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An in vitro model for studying the efficacy of fluoride dentifrices in preventing root caries.

This investigation tested and validated an in vitro model for studying the effects of fluoride on human dentin mineral content and fluoride uptake. Four studies examined the effects of different concentrations of fluoride, established dose-response profiles with NaF and Na2PO3F, and tested commercial dentifrices. A 7- or 14-day cyclic treatment regimen involved four 1-min exposures of sound human dentin specimens to the treatment agents, a 4-hour acid challenge period, and 20 h in human saliva daily. Mineral content was analyzed by image analysis microradiography and fluoride assays were performed using a microdrill biopsy technique. Data from these studies established the ability of the in vitro model to provide reproducible results, to demonstrate significant dose-related differences in the effects of both NaF- and Na2PO3F-containing treatments on dentin fluoride uptake and demineralization, and to detect a fluoride-induced reduction in dentin caries, relative to a nonfluoride control, similar to results established in a clinical trial.

Biopsy↗

Inhibitive effect of a resin-modified glass ionomer cement on remote enamel artificial caries.

Glass ionomer cements (GICs) demonstrate the inhibition of caries lesions formed immediately adjacent to the restoration. This in vitro study was conducted to evaluate the distance at which a resin-modified GIC is able to exert its cariostatic effect on artificial enamel lesions ('remote effect'). Resin-modified GIC or bis-GMA resin was applied on the cervical third of the labial surface of 10 paired halves of bovine incisors. Specimens were separately immersed for 3 weeks in lactic acid gel which was changed every other day to reduce fluoride accumulation. Artificial lesions were examined by the cross-sectional microhardness (MHN) method. Volume percent mineral, mineral loss (delta Z value) and change in mineral content (delta M) were computed for each MHN profile, performed at distances of 0.2, 0.5, 1.0, 2.0, 4.0 and 7.0 mm from the edge of the materials. delta Z values of the resin-modified GIC group were significantly lower than those of the bis-GMA control group at all remote sites (t test, p < 0.05). The delta M caused by resin-modified GIC was more pronounced within 1.0 mm from the material which suggested that the demineralization inhibition can be divided into the near effect (< 1.0 mm). In this in vitro study, resin-modified GIC provided caries resistance in bovine enamel located at a considerable distance from the margin of the material.

Animals↗

In vitro evaluation of the cariostatic action of esthetic restorative materials in bovine teeth under severe cariogenic challenge.

Considering that caries around restorations is a serious problem in dentistry, and some restorative materials with fluoride may be important in inhibiting these lesions, this research is aimed at performing an in vitro evaluation of the cariostatic action of some esthetic restorative materials. Standardized cavities were prepared in the center of either intact blocks of bovine enamel or with bovine teeth containing early artificial carious lesions. The specimens were restored with a high viscosity glass ionomer cement (Molar Ketac), a resin-modified glass ionomer cement (Vitremer), a polyacid-modified resin composite (Dyract AP) and a conventional resin composite (Z-250). In addition to the restored specimens, four corresponding control groups were evaluated. All groups, except for two control groups, were subjected to a demineralization/remineralization cycling model for 14 days, simulating a situation of severe cariogenic challenge. The blocks were then longitudinally sectioned through the restorations. Mineral loss was evaluated in these specimens using the Knoop microhardness profiles in longitudinal sections at three different distances of the cavities and at eight distinct depths in relation to the external enamel surface. Statistical analysis of the results showed significant differences (p < 0.05) among the groups, although none of the study materials completely inhibited creation of the lesions. Vitremer demonstrated the best cariostatic action in intact bovine enamel. Ketac Molar, in intact or demineralized enamel, and Vitremer, in demineralized enamel, presented intermediate cariostatic potential. Z-250 and Dyract AP did not demonstrate any cariostatic effect. The data suggests that glass ionomer cements demonstrated better cariostatic action compared to the other restorative materials.

Analysis of Variance↗

[The effect of the exploratory probe on demineralized enamel].

This study aims to demonstrate the effect of dental explorer on demineralized enamel. Bovine teeth were divided perpedinculary towards their long axis. Each section were provided with 2 grooves, parallels to the teeth's axis. These sections were exposed to an acid solution for 7 days. After this period just one of the grooves of each section was probed ahd then, the demineralization was continued for one more wee. Then we could compare the lesions depths of demineralization of the probed grooves and the non probed grooves. The probed grooves presented an increased lost of enamel structure, more than the non probed grooves. It was concluded that the careless use of dental explorer on diagnosis of fissures caries cause mechanical damage to enamel already softened as a result of a carious attack.

Animals↗

Effects of low fluoride concentrations on formation of caries-like lesions in human enamel in a sequential-transfer bacterial system.

Human enamel surfaces were exposed to sequential batch cultures of Streptococcus mutans NCTC 10832 in a sucrose-rich medium containing 0-5 mg/l added fluoride (F). In 10-day experiments, subsurface lesion formation was partly inhibited by 1 mg/l F and completely by 2 and 5 mg/l F, but small lesions formed in 2 mg/l F in 21-day experiments. Analysis of the spent media, together with analogous, bacteria-free experiments, suggested that lesion inhibition involved two main effects. First, inhibition of bacterial acid production reduced the pH fall, resulting in reduced undersaturation with respect to hydroxyapatite and consequently reduced rate of demineralization. Secondly, interaction of F with enamel mineral resulted in a small increase in reprecipitation during periods of supersaturation and a much larger reduction in demineralization during periods of undersaturation. It is concluded that, at low F concentrations, inhibition of bacterial acid production is a major factor in lesion inhibition, which may contribute significantly to caries prevention in vivo where plaque fluid F levels are raised by frequent topical applications.

Apatites↗

Caries inhibition around composite restorations by pulsed carbon dioxide laser application.

This in vitro study aimed to evaluate whether laser irradiation of cavity margins reduces enamel demineralization around composite restoration. Enamel cavities were prepared in 33 human enamel slabs, which were randomly divided into three groups. One group was kept as a control, and the cavosurface margin of the cavities of the other groups were irradiated, using a CO(2) laser (lambda = 10.6 microm), at 8 J.cm(-2) or 16 J.cm(-2). The cavities were restored with a resin-based composite, according to the manufacturer's specifications. Before restoration, scanning electron microscopy was performed on one specimen of each group. The remaining slabs were submitted to thermal and pH-cycling models. Enamel mineral loss, at 50 and 100 microm from the restoration margin, was assessed by cross-sectional microhardness analyses. Fusion and melting were observed in the irradiated groups. Mineral loss at 50 microm from the restoration margin was significantly inhibited in the irradiated groups compared to the control group, but at 100 microm from the restoration margin, mineral loss at only the highest laser energy density differed statistically from the control group. The difference between the irradiated groups was not statistically significant at either 50 or 100 microm from the restoration margin. In conclusion, irradiation of the cavosurface margin of cavities, using a pulsed CO(2) laser, is able to inhibit enamel demineralization around composite restorations, and an energy density of 16 J.cm(-2) is efficient, even at 100 microm from the cavity margin.

Bisphenol A-Glycidyl Methacrylate↗

Mineral distribution and dimensional changes in human dentin during demineralization.

Many bonding agents require the dentin surface to be acid-etched prior to being bonded. Understanding the stability and morphology of the etched dentin surface is important for improving bond strength and reliability in these systems. In this study, the atomic force microscope was used to quantify dimensional changes that occur to fully hydrated dentin during demineralization with a pH 4.0 lactic acid gel. A high-resolution microtomography instrument, the x-ray tomographic microscope, was also used to quantify the mineral density distribution in the dentin as a function of etching time. The intertubular dentin surface shrank by less than 0.5 microns during etching, while the peritubular dentin receded at an initially rapid linear rate. The dentin surface retained its initial morphology, although it was more porous with the removal of the peritubular dentin. Beneath the etched surface, there were three major zones characterized by mineral density differences. The first zone was a fully demineralized collagen layer, subjacent to which was a partially demineralized zone of roughly constant mineral density. Immediately following the partially mineralized layer was normal dentin. The presence of the partially mineralized layer could be explained in terms of different transport rates in the peritubular and intertubular dentin.

Acid Etching, Dental↗

Fluoride-releasing capacity and cariostatic effect provided by sealants.

This study evaluated the effect of sealants on enamel demineralization, focusing on physical protection of the sealed enamel and fluoride protection of the adjacent unsealed enamel. Occlusal fissures with areas measuring 12 mm2 were delimited in 48 extracted molars, randomly divided into 4 groups (n =12): 1) no sealing; 2) sealing with a resin-modified glass-ionomer (Vitremer, 3M ESPE); 3) sealing with a fluoride-releasing composite sealant (Clinpro Sealant, 3M ESPE); and 4) sealing with a non-fluoridated composite sealant (Concise, 3M ESPE). A 4-mm2 window was outlined on the buccal enamel for analysis of fluoride uptake. Following treatment, groups 2, 3 and 4 were subjected to 5-days of pH-cycling, while group 1 was kept in a moist environment at 37 degrees C. Fluoride uptake was assessed by dental biopsy, and the amount of fluoride released to the cycling solutions was determined by ion analysis. Enamel demineralization around the sealants was evaluated by cross-sectional micro-hardness analysis. Group 2 showed higher levels of fluoride release (P < 0.01) and uptake by enamel (P < 0.05), and lower levels of demineralization (P < 0.05) than groups 3 and 4. Group 3 exhibited reduced demineralization on unsealed enamel and provided fluoride uptake in a distant enamel area, while group 4 did not.

Analysis of Variance↗

Inhibition of dentin demineralization by fluoride in vitro.

Compared with the knowledge accumulated on enamel-fluoride interactions, relatively little data is available regarding fluoride effects on dentin. This applies to both laboratory and clinical studies into the efficacy of fluoride schemes for the prevention of root surface caries. This study aimed to determine the effects of fluoride and pH on the demineralization of dentin, such as to provide information necessary to develop preventive programmes. Bovine dentin blocks were subjected to undersaturated calcium- and phosphate-containing solutions in the pH range 4.0-6.0 with fluoride added at concentrations between 0.5 and 10 ppm. Non-fluoride solutions served as controls. Mineral loss was assessed chemically and by transversal microradiography. Comparisons were made with similar studies on enamel demineralization. The results showed that demineralization of dentin depends on both pH and fluoride concentration in the demineralizing solution. Inhibition of demineralization that could be relevant from a clinical point of view was found at fluoride values 5-10 times the corresponding values for enamel. Also rapid depletion of fluoride from the solutions was observed, indicating the high uptake capacity of dentin for fluoride. Lesion depth depended on pH of the solution while the fluoride levels were associated with the surface layer, both in mineral content and depth. For dentin we propose a demineralization mechanism where acid penetrates rapidly into the tissue, presumably through the tubules, after which the released calcium and phosphate is partly trapped by the inward diffusing fluoride. This leads to the formation of a surface layer, which may even be hypermineralized compared to sound dentin.

Animals↗

Effect of frequency of sucrose exposure on dental biofilm composition and enamel demineralization in the presence of fluoride.

It has been suggested that enamel would resist higher frequencies of sucrose exposure if fluoride from water or dentifrice is being used. However, the effect of increasing frequencies of sugar on dental biofilm composition is not well known. Ten volunteers living in a fluoridated area wore palatal appliances bearing human enamel slabs during 14 days. The slabs were exposed to 20% sucrose solution 0 (control), 2, 4, 6, 8 or 10 times/day and the volunteers used fluoride dentifrice 3 times/day. Enamel demineralization was significantly greater than control for sucrose frequencies higher than 6 times/day. However, biofilm mass, total microbiota, total streptococci, lactobacilli counts and insoluble extracellular polysaccharide concentration increased, while Ca, P(i) and F concentration in whole biofilm decreased significantly, with frequencies of sucrose exposure lower than 6 times/day. The findings confirm that fluoride can reduce enamel demineralization if sucrose consumption is not higher than 6 times/day, but changes in the biochemical and microbiological composition of the biofilm are observed with lower frequencies of sucrose use.

Adolescent↗

An improved intra-oral enamel demineralization test model for the study of dental caries.

The intra-oral enamel demineralization test (IEDT) was introduced by Brudevold et al. (1984). This caries model involves human subjects wearing palatal appliances each holding eight bovine enamel blocks covered by a bacterial cell layer prepared by the harvesting of cultures of Streptococcus mutants (test plaque). The original model used the iodide permeability test for assessment of the extent of demineralization of bovine enamel blocks resulting from acid production by the test plaque after dietary substrate challenge. The IEDT model has been expanded and improved by us in the following ways: (1) Based on encouraging findings from an in vitro study (Zero et al., 1990), the surface microhardness test has been adopted to measure the extent of demineralization occurring at three sites on the enamel blocks corresponding to an area over which the effective plaque thickness is 0.5, 1.5, and 2.5 mm; (2) intra-oral pH of the test plaque is measured by means of a Beetrode miniature pH electrode at baseline, then at five, 10, 15, 30, and 45 min after the start of a test; (3) plaque samples are collected at the end of a test and analyzed for organic acid content by means of HPLC; (4) the bacterial test challenge has been expanded to include different cariogenic bacteria which are grown under various growth conditions. The improved model has the capability of studying fundamental aspects of the caries process, namely, the relationships among dietary substrate challenge, plaque pH change, plaque organic acid profiles, microbial virulence properties, and enamel demineralization. Furthermore, the model has the potential for use in more applied research on caries-preventive agents such as fluoride.

Adult↗

Inhibition of bovine dentin demineralization by a glutardialdehyde pretreatment: an in vitro caries study.

Intact bovine dentin specimens were demineralized in 25 mmol acetic acid buffers (pH 5), with and without a 5-min pretreatment with an acidic 2% glutardialdehyde (GDA) solution. The results demonstrate that GDA inhibits dentin demineralization and that the inhibition depends strongly on the initial calcium and phosphate concentration in the demineralization solution. The observed inhibition increases from about 5% without calcium and phosphate to 38% with a calcium and phosphate concentration initially of 6.4 mmol and 4.1 mmol, respectively. The inhibition can be explained either by a reduction of calcium and phosphate diffusion out of the lesion or by the retention of mineral inhibitors in the dentin.

Animals↗

Fluoride release and caries inhibition associated with a resin-modified glass-ionomer cement at varying fluoride loading doses.

PURPOSE: To measure, over time, the release of fluoride from a resin-modified glass-ionomer cement, loaded with different levels of fluoride, then to evaluate the adjacent dentin demineralization inhibition relative to these fluoride levels. MATERIALS AND METHODS: (Phase I) 25 standardized discs were fabricated from a non-fluoridated resin-based composite (control), resin-modified glass-ionomer cement and resin-modified glass-ionomer cement loaded with sodium fluoride at 1%, 2% and 3% by weight fluoride. Fluoride release was evaluated over 30 days. (Phase II) 50 restorations, from the materials listed in Phase I, were placed as Class V restorations in teeth, the teeth were acid-challenged, then dentin margins adjacent to restoration margins were evaluated for demineralization. RESULTS: As sodium fluoride additions increased, fluoride release increased. Evaluation of demineralization indicated the resin-modified glass-ionomer cement inhibited adjacent demineralization in a direct relationship with sodium fluoride concentration where 3% fluoride exhibited significantly less adjacent demineralization than all other groups; 2% and 1% fluoride exhibited significantly less adjacent demineralization than the non-loaded resin-modified glass-ionomer cement and non-fluoridated resin-based composite control (P< 0.05).

Acid Etching, Dental↗

High resolution electron microscopy of enamel crystallites demineralized by initial dental caries.

Acids produced by various oral bacteria cause mineral loss and crystallite dissolution during the development of enamel caries. In order to demonstrate this phenomenon, the initial disappearance of lattice fringes and the formation of a central perforation in crystallites were examined by high resolution electron microscopy (HREM) in initial enamel caries without macroscopic tissue evidence of destruction. Ultrathin sections were also examined by selected area electron diffraction to reveal the mineral phase of the surface layer in carious enamel. A marked variation in the dissolution pattern was disclosed in the initial carious lesions. HREM revealed that disappearance of the lattice fringes from the lateral portion of the crystallites was predominant in the superficial layer covering the lesion, while central perforation of crystallites mainly occurred in the subsurface prismatic region. The beginning of the central dissolution occurred at the dislocated area where lattice striations appeared to be disordered. Selected area electron diffraction of the gradually demineralized enamel revealed a pattern consistent with hydroxyapatite (OH-AP) or fluorapatite (F-AP) mineral. These findings suggest that the susceptibility to caries of enamel crystallites is spatially and temporally different during the progression of the caries. Furthermore, the formation of central perforations and the consequent easy intracrystalline diffusion of acids might induce rapid crystallite dissolution.

Adult↗

Ability of restorative and fluoride releasing materials to prevent marginal dentine demineralization.

The study aimed to define the in vitro secondary caries inhibiting potential of restorative materials currently used in dental practice. Class V restorations were prepared in extracted human third molars and immersed in a demineralizing solution (lactic acid, pH 4.5) at 37 degrees C for 2 days to simulate secondary caries formation. The bonding and the restorative systems tested in the study were: Scotchbond 1+Z 250 (Group A), Scotchbond 1+F 2000 (Group B), ABF+APX (Group C), ABF+F2000 (Group D). Perimarginal dentine, immediately close to the margin of the restoration, and exposed dentine, at approximately 0.5 mm from the margins of the restoration, after exposure to the acid solution, were investigated; protected dentine, at approximately 4 mm from the margin in a varnish-covered area, was analysed as control. Polarized light microscopy and contact transverse microradiography (TMR) were employed. The output parameters were lesion shape and size (depth in microm) of the exposed dentine, dentine mineral volume%, and integrated mineral loss (Delta Z, in %volmicrom) of the lesions. Compomers (Groups B and D) showed a thinner demineralization of the outer lesions, a less demineralization along the perimarginal dentine (inner lesion) and more caries inhibition zones or CIZs (Delta Z positive values) compared to composites (Groups A and C). In conclusion, Groups B and D materials seemed to partially counteract the marginal demineralization induced by an acid solution and favourably influence the formation of CIZs along the restorations. On the contrary, composites did not show a protective effect, probably due to an insufficient marginal seal and the lack of fluoride release.

Absorptiometry, Photon↗

Submandibular gland aplasia and progressive dental caries: a case report.

When severe caries occurs in mandibular permanent incisor teeth, the clinician should consider the possibility of associated submandibular gland aplasia or salivary hypofunction. Early diagnosis of submandibular gland disease is essential, as operative problems involving restoration of mandibular incisor teeth are considerable. Furthermore, progressive severe dental caries can present a dilemma for the clinician in affected individuals, despite intensive preventive and restorative therapy. A case report describing severe progressive dental caries and enamel demineralization of the permanent mandibular incisor teeth in a young girl is presented. Further investigation revealed absence of functional bilateral submandibular salivary glands contributing to the rapid breakdown of the teeth despite intensive preventive measures.

Adolescent↗

Effect of calcium glycerophosphate on demineralization in an in vitro biofilm model.

The aim was to investigate the anti-caries properties of calcium glycerophosphate (CaGP) using an in vitro bacterial flow cell model. Four flow cells, inoculated from a chemostat containing a seven-organism bacterial consortium, were pulsed with sucrose twice daily, to provide an acidic challenge and pH-cycling conditions. Blocks of enamel and dentine were mounted in each flow cell. In a study on the effect of CaGP concentration, CaGP was pulsed into three of the flow cells, at the same time as the sucrose, to give concentrations of 0.10, 0.25 and 0.50%. Water was pulsed into the fourth flow cell with the sucrose. Microradiography revealed a significant dose response of decreasing demineralization as CaGP concentration increased. Reductions at 0.25 and 0.5% were significant when compared to the control. A second study investigated the effect of timing of CaGP pulsing, relative to sucrose, on enamel and dentine demineralization. CaGP (flow cell concentration 0.2%), was pulsed 1 h before, during or 1 h after the sucrose pulse; a water control was employed. In enamel, pulsing CaGP before the sucrose reduced demineralization significantly compared to concurrent pulsing, which in turn gave a significant reduction compared to pulsing after sucrose, which did not reduce demineralization significantly compared to the water control. In dentine, CaGP reduced demineralization significantly only when pulsed before the sucrose. The findings suggest that in vivo, the anti-caries potential of CaGP may be greater if it is applied before a cariogenic challenge.

Actinomyces↗

Continuing etching of an all-in-one adhesive in wet dentin tubules.

Self-etch adhesives that etch and prime simultaneously are becoming more acidic. We hypothesized that the degree of acidic monomer conversion at the interface and within the tubules was high enough that the acidic reaction would be very self-limiting. Dentin surfaces prepared from extracted, unerupted human third molars were treated with Prompt L-Pop (3M ESPE). The prepared teeth were stored in normal saline, and specimens retrieved at intervals < or = 4 wks were randomly selected for light, scanning electron microscopic and micro-Raman spectroscopic analysis. Morphologic and spectroscopic analyses indicated dentin demineralization and adhesive penetration throughout the demineralized layer and tubules. Increased dentin demineralization and loss of adhesive integrity were noted after aqueous storage. The degree of monomer conversion at the interface was consistently greater than conversion within the tubules. Fluid within the tubules may inhibit monomer conversion. The acidic characteristics of this adhesive may be retained and, thus, continue to affect/demineralize the surrounding dentin.

Acid Etching, Dental↗