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Mineral density and fluoride content of in vitro remineralized lesions.

This in vitro investigation studied the remineralization of experimental caries lesions in bovine enamel by use of three methods: (1) surface microhardness, (2) microradiography, and (3) abrasion biopsy for mineral density and fluoride content. The lesions were produced by a two-day exposure to 0.01 mol/L lactic acid/sodium hydroxide buffer partially saturated with 3.0 mmol/L Ca, 1.8 mmol/L P, in 1% CMC, at pH 4.0 and 37 degrees C. The lesions were exposed to a remineralizing solution containing 3.0 mmol/L Ca, 1.8 mmol/L P, and 3 ppm F in 1% CMC at pH 7.0 and 37 degrees C for two, six, and ten days, with solution changes every two days. The data derived from the three methods are presented in sequence from the baseline and at days two, six, and ten of the remineralizing treatment. Microhardness measurements showed hardness recoveries of 35.9, 78.9, and 87.5%, respectively. Microradiography suggested complete recovery with the ten-day remineralization. Abrasion biopsy of successive 10-micron layers to a depth of 100 micron indicated 15.2, 39.8, and 68.8% mineral density recoveries, with fluoride content of the surface layer increasing from a baseline of 300 ppm to 4600, 9000, and 9800 ppm F for the 2, 6, 10 days of remineralization, respectively. Subsequent acid-etching of thin sections from the ten-day-remineralized specimens showed that the fluoride-enriched remineralized area was more resistant to acid dissolution than was the underlying normal enamel.

Animals↗

Remineralization of enamel subsurface lesions by casein phosphopeptide-stabilized calcium phosphate solutions.

Casein phosphopeptides (CPP) stabilize amorphous calcium phosphate (ACP), localize ACP in dental plaque, and are anticariogenic in animal and in situ human caries model. In this vitro study, CPP-stabilized calcium phosphate solutions were shown to remineralize subsurface lesions in human third-molar enamel. Solutions were used to examine the effect of CPP-calcium phosphate concentration on remineralization. Other solutions were used to examine the effect of increasing pH, which decreased the concentrations of free calcium and phosphate ions and increased the level of CPP-bound ACP. Although most of the remineralizing solutions were supersaturated with respect to the amorphous and crystalline calcium phosphate phases, the solutions were stabilized by the CPP such that spontaneous precipitation of calcium phosphate did not occur. After a ten-day remineralization period, enamel lesions were sectioned, subjected to microradiography, and the mineral content determined by microdensitometry. All solutions deposited mineral into the bodies of the lesions, with the 1.0% CPP-calcium phosphate (pH 7.0) solution replacing 63.9 +/- 20.1% of mineral lost at an averaged rate of 3.9 +/- 0.8 x 10(-8) mol hydroxyapatite/m2/s. The remineralizing capacity was greater for the solutions with the higher levels of CPP-stabilized free calcium and phosphate ions. Remineralization was not significantly correlated with either the CPP-bound ACP of the degrees of saturation for hydroxyapatite, octacalcium phosphate, or ACP. However, remineralization was significantly correlated with the degree of saturation for dicalcium phosphate dihydrate (CaHPO4.2H2O), but his was attributed to the significant correlation of remineralization with the activity gradients from the solution into the lesion of some calcium phosphate ions and ion pairs, in particular the neutral ion pair CaHPO4(0). The CPP, by stabilizing calcium phosphate in solution, maintain high-concentration gradients of calcium and phosphate ions and ion pairs into the subsurface lesion and thus effect high rates of enamel remineralization.

Calcium↗

[Development and laboratory analysis of new remineralizing means].

In laboratory research there were studied different combinations of remineralizing agents allowing production of substances close by chemical composition to hydroxyapatite of hard dental tissues. The most acceptable for practical use in case of demineralizing foci of hard dental tissues treatment can be only 2 calcium orthophosphates that may be synthesized under physiological conditions from solutions of calcium nitrate and twice-substituted ammonium phosphate, in particular, dicalcium phosphate dihydrate (structure of brushite, CaHP0(4) x 2H(2)O) and the so called precipitated hydroxyapatite (Ca(10-x) (HPO(4))(x)(PO(4))(6-x)(OH)(2-x)). The data are the bases for development of remineralizing agents and their further experimental and clinical studies.

Biocompatible Materials↗

Remineralization effects of xylitol on demineralized enamel.

We morphologically determined the effects of xylitol on the remineralization of artificially demineralized enamel. The samples were demineralized and then immersed in a remineralizing solution with or without 20% xylitol at 37 degrees C for 2 weeks. Samples were observed using contact microradiography, a multipurpose image processor (MIP) and a high-resolution electron microscope (HRTEM). Contact microradiography indicated that remineralization occurred in the surface and deep layers of samples immersed in a non-xylitol solution. Samples immersed in a xylitol solution demonstrated less mineralization in the outer 10 microm of the outermost surface layers, but more mineralization in the middle and deep layers, than was observed in the non-xylitol samples. The MIP evaluation indicated that remineralization was more prominent in layers at depths of 50-60 microm in the xylitol samples than in the non-xylitol samples. Observation of the xylitol samples by HRTEM revealed crystals of various sizes and irregular shapes with unclear crystal angles in the outermost surface layers. In the middle layers, they had thickened and angles of crystals are clear. These results indicate that xylitol can induce remineralization of deeper layers of demineralized enamel by facilitating Ca2+ movement and accessibility.

Crystallization↗

pH-cycling of enamel and dentin lesions in the presence of low concentrations of fluoride.

F-dentifrice usage causes slightly elevated fluoride levels in saliva. Therefore, the effects of permanent low fluoride concentrations versus daily dentifrice treatments were studied on enamel and dentin lesions in a pH-cycling model of alternating demineralization and remineralization. Groups received 1) no fluoride treatment, 2) 3 mu M (0.06 ppm) F continuously present during re- and demineralization or 3) daily 5-min F-dentifrice treatments. Solutions were analyzed for changes in calcium and fluoride. Cumulative results (10 d) showed that for the non-fluoride group the dentin lesions increased, while for enamel lesions mineral uptake and loss were balanced. Addition of 3 mu M F caused small, non-significant, enhancement of remineralization (1-7%), while demineralization was significantly inhibited for both tissues (9-23%). The daily dentifrice treatments resulted in a balance between mineral uptake and loss of dentin, due to inhibited demineralization (-33%) and enhanced remineralization (+79%). For enamel, the F-dentifrice treatments resulted in 43% reduction of demineralization, with no significant effect on remineralization. Fluoride loss from the 3 mu M F cycling solutions was significant (up to 50%) and constant during the experimental period. Microradiographic analysis showed remineralization at the lesion front in enamel. In dentin, the lesion depth was increased in all groups, with concomitant mineral deposition in the surface region of the dentifrice group. Results indicate that slightly elevated fluoride levels may be considerably less effective in inhibiting lesion progression in dentin than in enamel, and suggest mineral uptake and loss to occur at similar depths for enamel lesions, while demineralization and remineralization occur at different depths in dentin.

Animals↗

Remineralization of softened bovine enamel following treatment of overlying plaque with a mineral-enriching solution.

Pre-softened, gauze-covered bovine enamel blocks were worn in the buccal sulcus of five subjects for seven days. Artificial plaque enmeshed in the gauze was treated four times per day for four days with an enzyme-dependent mineralizing solution, resulting in 20-, 10-, and 200-fold increases in Ca, P, and F, respectively. Enamel beneath this mineral-enriched plaque recovered 37% of the hardness lost from pre-softening, while control enamel beneath untreated plaque recovered only 14%. Test enamel contained from five to 13 times as much F as did control enamel in the outer four layers sampled. Even though direct use of the mineralizing solution without the interposition of plaque caused a hardness recovery and F uptake similar to those in test enamel in vivo, a direct solution effect on enamel is not thought to explain the in vivo effects. Plaque treated with the urea-containing solution rapidly reaches a pH greater than 8, when a precipitate develops, leaving much-reduced ion concentrations in solution. Direct exposure of softened enamel to such a supernatant resulted in reduced hardness recovery and F uptake. It is concluded that the in vivo enamel remineralization was due mainly to the presence of a mineral phase in the overlying plaque. This mineral could have promoted remineralization by creating mildly supersaturated conditions during normal plaque pH cycles.

Adult↗

Enamel remineralization: how to explain it to patients.

The term remineralization of initial enamel caries is frequently used, but mainly at research conferences and in lecture rooms. Clinicians avoid discussing remineralization and white-spot enamel lesion formation with patients because details of the processes are complex. Patients, therefore, incorrectly assume that cavitation occurs right at the onset of caries and that a restoration must be placed to halt further progression of the lesion. Although laboratory and clinical studies have shown that initial white-spot enamel lesions can remineralize, patients have little or no access to this information. The schematic diagrams in this paper explain the diffusion of organic acids from plaque between enamel crystals, the partial loss of mineral from enamel crystals, and subsequent demineralization and remineralization that produces the white-spot lesion. Further, these diagrams may help clinicians explain remineralization as simply as possible to patients who present with white-spot enamel lesions.

Dental Caries↗

Quantitative measurements of remineralization of incipient caries.

White demineralized areas after the removal of orthodontic appliances remain a problem for clinicians and patients. The main objective of this study is to assess clinical photography as a method for the evaluation of caries and white spots and the in vivo study of remineralization of carious lesions. The secondary objective was to compare the effect of different levels of both experimental and commercial fluoride solutions on the remineralization of enamel carious lesions. White spot lesions were created with an acid solution on extracted permanent human teeth, and the lesions were remineralized in remineralizing solutions with or without fluoride ions. The changes in the enamel surface during the demineralization and the remineralization processes were recorded with a 35 mm clinical camera. Photographs were taken of the experimentally created white spots, the samples were sectioned for microradiography testing, and the actual mineral contents of the white spot lesions were calculated from the microradiographs. The enamel lesions were further analyzed by powder x-ray diffraction to confirm whether fluoride was incorporated into the lesions during the remineralization period. From this experiment it can be concluded: (1) clinical photography as currently practiced is not an adequate method of monitoring the remineralization of white spots with large lesion depths and (2) the experimental solution that contains 50 ppm fluoride had a higher efficacy for remineralization than the control solution or the commercial mouth rinse, which contained 225 ppm fluoride.

Crystallography, X-Ray↗

Experimental intra-oral caries models in fluoride research.

The use of experimental intra-oral caries models has increased in fluoride research. This paper focuses on the pre-clinical intra-oral models, the in situ and in vivo models, the various types, their benefits and disadvantages. Both preparation and sterilization of the hard tissue substrates can affect the substrates and therefore the results. Care needs to be taken that dentine samples are not exposed to drying and consequently shrinking during preparation and evaluation. Sterilization by gamma-radiation is at present the least tissue-damaging method. The most realistic experimental model is the in vivo model, followed by the in situ model using specimens with natural surfaces. The most accurate and direct evaluation technique for demineralization and remineralization studies is quantitative transversal microradiography, whereas confocal laser scanning microscopy (CLSM) is the most sensitive qualitative evaluation technique. Other evaluation techniques discussed are microhardness testing and the iodine permeability test. In light of the present skewed caries situation in western countries we suggest that fluoride research focuses on experimental caries models that can mimic severe cariogenic challenge. Testing of fluoride combinations and dosages that can prevent lesion development rather than promote remineralization would then be a practical consequence.

Animals↗

Use of variable remineralization periods to improve the abrasion resistance of previously eroded enamel.

The objective of the study was to evaluate the period of remineralization needed to re-establish the resistance of eroded enamel against brushing abrasion. Enamel specimens were prepared from 84 polished bovine incisors. The samples were evenly distributed among 7 groups (A-G) and submitted to ten alternating de- and remineralization cycles which included abrasion by toothbrushing. Demineralization was accomplished by immersing the specimens in the erosive soft drink Sprite Light((R)) for 1 min. Remineralization was performed by storing the samples in artificial saliva for either 0 min (A), 10 min (B), 60 min (C) or 240 min (D). After each remineralization, the specimens were brushed in an automatic brushing machine. Group E (erosion and 240 min remineralization, but no brushing) group F (erosion, but no remineralization and no brushing), and group G (brushing, but no erosion and no remineralization) served as controls. After performing the cycles, loss of enamel was determined by profilometry. The following values (mean +/- SD) were obtained and statistically analyzed (p<0.05): group A (5.16+/-1.26 microm), B (2.47+/-0.68 microm), C (1.72+/-0.75 microm), D (1.11+/-0.42 microm), E (0.81+/-0. 23 microm), F (1.04+/-0.31 microm), G (0.22+/-0.15 microm). Only the differences between groups D, E, and F were statistically not significant. Under the chosen in vitro conditions, it is concluded that abrasion resistance of eroded enamel continuously increases with remineralization time. However, even after a period of 1 h of remineralization, abrasion of previously eroded enamel is increased

Analysis of Variance↗

Remineralization of bovine dentine in vitro. The influence of the F content in solution on mineral distribution.

In this paper remineralization of bovine dentine is reported. After demineralization of the dentine in an acidic gel system creating lesions of about 180 microns depth, the tissue was remineralized in a 1.5 mM Ca and 0.9 mM phosphate containing solution at pH 7 and 37 degrees C for 8 or 21 days. The F content in the remineralization solution was 0.02, 2, or 10 ppm as NaF. Samples were analyzed by means of microradiography and scanning electron microscopy. The results show that remineralization (without F added in solution) causes a decrease in mineral loss and in lesion depth. With 2 or 10 ppm in solution, however, a substantial mineral accumulation in the lesion, but particularly on the original dentine surface, was observed. During 3 weeks of remineralization, mineral accumulations of 67 and 70 vol% of mineral for 2 and 10 ppm F in solution, respectively, were found near the original outer surface, with the sound dentine mineral value being 48 vol%. The combined microradiography and scanning electron microscopy data show that the mineral accumulated is for a major part deposited on the dentine tissue and partly inside. The latter deposition occurred both inside the tubules as well as in the intertubular areas. Inside the tubules dense precipitates were observed. Because several differences exist between bovine and human dentine, an extrapolation to the human in vivo situation is speculative. The results indicate that the presence of fluoride in the parts per million range is important for dentine remineralization efficacy and that the outer surface area in dentine can be 'overremineralized'.

Animals↗

Development of an intra-oral single-section remineralization model.

An intra-oral model system has been developed which uses a single-section technique for before-and-after measurements on the same tissue. This model allows for placement of sections of enamel at both buccal and approximal sites. These sections may be sound or possess white-spot lesions. This allows for the evaluation of lesion initiation, lesion progression, and/or lesion remineralization. The sections are appropriate for measurement by polarized light microscopy, and, although not included in this study, they are also suitable for microradiographic evaluation. With ten human volunteers, the model system was evaluated by means of three rinse regimens in a cross-over design. The distilled water rinse showed the whole range of possible changes (e.g., demineralization, no change, and remineralization). More consistent remineralization was observed when a fluoride rinse or a remineralizing fluid was used. This model system should prove to be quite useful in evaluation of demineralization and remineralization phenomena in the oral environment.

Adult↗

Effect of Various Types of Remineralizing Agents on the Remineralization of Mild and Moderate Defects Caused by Molar-Incisor Hypomineralization in Mexican Schoolchildren: A 12-month Follow-up Randomized Clinical Trial.

PURPOSE: Using Laser-Induced Fluorescence (LIF), this study aimed to evaluate the effect of three remineralizing agents on permanent molars and incisors with both mild and moderate Molar-Incisor Hypomineralization (MIH) in 8-to-12-year-old Mexican schoolchildren. MATERIALS AND METHODS: In this randomized clinical trial, 104 children were selected to participate and then divided randomly into four groups: Group I: control, Oral-B kids (toothpaste); Group II: Fluor Protector (varnish); Group III: Clinpro White Varnish; and Group IV: MI Paste (CCP-ACP). Both varnishes were applied once every four weeks for twelve months for each group. Remineralization was evaluated by LIF at intervals of 1, 3, 6, 9, and 12 months. Paired-sample t-tests and repeated-measures ANOVA were performed, while the Greenhouse-Geisser correction was used to evaluate sphericity. RESULTS: The three test agents used increased the level of remineralization observed at the site of mild and moderate MIH defects over the course of twelve months (p0.001). A similar decrease in average LIF values was observed across the three treatment groups. CONCLUSIONS: The use of both fluoride varnishes and the CPP-ACP toothpaste was observed to promote the remineralization of both mild and moderate MIH defects.

Humans↗

In vitro enamel remineralization at orthodontic band margins cemented with glass ionomer cement.

Demineralization adjacent to orthodontic bands remains a clinical concern. The release of fluoride from glass ionomer cement has been shown to inhibit demineralization. The purpose of this study was to examine the remineralization effects of a glass ionomer cement adjacent to orthodontic bands. Forty extracted molars were painted with an acid-protective varnish, excluding a 2 x 6 mm window on the buccal surfaces. Artificial caries-like lesions were created in the exposed enamel with an acidified gel. Half the lesion on each tooth was then painted with an acid-protective varnish. An orthodontic band was cemented with a glass ionomer cement or zinc phosphate cement, the gingival margin of the band located at the level of the artificial carious lesion. The teeth were placed in separate closed environments of a nonfluoridated artificial saliva for 3 months. The teeth were then sectioned and photographed with polarized light microscopy in imbibition medias of water and Thoulet's (R.I.: 1.41, 1.47), representing a minimum of 5%, 10%, and 25% pore volume, respectively. With a sonic digitizer, the area of the body of the lesion was measured in each imbibition media, comparing the maintained varnished out lesions to the lesions exposed to the cement. Results demonstrated a statistically significant reduction in the body of the lesions (p < 0.005) for those exposed to the glass ionomer cement.

Dental Caries↗

Biological factors in dental caries: role of saliva and dental plaque in the dynamic process of demineralization and remineralization (part 1).

Dental caries is a complex disease process that afflicts a large proportion of the world's population, regardless of gender, age and ethnicity, although it does tend to affect more indivduals with a low socioeconomic status to a greater extent. The process of dental caries is dependent upon biological factors that are present within the saliva and dental plaque. There are many different agents within saliva and plaque that serve to protect the tooth surface against caries development. Salivary flow rate, buffering capacity, antimicrobial activity, microorganism aggregation and clearance from the oral cavity, immune surveillance, and calcium phosphate binding proteins all interact to inhibit or reverse demineralization of exposed tooth surfaces. Cariogenic bacteria levels within the saliva and plaque determine whether caries will occur or not, and the concentration in saliva and plaque are intimately related to the type of carbohydrate ingestion and the frequency of ingestion, as well as the oral hygiene practiced by the individual.

Dental Caries↗

Remineralization of etched dentin.

To simulate a clinical cavity wall with the inner layer of carious dentin preserved, an artificially demineralized layer was made on the axial wall class V cavities of monkeys by a quick decalcifying solution. The cavities were then restored with a chemically adhesive composite resin after etching with a 40% phosphoric acid jelly etchant. Comparison of the Knoop hardness, the calcium concentration, and the scanning electron microscope observation of the dentin at the axial wall revealed that the dentin, demineralized by etching, was completely remineralized after 4 months.

Acid Etching, Dental↗

Effect of 2% plain and fluoridated milk on root surface caries in vitro.

The caries-protective effect of milk and fluoridated milk on enamel has been reported; however, few data are available concerning the role of milk and/or fluoridated milk on root surfaces. The aim of this study was to compare the effect of plain and fluoridated milk on root surface caries. Artificial root surface lesions were created, sectioned and analyzed using polarized light microscopy (PLM) and microradiography (MRG). The sections were covered except for the original surface and assigned to a treatment group. The samples were immersed in 2% plain milk or fluoridated milk for 40 hours and re-evaluated. Changes were measured and mean differences were compared with an unpaired t-test. Both techniques revealed a reduction in lesion depth for each milk group; however, a significantly greater reduction (p<0.05) was observed with the fluoridated milk. These results suggest that fluoridated milk may have a beneficial effect on the remineralization of root surface caries.

Animals↗