Addition of Grape Seed Extract Renders Phosphoric Acid a Collagen-stabilizing Etchant.

Previous studies found that grape seed extract (GSE), which is rich in proanthocyanidins, could protect demineralized dentin collagen from collagenolytic activities following clinically relevant treatment. Because of proanthocyanidin's adverse interference to resin polymerization, it was believed that GSE should be applied and then rinsed off in a separate step, which in effect increases the complexity of the bonding procedure. The present study aimed to investigate the feasibility of combining GSE treatment with phosphoric acid etching to address the issue. It is also the first attempt to formulate collagen-cross-linking dental etchants. Based on Fourier-transformed infrared spectroscopy and digestion assay, it was established that in the presence of 20% to 5% phosphoric acid, 30 sec of GSE treatment rendered demineralized dentin collagen inert to bacterial collagenase digestion. Based on this positive result, the simultaneous dentin etching and collagen protecting of GSE-containing phosphoric acid was evaluated on the premise of a 30-second etching time. According to micro-Raman spectroscopy, the formulation containing 20% phosphoric acid was found to lead to overetching. Based on scanning and transmission electronic microscopy, this same formulation exhibited unsynchronized phosphoric acid and GSE penetration. Therefore, addition of GSE did render phosphoric acid a collagen-stabilizing etchant, but the preferable phosphoric acid concentration should be <20%.

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

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

Effects of fluoride on in vitro enamel demineralization analyzed by ¹9F MAS-NMR.

The mechanistic action of fluoride on inhibition of enamel demineralization was investigated using (19)F magic angle spinning nuclear magnetic resonance (MAS-NMR). The aim of this study was to monitor the fluoride-mineral phase formed on the enamel as a function of the concentration of fluoride ions [F(-)] in the demineralizing medium. The secondary aim was to investigate fluorapatite formation on enamel in the mechanism of fluoride anti-caries efficacy. Enamel blocks were immersed into demineralization solutions of 0.1 M acetic acid (pH 4) with increasing concentrations of fluoride up to 2,262 ppm. At and below 45 ppm [F(-)] in the solution, (19)F MAS-NMR showed fluoride-substituted apatite formation, and above 45 ppm, calcium fluoride (CaF2) formed in increasing proportions. Further increases in [F(-)] caused no further reduction in demineralization, but increased the proportion of CaF2 formed. Additionally, the combined effect of strontium and fluoride on enamel demineralization was also investigated using (19)F MAS-NMR. The presence of 43 ppm [Sr(2+)] in addition to 45 ppm [F(-)] increases the fraction of fluoride-substituted apatite, but delays formation of CaF2 when compared to the demineralization of enamel in fluoride-only solution.

Dentin decalcification during lithium treatment: case report.

Severe dental decay and changes in tooth structure have been reported in association with the use of lithium in Psychiatry, but lithium effects on tooth inorganic composition remain unknown. A 30-year-old woman with bipolar disorder, treated with lithium carbonate presented severe dental decay. Dentin samples from lithium and healthy volunteers were collected and submitted to ionic and ultrastructural analysis. Samples from the lithium patient exhibited irregular peritubular walls and the mineral crystals were irregularly arranged in the intertubular dentin. In addition, a decrease in Mg/P/Ca and an increase of Zn concentrations were detected. These data suggest that the severe dental decay and changes in the tooth structure observed for the lithium-treated patient are related to dentin mineral loss and that this pathological condition is different from caries lesions.

Subablative Er:YAG laser effect on enamel demineralization.

OBJECTIVES: To characterize the cariostatic potential of a low-energy Er:YAG laser treatment. METHODS: Twelve sound premolars were selected. Two 2 × 1 mm windows were created on each tooth and randomly assigned to L(1) and L(2) groups. Three sites in each window were chosen with the middle site as the control and the left and right ones receiving Er:YAG laser treatment of 5.1 J/cm(2) (L(1)) or 2.0 J/cm(2) (L(2)), respectively. The teeth were further subjected to 4-day pH cycling to create caries-like lesions. After mineral quantification using a micro-computed tomography scanner, the preventive effects (ΔML = mineral loss of the control area minus that of the lased area) of L(1) and L(2) treatments were calculated based on the difference in the gray value of the control and lased sites. RESULTS: Significant inhibitory effects of L(1) and L(2) on enamel demineralization were demonstrated (both p ≤ 0.001), with the L(1) treatment having a greater effect (45.2%) than the L(2) treatment (25.2%, p = 0.004). CONCLUSIONS: Subablative low-energy Er:YAG laser irradiation can significantly prevent enamel demineralization potentially through the retardation of enamel diffusion. This study confirmed that high-energy laser treatment, which may damage the peripheral and underlying tissues, may not be needed for caries prevention.

Mechanical and micromorphological evaluation of chlorhexidine-mediated dentin remineralization.

Chlorhexidine (CHX) has been reported to reduce self-degradation of collagen fibrils by inhibiting host-derived protease activity in demineralized dentin. Theoretically, if the collagen fibril scaffold of demineralized dentin maintains its original crosslinkage pattern on treatment with CHX and appropriate supplementation with necessary mineral sources, dentin remineralization may occur in demineralized lesions. In this study, we provide direct mechanical and micromorphological evidence for the ability of CHX to promote remineralization of demineralized dentin. Specifically, with respect to demineralized dentin blocks treated with different concentrations of CHX (0.02-2%) and stored in simulated body fluid, we have observed a significant increase in the elastic modulus of dentin treated with relatively high concentrations of CHX (0.2 and 2%) as storage time increased, whereas the elastic modulus of the non-CHX treated control group decreased. We have also observed a dense mineral deposition along collagen fibrils in the dentin group treated with 0.2 and 2% CHX via field emission scanning electron microscopy.

An in vitro model for the study of chemical exchange between glass ionomer restorations and partially demineralized dentin using a minimally invasive restorative technique.

OBJECTIVES: This study was designed to validate an in vitro model of the Atraumatic Restorative Technique (ART). This model allowed in depth analyses of the migration of apatite forming Sr and F, from a glass-ionomer, Fuji IXGP (GC Corporation, Tokyo, Japan), into partly demineralized dentine. The second objective was to study the effects of the levels of mineral loss on such ionic exchange. METHODS: Artificial lesions were created in thirty third molars, which were divided into three groups based on time of exposure to the demineralizing solution (7, 14 and 21 days). These were restored with Fuji IXGP and left for 21 days. The method of analysis was electron probe microanalysis (EPMA) with wavelength dispersive spectrometry (WDS). This combination can yield both qualitative identification of elements as well as quantitative compositional information, with a minimum detection limit of 0.01%. RESULTS: The results indicated that there was no significant difference in the levels of Ca and P from the control and test sides for all three groups of demineralized dentine. The differences in the depth of penetration and the amount of Sr and F between control and test were significant. CONCLUSIONS: The validity of the in vitro model was confirmed. There was migration of both strontium and fluorine to the artificially demineralized dentine in a pattern consistent with remineralization and similar to what was observed in a previous in vivo study.

Morphological and chemical characterization of tooth enamel exposed to alkaline agents.

OBJECTIVES: In this study, morphological and chemical changes in teeth enamel exposed to alkaline agents, with or without surfactants, have been investigated. In addition, chemical effects of the organic surface layer, i.e. plaque and pellicle, were also investigated. METHODS: The present study was conducted using several techniques: Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD) and scanning electron microscopy (SEM). RESULTS: From XPS-measurements, it was found that exposure to alkaline solutions resulted in a massive removal of carbon from the tooth surface, and that the addition of surfactants increased the rate under present conditions. Based on the results from the FTIR-analysis, no substitution reactions between phosphate, carbonate and hydroxide ions in the enamel apatite could be detected. From a minor SEM-analysis, degradation and loss of substance of the enamel surface was found for the exposed samples. From XRD-analysis, no changes in crystallinity of the enamel apatite could be found between the samples. CONCLUSIONS: The findings in this study show that exposure to alkaline solutions results in a degradation of enamel surfaces very dissimilar from acidic erosion. No significant erosion or chemical substitution of the apatite crystals themselves could be discerned. However, significant loss of organic carbon at the enamel surface was found in all exposed samples. The degradation of the protective organic layer at the enamel surface may profoundly increase the risk for caries and dental erosion from acidic foods and beverages.

Degree of mineral loss in softened human enamel after acid erosion measured by chemical analysis.

OBJECTIVES: Erosive attack on enamel leads to a bulk material loss and a partial demineralisation at the lesion body. This zone of softened enamel has a reduced physical stability and large interprismatic porosities. The aim of the current study was to measure the degree of demineralisation with respect to the content of calcium and inorganic phosphorus in softened enamel using ultrasonication and chemical analysis. The erosive demineralisation was to be compared with demineralisation in caries. METHODS: Erosion of 10 polished human enamel samples was performed in 0.3% citric acid adjusted to pH 3.2 at 35 degrees C in a slowly stirred solution, each in a volume of 15 ml. After erosion the softened enamel was removed by ultrasonication in 2 ml of deionised water and enamel crystals were dissolved by adding 220 microl of 1M HCl. Substance loss was measured profilometricaly after erosion and after ultrasonication. Surface size of eroded enamel was measured thus volume loss could be calculated. Calcium and phosphate ion concentration in the solutions were measured after ultrasonication. RESULTS: Citric acid erosion caused a mean substance loss of 16.0 microm (SD 2.5 microm) and further enamel softening of 2.4 microm (SD 1.5 microm). The degree of demineralisation of softened enamel, in comparison the original mineral content, was 62% for calcium and 64% for inorganic phosphorus was calculated. CONCLUSION: The high degree of demineralisation shown in the current study explains the instability of softened enamel even against mild physical impact.

Effect of a calcium glycerophosphate fluoride dentifrice formulation on enamel demineralization in situ.

PURPOSE: To evaluate in situ the effect and mechanisms involved in the anticariogenic effect of a calcium glycerophosphate fluoride dentifrice. METHODS: In a double-blind, crossover design, a non-F dentifrice (negative control), a F dentifrice and a F dentifrice containing 0.13% CaGP were compared regarding the inhibition of enamel demineralization. Both F dentifrices contained 1500 microg F/g (w/w) as sodium monofluorophosphate (MFP). Bovine enamel blocks were mounted in contact with a S. mutans test plaque, in palatal appliances worn by 10 volunteers. 30 minutes after treatment with the dentifrices, a sucrose rinse was performed and enamel demineralization was assessed after an additional 45 minutes. RESULTS: No significant difference was observed among groups in the calcium and inorganic phosphate concentrations in the fluid phase of the test plaque 30 minutes after the dentifrice use (P > 0.05), but F concentration was significantly higher for both F dentifrices (P < 0.05). Also, the dentifrices did not differ regarding the pH before or 5 minutes after the sugar challenge (P > 0.05). A higher mineral loss was observed for the non-F dentifrice group (P < 0.05), but no significant difference was observed between the F dentifrices containing CaGP or not (P > 0.05). Using this in situ model, the findings suggested that CaGP at the concentration tested did not enhance the inhibition of enamel demineralization promoted by F dentifrice.

Monitoring acid-demineralization of human dentine by electrochemical impedance spectroscopy (EIS).

OBJECTIVE: The purpose of this study was to monitor in situ acid-induced demineralization of dentine by using electrochemical impedance spectroscopy (EIS) and correlate the EIS findings with changes in the chemical composition and ultrastructure of dentine. METHOD: EIS was used to monitor the process of demineralization of dentine induced by an acid model. Scanning electron microscopy (SEM) was used to examine the ultrastructure, while energy dispersive X-ray (EDX) and X-ray diffraction (XRD) analysis were employed to investigate the changes in the chemical composition of surface dentine with demineralization. RESULTS: Two kinds of equivalent circuits that matched the histological structure of dentine were used to fit the EIS data. The parameters R(d), which is the resistance of the bulk of dentine layer and R(ct), which is the charge transfer associated with the penetration of electrolyte into the dentinal tubules, decreased with increasing duration of demineralization. SEM images showed that the smear layer on the dentine and peritubular dentine disappeared with demineralization. The EDX results showed that the content of calcium and phosphorus decreased consistently with the decreasing content of HAP as shown by the XRD results. The findings from this study suggested that the changes in R(d) and R(ct) determined by the EIS corresponded well with the variation in structure and composition of dentine. CONCLUSIONS: EIS could be employed to monitor structural and chemical compositional changes induced by acid-demineralization on dentine surface.

Effect of strontium in combination with fluoride on enamel remineralization in vitro.

Previous studies showed that strontium (Sr) as well as fluoride (F) can enhance enamel remineralization. The aim of this study was to evaluate the effects of Sr in combination with F on enamel remineralization in vitro. Sixty enamel specimens obtained from caries free human premolars were demineralised to produce caries-like lesions. Half of each lesion was covered with nail varnish as an untreated control. The specimens were then randomly divided into F and Sr+F treatment groups. The F group was exposed to remineralizing solutions (1.5mM CaCl(2), 0.9 mM KH(2)PO(4)) containing 1 ppm, 0.1 ppm or 0.05 ppm F. The Sr+F treatment group was exposed to the same solutions including 10 ppm Sr. After 2 weeks, lesion depth, mineral loss and percentage enamel remineralization were determined using transversal microradiography. There was a significant decrease in mineral loss in all groups (p<0.001). Lesion depth was significantly reduced for all groups (p<0.05) with the exception of group F. Remineralization was significantly affected by F concentration (p=0.000). The participation of Sr resulted in a significant enhancement of remineralization (p<0.001) with a synergistic effect of the Sr+F combination (p<0.01). It was concluded that while the remineralizing process was affected by the concentration of F, there was also an interaction between F and Sr when they were used in conjunction.

[Effect of Galla Chinesis on the demineralization of dental root tissue in pH cycling model].

OBJECTIVE: To evaluate the anti-demineralization efficacy of Galla Chinesis in pH cycling model for elucidating the anti-root caries mechanism. METHOD: Anti-demineralization efficacy evaluation of the natural medicine in the pH-cycling models was used . Sound human root blocks were pH-cycled through the treatment solution, acidic buffer and neutral buffer. The cycling times for demineralization study were 12 times, 2 times per day. The acidic buffers were retained for calcium analysis by atomic adsorption spectroscopy. The sections of blocks were analysed after pH-cycling by CLSM. Treatments were 4 g x L(-1). Galla Chinesis, 1 g x L(-1) NaF solution and distilled water. RESULT: Galla Chinesis was found to inhibit the demineralization in the pH cycling model. Although the effect was not as good as fluoride, there was no significant difference between the two groups. CONCLUSION: These data suggest that Galla Chinesis could modulate the mineralisation behaviour of root tissue in a defined chemical circumstance. These findings support the proposition that Galla Chinesis may be a promising anticaries natural medicine in the future.

Micromorphological and micronanalytical characterization of stagnating and progressing root caries lesions.

Root caries is the predominant disease of the tooth tissues in the elderly population and differs in progression and micromorphology from coronal dentin caries. Therefore, different clinical concepts are needed for the treatment of these progressing and arrested lesions. It was the aim of this study to investigate the three-dimensional structure and volume of stagnating and progressing root caries lesions and to determine the mineral composition of the different lesion zones to achieve a better understanding of the natural history of root caries lesion formation. Of 21 extracted human teeth of patients between 42 and 77 years of age with stagnating and progressing root caries lesions serial sections were cut and investigated with polarized light and scanning electron microscopy. From the polarized light micrographs 3D reconstructions were made to determine the volumes of the lesion zones and their relations expressed in a demineralization index (DI). With increasing size of the demineralizing zone the DI increased indicating an increase in the size of the translucent zone. The 3D reconstructions showed distinct differences between stagnating and progressing root caries lesions. In the hypermineralized translucent dentin not all dentin tubules were obliterated by intratubular dentin and within the translucent dentin scattered dead tracts were found. Electron dispersive X-ray (EDX) analysis showed that the intertubular dentin of the translucent dentin was also demineralized to a certain extent. The results indicate distinct morphological differences between progressing and stagnating root caries lesions which may have consequences for treatment strategies.

Ion uptake into demineralized dentine from glass ionomer cement following pretreatment with silver fluoride and potassium iodide.

BACKGROUND: Diamine silver fluoride (Ag(NH3)2F), referred to as AgF, has been shown to provide a pronounced antimicrobial action against caries. The clinical application of this material has been limited by the staining associated with both teeth and tooth coloured restorative materials. The application of potassium iodide (KI) after AgF eliminates stain formation. The purpose of this study was to determine if a prior application of silver fluoride and potassium iodine to demineralized dentine affected the uptake of strontium and fluoride from a glass ionomer cement restoration. METHOD: Three cavities were prepared in each of five recently extracted human third molars. The cavities were demineralized and treated as follows. In each tooth, one cavity was left as a control, one cavity was restored with glass ionomer cement and one cavity was treated with 1.8M AgF and a saturated KI solution and then restored with glass ionomer cement. The penetration of the various elements into demineralized dentine was measured by their relative percentage weights using electron probe microanalysis (EPMA). RESULTS: Fluoride uptake was significantly higher in the AgF and KI treated samples compared to the other two samples and significantly higher in the glass ionomer restored sample compared to the control. The application of AgF and KI did not significantly interfere with the transfer of strontium from glass ionomer cement into dentine. Silver and iodine deposits were present in the demineralized dentine treated with AgF and KI. CONCLUSIONS: The application of AgF and KI onto dentine prior to the placement of glass ionomer cement did not significantly affect the strontium uptake into the subjacent demineralized dentine and the fluoride levels in this zone were significantly increased.

Ca, Pi, and F in the fluid of biofilm formed under sucrose.

Calcium (Ca), inorganic phosphorus (P(i)), and fluoride (F) concentrations are low in the whole plaque biofilm formed under exposure to sucrose. It was hypothesized that this would be reflected in the biofilm fluid, where these low values should greatly influence the de/remineralization process. Dental biofilms were formed in situ over enamel blocks mounted in palatal appliances and exposed 8 times/day to distilled water, glucose+fructose, or sucrose solutions for 14 days. While Ca, P(i), and F concentrations in the whole biofilms were significantly lower in the glucose+fructose and sucrose groups, no effect on biofilm fluid was observed, even after a cariogenic challenge. An increase in whole biofilm mineral ions was observed 24 hrs after the carbohydrate treatments were suspended, but this effect was also not observed in the fluid. These results suggest that there is a homeostatic mechanism that maintains biofilm fluid mineral ion concentration, regardless of its total concentration in the whole biofilm.

Effect of iron on bovine enamel and on the composition of the dental biofilm formed "in situ".

OBJECTIVES: This study investigated in situ the effect of iron (Fe) on the reduction of demineralization of bovine enamel, as well as on the composition of dental biofilm. DESIGN AND METHODS: Twelve volunteers were included in this blind crossover study, which was conducted in two stages of 14 days each. For each stage, the volunteers received palatal appliances containing four blocks of bovine enamel (4 mm x4 mm x 2.5 mm). Six volunteers dripped a solution of 15 mmol L(-1) ferrous sulphate onto the fragments and the remaining six dripped deionized water (eight times per day). After five minutes, a fresh 20% (w/v) sucrose solution was dripped onto all enamel blocks. During the experimental period the volunteers brushed their teeth with non-fluoridated dentifrice. After each stage, the percentage of surface microhardness change (%SMHC) and area of mineral loss (DeltaZ) were determined on enamel and the dental biofilm formed on the blocks was collected and analysed for F, P, Ca, Fe and alkali-soluble carbohydrates. The concentrations of F, Ca and Fe in enamel were also analysed after acid biopsies. RESULTS: There was a statistically significant increase in the P and Fe concentrations in the biofilms treated with ferrous sulphate (p<0.05), which was not observed for F, Ca and alkali-soluble carbohydrates. The group treated with ferrous sulphate had significantly lower %SMHC and DeltaZ when compared to control (p<0.05). CONCLUSIONS: These results showed that ferrous sulphate reduced the demineralization of enamel blocks and altered the ionic composition of the dental biofilm formed in situ.

The remineralizing effect of an essential oil fluoride mouthrinse in an intraoral caries test.

BACKGROUND: The authors conducted a two-week clinical study to determine the remineralizing effect of an experimental mouthrinse containing both fluoride and essential oils in an intraoral caries test model. METHODS: The study used an observer-blinded, randomized, controlled, 3 x 3 crossover design. The authors enrolled in the study 153 subjects, each of whom had a mandibular removable partial denture. Two partially demineralized human enamel specimens were mounted on each subject's removable partial denture. Subjects used either a fluoride mouthrinse with essential oils (the test mouthrinse), a fluoride nonessential oils mouthrinse (the positive control) or an essential oil nonfluoride mouthrinse (the negative control) twice daily for 14 days. The researchers assessed specimens for mineral content change and fluoride uptake using surface microhardness, or SMH, testing and enamel fluoride analysis, respectively. RESULTS: Of the 153 subjects enrolled in the study, 125 subjects were evaluable at the study endpoint. The results after two weeks showed that percentage of SMH recovery was 42 percent in the test group, 36 percent in the positive control group and 16 percent in the negative control group. The fluoride uptake was 19 micrograms per square centimeter, 16 microg/cm2 and 3 microg/cm2 for the test mouthrinse, positive control and negative control groups, respectively. In terms of both percentage of SMH and fluoride uptake, the test mouthrinse and positive control mouthrinse were statistically higher than the negative control mouthrinse, and the test mouthrinse was "at least as good as" the positive control mouthrinse. CONCLUSIONS: This study provides evidence that an essential oil mouthrinse with 100 parts per million fluoride is effective in promoting enamel remineralization and fluoride uptake. CLINICAL IMPLICATIONS: The combination of fluoride and essential oils in a mouthrinse may provide anticaries efficacy, in addition to essential oils' previously established antigingivitis efficacy.

Nd:YAG laser ablation and acid resistance of enamel.

The acid resistance of Nd:YAG laser-ablated enamel surfaces was studied by evaluating crystal structure, mineral distribution, and fluorescence radiance and image in the present study. For comparison, 37% phosphoric acid etching was performed. The formation of beta-tricalcium phosphate (beta-TCP) was confirmed in the laser-ablated surface. The Ca/P ratio increased after ablation due to mineral re-distribution. In contrast, the Ca/P ratio decreased after acid etching due to mineral loss. The laser-ablated enamels showed a smaller increase of fluorescence radiances and less clear laser confocal scanning microscope images than those observed in the acid-etched enamels. The former suggests a minimized mineral loss. The Nd:YAG laser irradiation will enhance the acid resistance and retard the carious progression in enamel.

[The effect of Galla chinensis on the demineralization of enamel].

OBJECTIVE: To investigate the effect of Galla Chinensis with or without fluoride on demineralization of enamel. METHODS: Galla Chinensis with or without fluoride and acid buffer were used to treat the samples alternately and the treatment was repeated 8 times. Then the calcium of acid buffer was measured by American PE HTS 7000 plus and the specimens were observed with laser scanning confocal microscope. RESULTS: Compared with the calcium concentration of control group (deionized water or fluoride), the calcium concentration of all tested groups (except the 1 mg/ml group without fluoride) decreased significantly (P < 0.25). There was marked difference in total fluorescence (TF) and average fluorescence (AF) between the tested groups and control group (P < 0.05). As for the area of lesion, only groups without fluoride and the 4 mg/ml group containing fluoride decreased (P < 0.01). Confocal images showed that the zone of red area of tested groups was narrower than that of control group. CONCLUSION: Galla Chinensis can reinforce the resistance of enamel to acid, either with or without fluoride.