Between the challenges, the specimens were exposed to artificial saliva. Dentin tissue loss was measured profilometrically (mu m). Results. ANOVA/Tukey's test showed that all fluoridated varnishes (Duraphat, 7.5 +/- 1.1; Duofluorid, 6.8 +/- 1.1; NaF, 7.2 +/- 1.9; TiF4, 6.5 +/- 1.0) were able to significantly reduce dentin tissue loss (40.7% reduction compared to control) when compared to placebo varnish (11.2 +/- 1.3), control (11.8 +/- 1.7) and fluoridated (NaF, 9.9 +/- 1.8; TiF4, 10.3 +/- 2.1) solutions (p < 0.0001), which in turn did not significantly differ from each other. Conclusion. All fluoridated varnishes, but not the solutions, had a similar performance and a good potential to reduce dentin tissue loss under mild erosive and abrasive conditions in vitro. Risk patients for erosion and abrasion, especially those with exposed dentin, should benefit from this clinical preventive measure. Further research has to confirm this promising result in the clinical situation.

A comparative thermoluminescence and electron spin resonance study of synthetic carbonated A-type hydroxyapatite

Intensity of the 150 degrees C thermoluminescence peak of beta-irradiated carbonated synthetic A-type hydroxyapatite is approximately 12 times higher than that of the noncarbonated material. Deconvolution of the glow curve showed that this peak is a result of a trap distribution. An attempt was made to relate this thermoluminescence peak enhanced by carbonation with the ESR signal of the CO2- radical in natural or synthetic hydroxyapatite. (C) 2011 Elsevier Ltd. All rights reserved.

Combined effect of anterior malocclusion and inadequate lip coverage on dental trauma in primary teeth

Objectives: The main objective of this study was to investigate whether the interaction of malocclusion (open bite or increased overjet) combined with inadequate lip coverage strengthens its association with traumatic dental injury (TDI) in the primary teeth of preschool children compared to the presence of malocclusion alone. Subjects and methods: A cross-sectional survey was conducted with 376 children aged 3659 months who attended the National Day of Childrens Vaccination. Presence of TDI, tooth discoloration, and sinus tract were evaluated in the children. Variables associated with occlusion were also evaluated. A Poisson regression analysis was performed to verify the association between the explanatory variables and TDI as well as possible interactions among the variables. Then, the prevalence ratio was calculated. Results: The prevalence of TDI was 27.7%. The maxillary central incisor was the most affected tooth, without differences between the right and left sides. Boys had more dental trauma than girls (P = 0.04). The most common TDI was crown fracture restricted to the enamel (58.4%). Children with a combination of anterior open bite or increased overjet and inadequate lip coverage presented a higher prevalence of TDI than when the malocclusions were presented alone (P < 0.05). The same trends were observed when we included, in the final adjusted model, increased overjet instead of open bite. Conclusions: Anterior malocclusions of primary teeth such as increased overjet and anterior open bite are statistically significantly associated with dental trauma only when inadequate lip coverage is also present.

Cost-effectiveness of conventional and modified supervised toothbrushing in preventing caries in permanent molars among 5-year-old children

The cost-effectiveness of a modified supervised toothbrushing program was compared to a conventional program. A total of 284 five-year-old children presenting at least one permanent molar with emerged/sound occlusal surface participated. In the control group, oral health education and dental plaque dying followed by toothbrushing with fluoride dentifrice was carried outfour times per year. With the test group, children also underwent professional cross-brushing on surfaces of first permanent molar rendered by a dental assistant five times per year. Enamel/dentin caries were recorded on buccal, occlusal and lingual surfaces of permanent molars for a period of 18 months. The incidence density (ID) ratio was estimated using Poisson's regression model. The ID was 50% lower among boys in the test group (p = 0.016). The cost of the modified program was US$ 1.79 per capita. The marginal cost-effectiveness ratio among boys was US$ 6.30 per avoided carie. The modified supervised toothbrushing program was shown to be cost-effective in the case of boys.

Enzyme replacement prevents enamel defects in hypophosphatasia mice

Hypophosphatasia (HPP) is the inborn error of metabolism characterized by deficiency of alkaline phosphatase activity, leading to rickets or osteomalacia and to dental defects. HPP occurs from loss-of-function mutations within the gene that encodes the tissue-nonspecific isozyme of alkaline phosphatase (TNAP). TNAP knockout (Alpl-/-, aka Akp2-/-) mice closely phenocopy infantile HPP, including the rickets, vitamin B6-responsive seizures, improper dentin mineralization, and lack of acellular cementum. Here, we report that lack of TNAP in Alpl-/- mice also causes severe enamel defects, which are preventable by enzyme replacement with mineral-targeted TNAP (ENB-0040). Immunohistochemistry was used to map the spatiotemporal expression of TNAP in the tissues of the developing enamel organ of healthy mouse molars and incisors. We found strong, stage-specific expression of TNAP in ameloblasts. In the Alpl-/- mice, histological, mu CT, and scanning electron microscopy analysis showed reduced mineralization and disrupted organization of the rods and inter-rod structures in enamel of both the molars and incisors. All of these abnormalities were prevented in mice receiving from birth daily subcutaneous injections of mineral-targeting, human TNAP at 8.2?mg/kg/day for up to 44 days. These data reveal an important role for TNAP in enamel mineralization and demonstrate the efficacy of mineral-targeted TNAP to prevent enamel defects in HPP. (C) 2012 American Society for Bone and Mineral Research.

Performance of fluorescence-based and conventional methods of occlusal caries detection in primary molars - an in vitro study

International Journal of Paediatric Dentistry 2012; 22: 459466 Aim. This in vitro study aimed to test the performance of fluorescence-based methods in detecting occlusal caries lesions in primary molars compared to conventional methods. Design. Two examiners assessed 113 sites on 77 occlusal surfaces of primary molars using three fluorescence devices: DIAGNOdent (LF), DIAGNOdent pen (LFpen), and fluorescence camera (VistaProof-FC). Visual inspection (ICDAS) and radiographic methods were also evaluated. One examiner repeated the evaluations after one month. As reference standard method, the lesion depth was determined after sectioning and evaluation in stereomicroscope. The area under the ROC curve (Az), sensitivity, specificity, and accuracy of the methods were calculated at enamel (D1) and dentine caries (D3) lesions thresholds. The intra and interexaminer reproducibility were calculated using the intraclass correlation coefficient (ICC) and kappa statistics. Results. At D1, visual inspection presented higher sensitivities (0.970.99) but lower specificities (0.180.25). At D3, all the methods demonstrated similar performance (Az values around 0.90). Visual and radiographic methods showed a slightly higher specificity (values higher than 0.96) than the fluorescence based ones (values around 0.88). In general, all methods presented high reproducibility (ICC higher than 0.79). Conclusions. Although fluorescence-based and conventional methods present similar performance in detecting occlusal caries lesions in primary teeth, visual inspection alone seems to be sufficient to be used in clinical practice.

Calcium glycerophosphate supplemented to soft drinks reduces bovine enamel erosion

Objective: This in vitro study evaluated the effect of calcium glycerophosphate (CaGP) supplemented to soft drinks on bovine enamel erosion. Material and methods: Four pH-cycles were performed, alternating demineralization by the beverage and remineralization in artificial saliva. Results: Mean wear (+/- SD, mu m) was 7.91 +/- 1.13, 7.39 +/- 1.01, 7.50 +/- 0.91 and 5.21 +/- 1.08 for Coca-Cola (TM) without CaGP or containing CaGP at 0.1, 1.0 or 2.0 mM, respectively, while no wear was detected for CaGP at 5.0 and 10.0 mM. Corresponding figures for Sprite Zero (TM) without CaGP or containing CaGP at 0.1, 1.0, 2.0, 5.0 or 10.0 mM were 8.04 +/- 1.30, 7.84 +/- 0.71, 7.47 +/- 0.80, 4.96 +/- 0.81, 3.99 +/- 0.10 and 1.87 +/- 0.12, respectively. Conclusion: Supplementation of both beverages with CaGP seems to be an alternative to reduce their erosive potential.

Temperature rise during Er:YAG cavity preparation of primary enamel

This study aimed to assess in vitro thermal alterations taking place during the Er:YAG laser cavity preparation of primary tooth enamel at different energies and pulse repetition rates. Forty healthy human primary molars were bisected in a mesio-distal direction, thus providing 80 fragments. Two small orifices were made on the dentin surface to which type K thermocouples were attached. The fragments were individually fixed with wax in a cylindrical PlexiglassA (R) abutment and randomly assigned to eight groups, according to the laser parameters (n = 10): G1 -aEuro parts per thousand 250 mJ/ 3 Hz, G2 -aEuro parts per thousand 250 mJ/ 4 Hz, G3 -aEuro parts per thousand 250 mJ/ 6 Hz, G4 -aEuro parts per thousand 250 mJ/10 Hz, G5 -aEuro parts per thousand 250 mJ/ 15 Hz, G6 -aEuro parts per thousand 300 mJ/ 3 Hz, G7 -aEuro parts per thousand 300 mJ/ 4 Hz and G8 -aEuro parts per thousand 300 mJ/ 6 Hz. An area of 4 mm(2) was delimited. Cavities were done (2 mm long x 2 mm wide x 1 mm thick) using non-contact (12 mm) and focused mode. Temperature values were registered from the start of laser irradiation until the end of cavity preparation. Data were analyzed by one-way ANOVA and Tukey test (p a parts per thousand currency signaEuro parts per thousand 0.05). Groups G1, G2, G6, and G7 were statistically similar and furnished the lowest mean values of temperature rise. The set 250 mJ/10 and 15 Hz yielded the highest temperature values. The sets 250 and 300 mJ and 6 Hz provided temperatures with mean values below the acceptable critical value, suggesting that these parameters ablate the primary tooth enamel. Moreover, the temperature elevation was directly related to the increase in the employed pulse repetition rates. In addition, there was no direct correlation between temperature rise and energy density. Therefore, it is important to use a lower pulse frequency, such as 300 mJ and 6 Hz, during cavity preparation in pediatric patients.

Root and Root Canal Morphology of Four-rooted Maxillary Second Molars: A Micro-Computed Tomography Study

Introduction: This study examined the anatomy of 4-rooted maxillary second molars by using micro computed tomography. Methods: Twenty-five 4-rooted maxillary second molars were scanned to evaluate the size and curvature of the roots; the distance and spatial configuration between some anatomical landmarks; the number of root canals and the position of apical foramina; the occurrence of fusion of roots and enamel pearls; the configuration of the canal at the apical third; the cross-sectional appearance, the volume, and surface area of the root canals. Data were compared by using analysis of variance post hoc Tukey test (alpha = 0.05). Results: The specimens were classified as types I (n = 16), II (n = 7), and III (n = 2). The size of the roots was similar (P > .05), and most of them presented straight with 1 canal, except the mesiobuccal that showed 2 canals in 24% of the samples. The configuration of the pulp chamber was mostly irregular quadrilateral-shaped. The lowest mean distance of the orifices was observed between the buccal roots (P < .05). Accessory canals were present mostly in the apical third. Location of the apical foramina varied considerably. Fusion of roots and enamel pearls occurred in 44% and 8% of the samples, respectively. Mean distance from the pulp chamber floor to the furcation was 2.15 +/- 0.57 mm. No statistical differences were found in the bi-dimensional and 3-dimensional analyses (P > .05). Conclusions: All analyzed parameters showed differences between roots, except for the length of the roots, the configuration of the canals at the apical third, cross-sectional appearance, volume, and surface area of the canals. (J Endod 2012;38:977-982)

Degradation of the resin-dentin bonds after simulated and inhibited cariogenic challenge in an in situ model

The aim of this study was to evaluate the resindentin bonds of two simplified etch-and-rinse adhesive after simulated cariogenic and inhibited cariogenic challenge in situ. Dental cavities (4 mm wide, 4 mm long, and 1.5 mm deep) were prepared in 60 bovine teeth with enamel margins. Restorations were bonded with either adhesive Adper Single Bond 2 (3MESPE) or Optibond Solo Plus (Kerr). Forty restorations were included in an intra-oral palatal appliance that was used for 10 adult volunteers while the remaining 20 dental blocks were not submitted to any cariogenic challenge [NC group] and tested immediately. For the simulated cariogenic challenge [C+DA], each volunteer dropped 20% sucrose solution onto all blocks four times a day during 14 days and distilled water twice a day. In the inhibited cariogenic challenge group [C + FA], the same procedure was done, but slurry of fluoride dentifrice (1.100 ppm) was applied instead of water. The restored bovine blocks were sectioned to obtain a slice for cross-sectional Vickers microhardness evaluation and resindentin bonded sticks (0.8 mm2) for resindentin microtensile evaluation. Data were evaluated by two-way ANOVA and Tukey's tests (a = 0.05). Statistically lower microhardness values and degradation of the resindentin bonds were only found in the C + DW group for both adhesives. The in situ model seems to be a suitable short-term methodology to investigate the degradation of the resindentin bonds under a more realistic condition. (c) 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 100B: 14661471, 2012.

Screening of CO2 Laser (10.6 mu m) Parameters for Prevention of Enamel Erosion

Objective: The aim of this study was to screen CO2 laser (10.6 mu m) parameters to increase enamel resistance to a continuous-flow erosive challenge. Background data: A new clinical CO2 laser providing pulses of hundreds of microseconds, a range known to increase tooth acid-resistance, has been introduced in the market. Methods: Different laser parameters were tested in 12 groups (n = 20) with varying fluences from 0.1 to 0.9 J/cm(2), pulse durations from 80 to 400 mu s and repetition rates from 180 to 700 Hz. Non-lased samples (n = 30) served as controls. All samples were eroded by exposure to hydrochloric acid (pH 2.6) under continuous acid flow (60 mu L/min). Calcium and phosphate release into acid was monitored colorimetrically at 30 sec intervals up to 5 min and at 1 min intervals up to a total erosion time of 15 min. Scanning electron microscopic (SEM) analysis was performed in lased samples (n = 3). Data were statistically analysed by one-way ANOVA (p < 0.05) and Dunnett's post-hoc tests. Results: Calcium and phosphate release were significantly reduced by a maximum of 20% over time in samples irradiated with 0.4 J/cm(2) (200 mu s) at 450 Hz. Short-time reduction of calcium loss (<= 1.5 min) could be also achieved by irradiation with 0.7 J/cm(2) (300 mu s) at 200 and 300 Hz. Both parameters revealed surface modification. Conclusions: A set of CO2 laser parameters was found that could significantly reduce enamel mineral loss (20%) under in vitro erosive conditions. However, as all parameters also caused surface cracking, they are not recommended for clinical use.

Alveolar Bone Morphology in Patients With Bilateral Complete Cleft Lip and Palate in the Mixed Dentition: Cone Beam Computed Tomography Evaluation

Objectives: To verify the thickness and level of alveolar bone around the teeth adjacent to the cleft by means of cone beam computed tomography (CBCT) in patients with complete bilateral cleft lip and palate prior to bone graft surgery and orthodontic intervention. Method: The sample comprised 10 patients with complete bilateral cleft lip and palate (five boys and five girls) in the mixed dentition. The mean age was 9.5 years, and all subjects showed a G3 interarch relationship according to the Bauru index. The thickness of alveolar bone surrounding the maxillary incisors and the maxillary canines was measured in CBCT axial section using the software iCAT Xoran System. The distance between the alveolar bone crest and the cement-enamel junction (CEJ) was measured in cross sections. Results: The tomography images showed a thin alveolar bone plate around teeth adjacent to clefts. No bone dehiscence was observed in teeth adjacent to clefts during the mixed dentition. A slight increase in the distance between the alveolar bone crest and the CEJ was observed in the mesial and lingual aspects of canines adjacent to cleft. Conclusion: In patients with BCLP in the mixed dentition, teeth adjacent to the alveolar cleft are covered by a thin alveolar bone plate. However, the level of alveolar bone crest around these teeth seems to be normal, and no bone dehiscence was identified at this age.

In situ effect of chewing gum containing CPP–ACP on the mineral precipitation of eroded bovine enamel

Objectives: Stimulation of salivary flow is considered a preventive strategy for dental erosion. Alternatively, products containing calcium phosphate, such as a complex of casein phosphopeptide–amorphous calcium phosphate (CPP–ACP), have also been tested against dental erosion. Therefore, this in situ study analyzed the effect of chewing gum containing CPP–ACP on the mineral precipitation of initial bovine enamel erosion lesions. Methods: Twelve healthy adult subjects wore palatal appliances with two eroded bovine enamel samples. The erosion lesions were produced by immersion in 0.1% citric acid (pH 2.5) for 7 min. During three experimental crossover in situ phases (1 day each), the subjects chewed a type of gum, 3 times for 30 min, in each phase: with CPP–ACP (trident total), without CPP–ACP (trident), and no chewing gum (control). The Knoop surface microhardness was measured at baseline, after erosion in vitro and the mineral precipitation in situ. The differences in the degree of mineral precipitation were analyzed using repeated measures (RM-) ANOVA and post hoc Tukey’s test ( p < 0.05). Results: Significant differences were found among the remineralizing treatments ( p < 0.0001). Chewing gum (19% of microhardness recovery) improved the mineral precipitation compared to control (10%) and the addition of CPP–ACP into the gum promoted the best mineral precipitation effect (30%). Conclusions: Under this protocol, CPP–ACP chewing gum improved the mineral precipitation of eroded enamel. Clinical significance: Since the prevalence of dental erosion is steadily increasing, CPP–ACP chewing gum might be an important strategy to reduce th eprogression of initial erosion lesions.

Effect of Nd:YAG laser combined with fluoride on the prevention of primary tooth enamel demineralization

Most studies dealing with the caries preventive action of Nd:YAG laser have been done in permanent teeth and studies on primary teeth are still lacking. The aim of this study was to evaluate in vitro the effect of Nd:YAG laser combined or not with fluoride sources on the acid resistance of primary tooth enamel after artificial caries induction by assessing longitudinal microhardness and demineralization depth. Sixty enamel blocks obtained from the buccal/lingual surface of exfoliated human primary molars were coated with nail polish/wax, leaving only a 9 mm² area exposed on the outer enamel surface, and randomly assigned to 6 groups (n=10) according to the type of treatment: C-control (no treatment); APF: 1.23% acidulated phosphate fluoride gel; FV: 5% fluoride varnish; L: Nd:YAG laser 0.5 W/10 Hz in contact mode; APFL: fluoride gel + laser; FVL: fluoride varnish + laser. After treatment, the specimens were subjected to a des-remineralization cycle for induction of artificial caries lesions. Longitudinal microhardness data (%LMC) were analyzed by the Kruskal-Wallis test and demineralization depth data were analyzed by oneway ANOVA and Fisher’s LSD test (á=0.05). APFL and APF groups presented the lowest percentage of microhardness change (p<0.05). Demineralization depth was smaller in all treated groups compared with the untreated control. In conclusion, Nd:YAG laser combined or not with fluoride gel/varnish was not more effective than fluoride alone to prevent enamel demineralization within the experimental period.