Biomineralization of dentin.

A single biomineralization of demineralized dentin is significant to restore the demineralized dentin due to dental caries or erosion. In recent years, meaningful progress has been made regarding the mechanisms involved in the biomineralization of dentin collagen. Concepts changing from the classical ion-based crystallization to non-classical particle-based crystallization, inspired a different strategy to infiltrate the demineralized dentin collagen. The remarkable discovery was the report of liquid-like amorphous calcium phosphate as nanoprecursor particles to carbonated hydroxyapatite. The non-collagenous proteins and their analogues are widely investigated, for their key role in controlling mineralization during the process of crystal nucleation and growth. The in-depth studies of the gap zone provided significant improvements in our understanding of the structure of collagen and of the intrafibrillar remineralization of collagen fibrils. The collagen is not a passive substrate as previously supposed, and the active role of guiding nanoprecursor infiltration and mediating its nucleation has been demonstrated. Furthermore, recovery of mechanical properties has been evaluated to determine the effectiveness of dentin remineralization. Finally, the problems regarding the origin formation of the calcium phosphate that is deposited in the collagen, and the exact interactions between the non-collagenous proteins, amorphous calcium phosphate and collagen are still unclear. We reviewed the importance of these findings in enriching our understanding of dentin biomineralization, while addressing certain limitations that are inherent to in vitro studies.

Susceptibility of enamel to initial erosion in relation to tooth type, tooth surface and enamel depth.

This study aimed at assessing the susceptibility of different tooth types (molar/premolar), surfaces (buccal/lingual) and enamel depths (100, 200, 400 and 600 µm) to initial erosion measured by surface microhardness loss (ΔSMH) and calcium (Ca) release. Twenty molars and 20 premolars were divided into experimental and control groups, cut into lingual/ buccal halves, and ground/polished, removing 100 µm of enamel. The initial surface microhardness (SMH 0 ) was measured on all halves. The experimental group was subjected to 3 consecutive erosive challenges (30 ml/tooth of 1% citric acid, pH 3.6, 25 ° C, 1 min). After each challenge, ΔSMH and Ca release were measured. The same teeth were consecutively ground to 200, 400 and 600 µm depths, and the experimental group underwent 3 erosive challenges at each depth. No difference was found in SMH 0 between experimental and control groups. Multivariate nonparametric ANOVA showed no significant differences between lingual and buccal surfaces in ΔSMH (p = 0.801) or Ca release (p = 0.370). ΔSMH was significantly greater in premolars than in molars (p < 0.05), but not different with respect to enamel depth. Ca release decreased significantly with increasing depth. Regression between Ca release and ΔSMH at 100 µm depth showed lower slope and r 2 value, associated with greater Ca release values. At 200-600 µm depths, moderately large r 2 values were observed (0.651-0.830). In conclusion, different teeth and enamel depths have different susceptibility to erosion, so when Ca release is used to measure erosion, the depth of the test facet in enamel should be standardized, whereas this is less important if ΔSMH is used.

Sphingoid bases inhibit acid-induced demineralization of hydroxyapatite.

Calcium hydroxyapatite (HAp), the main constituent of dental enamel, is inherently susceptible to the etching and dissolving action of acids, resulting in tooth decay such as dental caries and dental erosion. Since the prevalence of erosive wear is gradually increasing, there is urgent need for agents that protect the enamel against erosive attacks. In the present study we studied in vitro the anti-erosive effects of a number of sphingolipids and sphingoid bases, which form the backbone of sphingolipids. Pretreatment of HAp discs with sphingosine, phytosphingosine (PHS), PHS phosphate and sphinganine significantly protected these against acid-induced demineralization by 80 ± 17%, 78 ± 17%, 78 ± 7% and 81 ± 8%, respectively (p < 0.001). On the other hand, sphingomyelin, acetyl PHS, octanoyl PHS and stearoyl PHS had no anti-erosive effects. Atomic force measurement revealed that HAp discs treated with PHS were almost completely and homogeneously covered by patches of PHS. This suggests that PHS and other sphingoid bases form layers on the surface of HAp, which act as diffusion barriers against H(+) ions. In principle, these anti-erosive properties make PHS and related sphingosines promising and attractive candidates as ingredients in oral care products.

Composition of enamel pellicle from dental erosion patients.

Oral health is dependent upon a thin mobile film of saliva on soft and hard tissues. Salivary proteins adhere to teeth to form the acquired enamel pellicle which is believed to protect teeth from acid erosion. This study investigated whether patients suffering diet-induced dental erosion had altered enamel pellicles. Thirty patients suffering erosion were compared to healthy age-matched controls. Subjects wore a maxillary splint holding hydroxyapatite and human enamel blocks for 1 h. The acquired enamel pellicle was removed from the blocks and compared to the natural incisor pellicle. Basic Erosive Wear Examination scores confirmed that dental erosion was present in erosion patients and absent from healthy age-matched controls. Erosion patients had half the amount of proteins (BCA assay) within the acquired pellicle forming on splint blocks compared to normal controls (p < 0.05). In particular, statherin, a calcium-binding protein, was 35% less abundant (p < 0.05). Calcium concentration within the acquired pellicle was also reduced by 50% in erosion patients (p < 0.001). In contrast, the natural pellicle on the incisor had similar amounts of total protein in erosion patients and healthy controls. In summary, the formation of new acquired pellicles on surfaces was reduced in erosion patients, which may explain their greater susceptibility to acid erosion of teeth.

Impact of laminar flow velocity of different acids on enamel calcium loss.

OBJECTIVE: The aim of the study was to evaluate the impact of flow velocity under laminar flow conditions of different acidic solutions on enamel erosion. MATERIAL AND METHODS: A total of 240 bovine enamel specimens were prepared and allocated to 30 groups (n = 8 each). Samples of 18 groups were superfused in a flow chamber system with laminar flow behavior using 1 ml of citric acid or hydrochloric acid (HCl) of pH 2.0, 2.6 or 3.0. Flow rates in the sample chamber were adjusted to 10, 60 or 100 µl/min. To simulate turbulent flow behavior, samples of six groups were immersed in 1 ml of the respective solution, which was vortexed (15 min, 600 rpm). For simulating non-agitated conditions, specimens of the remaining six groups were immersed in 1 ml of the respective solution without stirring. Calcium in the solutions, released from the enamel samples, was determined using Arsenazo III method. RESULTS: For acidic solutions of pH 2.6 and 3.0, erosive potential of citric acid was equivalent to that of HCl at a flow of 100 µl/min. The same observation was made for the samples subjected to turbulent conditions at pH 3. At all other conditions, citric acid induced a significantly higher calcium loss than HCl. CONCLUSION: It is concluded that under slow laminar flow conditions, flow rate variations lead to higher erosive impact of citric acid compared to hydrochloric acid at pH 2.0, but not at pH ≥ 2.6 and increasing laminar flow or turbulent conditions. CLINICAL RELEVANCE: Erosive enamel dissolution under laminar flow conditions is a complex issue influenced by flow rate and acidic substrate.

Novel bacterial proteolytic and metabolic activity associated with dental erosion-induced oral dysbiosis.

BACKGROUND: Dental erosion is a disease of the oral cavity where acids cause a loss of tooth enamel and is defined as having no bacterial involvement. The tooth surface is protected from acid attack by salivary proteins that make up the acquired enamel pellicle (AEP). Bacteria have been shown to readily degrade salivary proteins, and some of which are present in the AEP. This study aimed to explore the role of bacteria in dental erosion using a multi-omics approach by comparing saliva collected from participants with dental erosion and healthy controls. RESULTS: Salivary proteomics was assessed by liquid-chromatography mass spectrometry (LC-MS) and demonstrated two altered AEP proteins in erosion, prolactin inducible protein (PIP), and zinc-alpha-2 glycoprotein (ZAG). Immunoblotting further suggested that degradation of PIP and ZAG is associated with erosion. Salivary microbiome analysis was performed by sequencing the bacterial 16S rRNA gene (V1-V2 region, Illumina) and showed that participants with dental erosion had a significantly (p < 0.05) less diverse microbiome than healthy controls (observed and Shannon diversity). Sequencing of bacterial mRNA for gene expression (Illumina sequencing) demonstrated that genes over-expressed in saliva from erosion participants included H + proton transporter genes, and three protease genes (msrAB, vanY, and ppdC). Salivary metabolomics was assessed using nuclear magnetic resonance spectrometry (NMR). Metabolite concentrations correlated with gene expression, demonstrating that the dental erosion group had strong correlations between metabolites associated with protein degradation and amino acid fermentation. CONCLUSIONS: We conclude that microbial proteolysis of salivary proteins found in the protective acquired enamel pellicle strongly correlates with dental erosion, and we propose four novel microbial genes implicated in this process. Video Abstract.

The release of zinc, copper, lead, and cadmium from the mineral tissue of teeth under the influence of soft drinks and sour-tasting food.

This study was carried out with the aim of identifying the effects of consuming sour-tasting food and refreshing drinks on the bone tissue of teeth among teenagers. The cumulative effect of a year-long exposure of teeth to the erosive effects of a model system of acidic media (citric acid, lactic acid, acetic acid, apple vinegar, lemonade, the soft drink Sprite, mineral water) was studied. The effects were registered based on the amount of released biometal ions, of zinc and copper, and toxic lead, during a period of 24 hr at room temperature, using the potentiometric stripping analysis. In the given time span, amounts ranging from 75 to 750 ppm of zinc, from 0.1 to 1.0 ppm of copper, and up to 1.5 ppm of lead were released from the dental matrix, while the release of cadmium was below the level of detection. The changes to the mineral structure of the bone tissue were monitored by the Fourier's transformation infrared spectroscopy and scanning electron microscopy technique. These studies have shown that under the influence of an acidic medium significant erosion to the tooth enamel ensues and that the eroded surfaces had a radius of 1-5 µm.

Oral manifestations of gastroesophageal reflux disease.

Numerous case-control and other studies involving confirmation of gastroesophageal reflux disease (GERD) by esophageal pH-metry and the assessment of dental erosions have shown significant associations between the two conditions in both adults and children. By contrast, when asked to vote on whether GERD may cause dental erosions, only 42% of physicians strongly agreed that such an association existed in adults, and just 12.5% strongly agreed for children, respectively in two global consensus reports. Part of this divergence between the perceptions of physicians and the findings of research publications may reflect a general lack of oral health education during medical training, and cursory oral examinations being made under less-than-ideal conditions. Adequate salivary secretions are essential for the protection of the teeth and the oropharyngeal and esophageal mucosa. The quantity and quality of the saliva require monitoring as many drugs, including several of the proton pump inhibitors (PPIs), can cause hyposalivation. In addition, PPIs do not always result in adequate acid suppression. Therefore, collaboration between physicians and dentists is strongly advocated to prevent or ameliorate possible adverse oral effects from both endogenous and exogenous acids, and to promote adequate saliva production in patients with GERD.

Clinical study monitoring the pH on tooth surfaces in patients with and without erosion.

The aim of this study was to compare tooth surface pH after drinking orange juice or water in 39 patients with dental erosion and in 17 controls. The following investigations were carried out: measurement of pH values on selected tooth surfaces after ingestion of orange juice followed by ingestion of water (acid clearance), measurement of salivary flow rate and buffering capacity. Compared with the controls, patients with erosion showed significantly greater decreases in pH after drinking orange juice, and the pH stayed lower for a longer period of time (p < 0.05). Saliva parameters showed no significant differences between the two patient groups except for a lower buffering capacity at pH 5.5 in the erosion group.

Scanning electrochemical microscopy as a quantitative probe of acid-induced dissolution: theory and application to dental enamel.

This Article reports the use of scanning electrochemical microscopy (SECM) for the quantitative study of acid-induced dissolution. An ultramicroelectrode (UME) is used to generate a flux of protons galvanostatically just above a sample surface, creating controlled acid challenges relevant to acid erosion. The electrochemical technique produces etch features in the sample, which are characterized by white light interferometry (WLI). The technique has been applied to bovine enamel where understanding the kinetics of dissolution is important in the context of acid erosion. Dissolution has been observed as a fast process, but the high rates of mass transport in SECM allow the surface kinetics of dissolution to be evaluated. Key attributes of SECM for these studies are the ability to deliver high, controllable, and local acid challenges in a defined way and that multiple dissolution measurements can be performed on one sample, eliminating intersample variability effects. A novel moving boundary finite element model has been designed to describe the etching process, which allows the etch kinetics to be evaluated quantitatively, simply by measuring the size and shape of etch features over time.

Mechanism of action of tin-containing fluoride solutions as anti-erosive agents in dentine - an in vitro tin-uptake, tissue loss, and scanning electron microscopy study.

Solutions containing tin and fluoride exhibit remarkable anti-erosive properties with tin ions as a major agent. To elucidate its mechanism of action in dentine, the tin uptake on and in the tissue was investigated and related to histological findings and substance loss. Samples were treated twice daily, each treatment lasting for 2 min, with fluoride solutions [pH 4.5; 1,500 parts per million (p.p.m.) F] containing 2,100, 1,400, or 400 p.p.m. Sn as SnCl(2). In experiments 1 and 2, samples were eroded with citric acid (pH 2.3) six times each day, each treatment lasting for 5 min; in experiment 2, the demineralized organic matrix was continuously digested by collagenase; in experiment 3, no erosive challenges were performed. Sample surfaces and cross-sections were investigated using energy dispersive X-ray spectroscopy, scanning electron microscopy, and profilometry. Surface retention of tin was found in almost all treatment groups and was highest in experiment 2. On cross-sections, tin was retained within the organic matrix; in mineralized areas, tin was found mainly within a depth of 10 mum. Test solutions inhibited substance loss significantly; in experiment 2, the effect was dose-dependent. Erosion inhibition seemed to depend mainly on the incorporation of tin in the mineralized dentine when the organic portion was preserved, but on surface precipitation when the organic portion was continuously digested.

Effects of toothbrushing force on the mineral content and demineralized organic matrix of eroded dentine.

Eroded dentine has a complex histological structure, and its organic fraction becomes increasingly exposed as a result of the continual action of acids. The present study sought to investigate the effects of brushing forces up to 4 N on mineralized and organic dentine fractions. The study was a cyclic demineralization and remineralization experiment (carried out over 9 d). Erosion was performed with HCl (6 x 2 min d-1), pH 1.6. Samples exposed to erosion alone served as controls; test samples were eroded and brushed with a powered toothbrush (2 x 15 s d-1), applying forces of 2, 3, or 4 N. Samples were analysed (using profilometry and longitudinal microradiography) before and after the removal of superficial organic material with collagenase. Randomly selected samples were subjected to scanning electron microscopy. Demineralized organic material was present on all samples regardless of brushing force. Loss values (determined using profilometry) after erosion only, and after brushing with forces of 2, 3, and 4 N, were 11.7 +/- 5.1, 13.6 +/- 11.2, 30.7 +/- 19.0, and 25.5 +/- 20.3, respectively, before treatment with collagenase, and 111.7 +/- 11.6, 122.0 +/- 11.8, 121.9 +/- 15.7, and 123.0 +/- 12.0, respectively, after treatment with collagenase. Microradiography confirmed the results. Significant effects of brushing force were only found on the demineralized organic fraction, and mineral loss was unaffected. The notion that eroded dentine is particularly prone to abrasion should be reconsidered.

Comparison of calcium analysis, longitudinal microradiography and profilometry for the quantitative assessment of erosion in dentine.

Erosion of dentine causes mineral dissolution, while the organic compounds remain at the surface. Therefore, a determination of tissue loss is complicated. Established quantitative methods for the evaluation of enamel have also been used for dentine, but the suitability of these techniques in this field has not been systematically determined. Therefore, this study aimed to compare longitudinal microradiography (LMR), contacting (cPM) and non-contacting profilometry (ncPM), and analysis of dissolved calcium (Ca analysis) in the erosion solution. Results are discussed in the light of the histology of dentine erosion. Erosion was performed with 0.05 M citric acid (pH 2.5) for 30, 60, 90 or 120 min, and erosive loss was determined by each method. LMR, cPM and ncPM were performed before and after collagenase digestion of the demineralised organic surface layer, with an emphasis on moisture control. Scanning electron microscopy was performed on randomly selected specimens. All measurements were converted into micrometres. Profilometry was not suitable to adequately quantify mineral loss prior to collagenase digestion. After 120 min of erosion, values of 5.4 +/- 1.9 microm (ncPM) and 27.8 +/- 4.6 microm (cPM) were determined. Ca analysis revealed a mineral loss of 55.4 +/- 11.5 microm. The values for profilometry after matrix digestion were 43.0 +/- 5.5 microm (ncPM) and 46.9 +/- 6.2 (cPM). Relative and proportional biases were detected for all method comparisons. The mineral loss values were below the detection limit for LMR. The study revealed gross differences between methods, particularly when demineralised organic surface tissue was present. These results indicate that the choice of method is critical and depends on the parameter under study.

Fluoride penetration from toothpastes into incipient enamel erosive lesions investigated using dynamic secondary ion mass spectrometry.

OBJECTIVE: The primary aim of this study was to assess the utility of dynamic secondary ion mass spectrometry (DSIMS) as a convenient and sensitive technique for determining fluoride uptake and distribution into incipient human enamel erosive lesions in vitro. A secondary aim was to correlate the extent of lesion rehardening following treatment with a toothpaste slurry, with relative fluoride uptake determined by DSIMS. The final aim was to compare fluoride uptake by incipient lesions treated with toothpastes containing different sources of fluoride using DSIMS. METHODS: Relative fluoride uptake into the surface and body of enamel erosive lesions was monitored by DSIMS as a function of fluoride concentration in a series of formulation-matched experimental pastes. Fluoride uptake into lesions that had been subjected to treatment with different toothpaste slurries in a single-treatment enamel lesion rehardening model was also determined, and its relationship with regard to the extent of rehardening and also the fluoride source investigated. RESULTS: Fluoride uptake by incipient erosive lesions treated with toothpastes containing NaF was quantitatively compared by DSIMS and found to be directly proportional to the fluoride concentration over the studied range (0-1400 ppm). Lesion repair observed in a single-treatment lesion rehardening model was positively correlated with the extent of fluoride uptake by the treated lesions. DSIMS was also able to show differences between commercial toothpastes containing different sources of fluoride and their ability to deliver the fluoride into the body of the lesion. The detrimental effect of sodium hexametaphosphate (NaHMP) present in Crest Pro-Health formulations previously reported in the single-treatment lesion rehardening model was also evident from the DSIMS elemental line scans obtained from the lesion cross-sections. CONCLUSION: DSIMS has been shown to be a powerful selective technique for quantifying relative fluoride uptake into enamel erosive lesions, and determining the extent and depth of lesion penetration. The relative efficacy of toothpastes containing fluoride from a variety of sources in the single-treatment lesion rehardening study is positively correlated with fluoride uptake and penetration determined by DSIMS.

Effect of erosive and abrasive stress on sealing ability of different desensitizers: In-vitro study.

This in vitro study examined the sealing ability of different desensitizing agents under a chemo-mechanical stress condition. For the study, a total of 144 extracted, caries-free human third molars were used to produce 1 mm-thick dentin discs. The specimens were divided randomly into four groups: Superseal (SS), Gluma (GL), Gluma Self-etch (GS), and Tooth Coat (TC). For each group, the permeability was measured before and after applying the desensitizer, after being exposed to Coca Cola for 5 minutes, and after 3150 strokes of a brushing abrasion. The decrease in permeability after the erosive and abrasive stress was analyzed by ANOVA and Tukey post hoc test. As a result, the dentin permeability decreased significantly for all desensitizers immediately after application (p < 0.05). SS and GS showed a significant difference in permeability reduction observed immediately after application and after acid action with Coca Cola (p < 0.05). After brushing abrasion, the permeability reduction decreased significantly for all desensitizers tested in this study (p < 0.05). TC showed the largest decrease in dentinal permeability compared to that of the other desensitizers and the differences were significant after brushing abrasion (p < 0.05). All tested desensitizers were effective in reducing dentin permeability. The behavioral characteristics under erosive and abrasive stress varied according to the products used. TC exhibited excellent sealing ability among the other desensitizers.

Self-assembly of dental surface nanofilaments and remineralisation by SnF2 and CPP-ACP nanocomplexes.

Dental caries, erosion and hypersensitivity are major public health problems. SnF2 is used widely in oral care products to help prevent/treat these conditions. Casein phosphopeptide-stabilised amorphous calcium phosphate nanocomplexes (CPP-ACP) are a biomimetic nanotechnology of salivary phosphopeptide-ACP complexes that deliver bioavailable calcium and phosphate ions to promote dental remineralisation (repair). We show here using in vitro studies and a double-blind, randomised controlled, cross-over design in situ clinical trial that SnF2 and CPP-ACP interact to form a nanofilament coating on the tooth surface and that together they are superior in their ability to promote dental remineralisation. Sn(II) by cross-linking the CPP-ACP helps to stabilise the complexes which improves delivery to the tooth surface and enhances binding and ion incorporation into tooth mineral. The combination of SnF2 and CPP-ACP in oral care products may significantly improve their efficacy in prevention/treatment of dental caries/erosion and hypersensitivity.

Surface characteristics of a modified acidulated phosphate fluoride gel with nano-hydroxyapatite coating applied on bovine enamel subjected to an erosive environment.

This study evaluated the antierosive effect applying a modified acidulated phosphate fluoride (APF) gel containing nano-hydroxyapatite (nHAp) on the enamel surface before erosion. After polishing, the exposed flat enamel surfaces (n = 7/group) from bovine incisors were treated with artificial saliva (S - negative control), orange juice (ERO), APF gel (positive control) and APF_nHAp gel. All samples were subjected to six cycles of demineralization (orange juice, pH ~ 3.5, 10 min) followed by remineralization (saliva, 37°C, 1 hr). The enamel surface morphology, topography, and inorganic composition were analyzed using scanning electron microscopy, roughness testing, and micro energy-dispersive X-ray fluorescence spectrometry, respectively. The mean (standard deviation) roughness values (Ra, µm) were S, 0.13 (0.05); ERO, 0.25 (0.07); APF, 0.22 (0.08); and APF_nHAp, 0.17 (0.04). Ra values were significantly higher after ERO (p < .01) and APF (p < .05) treatments than after S. The enamel surface morphology was altered by the treatments, except for the S specimens. The mineral content of the enamel showed a clear trend with Ca and P reduction in the order of APF < S < APF_nHAp < ERO and APF < S < ERO < APF_nHAp, respectively. We can conclude that APF gel increased mineral concentration on the enamel. Moreover, the APF_nHAp material modified the composition and morphology of the enamel surface.

Retention of KOH-soluble fluoride on enamel and dentine under erosive conditions--A comparison of in vitro and in situ results.

OBJECTIVES: Aim of the study was to investigate the stability of CaF(2)-like precipitates on enamel and dentine under neutral or acidic conditions and to compare in vitro and in situ results. METHODS: Human enamel and dentine specimens were fluoridated (Elmex fluid, 5 min) and subjected to erosive demineralisation (Sprite light: 3/day each; 30 s in vitro, 2 min in situ) or stored under neutral conditions for 4 (in vitro) or 7 (in situ) days. KOH-soluble fluoride was determined using an ion selective electrode. Between the acid attacks, specimens were stored in remineralisation solution (in vitro) or retained in the oral cavity (four volunteers for enamel and dentine each). RESULTS: Initially, high amounts of KOH-soluble fluoride were gained (between 77.9+/-12.3 and 96.0+/-46.4 microg/cm2). Under neutral conditions in vitro, a significant decrease on enamel (16.2+/-5.0 microg/cm2) and dentine (18.6+/-10.5 microg/cm2) was found, which was more severe under acidic conditions (6.3+/-3.0 and 5.1+/-2.1 microg/cm2, respectively). Under in situ conditions, KOH-soluble fluoride was strikingly stable on enamel under neutral (42.3+/-12.6 microg/cm2) as well as under acidic conditions (54.1+/-17.4 microg/cm2). On dentine, the dissolution kinetics of KOH-soluble fluoride was similar to the in vitro conditions, but the loss was less severe (45.3+/-12.9 microg/cm2 under neutral and 8.8+/-6.4 microg/cm2 under acidic conditions). CONCLUSIONS: In vitro, more KOH-soluble fluoride was lost under erosive compared to neutral conditions. The intra-oral environment was considerably protective for CaF2-like precipitates especially on enamel.

Protective effect of the in situ pellicle on dentin erosion - an ex vivo pilot study.

AIM: The acquired pellicle is well known as an anti-erosive proteinaceous layer on enamel, but its protective properties on dentin have not been investigated in detail until now. The aim of the present ex vivo study was to evaluate the erosive effects on pellicle coated dentin. METHODS: Bovine dentin slabs were exposed to the oral cavity of one subject for 120 min for in situ pellicle formation. Subsequently, the slabs were incubated with HCl (pH 2.3) in vitro for 5 min and erosive calcium-release was measured photometrically. In addition, the acid treated specimens were evaluated by transmission electron microscopy (TEM). Pellicle free samples served as controls. RESULTS: Calcium erosion from the pellicle coated dentin slabs amounted to 23.5+/-2.9 microg Ca/min (pellicle free samples: 32.2+/-4.2 microg Ca/min). The difference was statistically significant (p < or = 0.05). In pellicle coated as well as in uncoated dentin samples, TEM-evaluation showed a demineralised dentinal surface layer which thickness ranged between 3 and 6 microm. The pellicle itself was partially dissolved but not removed by hydrochloric acid treatment. CONCLUSION: The protective properties of the acquired pellicle against an erosive challenge of the dentinal surface are limited. The dentinal pellicle functions like an ion permeable network rather than a barrier.

The impact of stannous, fluoride ions and its combination on enamel pellicle proteome and dental erosion prevention.

OBJECTIVES: To compare the effects of stannous (Sn) and fluoride (F) ions and their combination on acquired enamel pellicle (AEP) protein composition (proteome experiment), and protection against dental erosion (functional experiment). METHODS: In the proteome experiment, bovine enamel specimens were incubated in whole saliva supernatant for 24h for AEP formation. They were randomly assigned to 4 groups (n=10), according to the rinse treatment: Sn (800ppm/6.7mM, SnCl2), F (225ppm/13mM, NaF), Sn and F combination (Sn+F) and deionized water (DIW, negative control). The specimens were immersed 3× in the test rinses for 2min, 2h apart. Pellicles were collected, digested, and analyzed for protein content using liquid chromatography electrospray ionization tandem mass spectrometry. In the functional experiment, bovine enamel specimens (n=10) were similarly treated for pellicle formation. Then, they were subjected to a five-day erosion cycling model, consisting of 5min erosive challenges (15.6 mM citric acid, pH 2.6, 6×/d) and 2min treatment with the rinses containing Sn, F or Sn+F (3×/d). Between the treatments, all specimens were incubated in whole saliva supernatant. Surface loss was determined by profilometry. RESULTS: Our proteome approach on bovine enamel identified 72 proteins that were common to all groups. AEP of enamel treated with Sn+F demonstrated higher abundance for most of the identified proteins than the other groups. The functional experiment showed reduction of enamel surface loss for Sn+F (89%), Sn (67%) and F (42%) compared to DIW (all significantly different, p<0.05). CONCLUSION: This study highlighted that anti-erosion rinses (e.g. Sn+F) can modify quantitatively and qualitatively the AEP formed on bovine enamel. Moreover, our study demonstrated a combinatory effect that amplified the anti-erosive protection on tooth surface.