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In vivo caries models--mechanisms for caries initiation and arrestment.

The effects of intra-oral mechanical forces on caries initiation, progression, and arrestment are evaluated by examination of different in vivo caries models. The models are grouped in four categories: (1) a population study, (2) short-term clinical trials, (3) clinical experiments, and (4) controlled clinical observations. Taken together, these in vivo studies convincingly demonstrate that partial or total elimination of the intra-oral mechanical forces operating during mastication or toothbrushing leads to evolution of cariogenic plaque, resulting in localized carious enamel dissolution. In addition, they show that re-exposure to the partly or totally eliminated mechanical forces not only arrests further lesion progression, but also results in partial lesion regression. The data from in vivo caries studies also show that the clinical and structural changes associated with lesion arrestment or partial regression are not related to any salivary repair mechanism, but are solely the result of mechanical removal of the cariogenic biomass which is physically interrelated with the eroded surface of the active, dull-whitish enamel lesion. No indications of superficial mineral deposition or "blocking" of the external intercrystalline spaces are seen in the surface layer of lesions arrested in vivo. For this reason, the conventional usage of the terminology 'remineralization' is considered absolutely misleading when used to describe the mechanisms responsible for the arrest of lesion progression in vivo.

Clinical Trials as Topic↗

Fluoride uptake, retention, and remineralization efficacy of a highly concentrated fluoride solution on enamel lesions in situ.

Repeated topical application of concentrated fluorides is known to reduce caries. Little is known about fluoride retention and remineralization in incipient caries lesions following a single application. We investigated fluoride and the remineralization kinetics of a single application of elmex fluid (GABA International AG, Münchenstein, Switzerland; 10,000 ppm F) in initial enamel lesions. In this double-blind, placebo-controlled, randomized, crossover in situ study that conformed to good clinical practice, volunteers received intra-oral removable appliances carrying demineralized enamel samples after application of elmex fluid or placebo. After 5 min, 1, 2, 3, and 4 weeks in situ, KOH-soluble fluoride (KOHF), structurally bound fluoride (SBF), mineral gain, and lesion depth reduction were measured. Elmex fluid promoted higher KOHF and SBF at all times, decreased KOHF with time, increased SBF up to 3 weeks, and registered a higher mineral gain than placebo. Volunteers with higher stimulated salivary flow rates had lower fluoride uptake, but higher mineral gain. In conclusion, a single application of highly concentrated fluoride solution increases remineralization.

Adolescent↗

The influence of a matrix metalloproteinase on the remineralization of artificially demineralized dentin.

PURPOSE: This investigation aimed at determining whether a collagenolytic matrix metalloproteinase (MMP-8) present in the oral fluids might attack demineralized dentin and thereby affect the potential for remineralization. MATERIALS AND METHODS: Dentin specimens from freshly extracted third molars were demineralized for 14 days in vitro and then remineralized for 7 days in the presence or absence of activated MMP-8. Following treatment the mineral level of the specimens was assessed from the outer dentinal surface using Electron Probe Micro Analysis (EPMA). RESULTS: The findings suggested that the addition of activated MMP-8 to the remineralizing solution led to reduced remineralization in the surface of the dentin. There appeared to be no effect on the remineralization from any dentin bound MMP activity. CONCLUSION: It appears that MMP-8 may negatively influence the remineralization of demineralized dentin.

Acetates↗

Influence of fluoride on in vitro remineralization of artificial subsurface lesions determined with a sandwich technique.

Remineralization experiments of bovine enamel with subsurface lesions were carried out with the use of a sandwich technique. The remineralizing solutions contained no or 2 ppm fluoride added to the solution. The amount of remineralization was determined after remineralization periods up to 324 by means of quantitative microradiography. After 84 h the group with fluoride showed significantly (p less than 0.05) less remineralization than the nonfluoride group. This inhibition of fluoride on the remineralization may be explained by the inhibiting effect that fluoride at certain concentrations has on the crystal growth of apatites.

Animals↗

Remineralization of artificial caries-like lesions in human enamel in situ by chewing sorbitol gum.

The objective of the study was to determine quantitatively the effect on the potential for in situ remineralization of artificial caries-like lesions in human enamel when sugar-free gum containing mainly sorbitol as sweetener was chewed after meals and snacks. Artificial white-spot lesions were created in extracted human premolars and divided into three parts. One part was used as reference and the other two worn consecutively for two 21-day periods by 10 volunteers in a cast silver band cemented on lower molar teeth and covered with gauze to promote plaque formation. During the experimental periods, the subjects used fluoridated toothpaste twice daily, and consumed three meals (breakfast, lunch, and dinner) and two snacks (selected from chocolate bar, raisins, chocolate wafer, and iced cupcake). Sorbitol gum was chewed for 20 min immediately after each meal or snack during one of the experimental periods. The three parts of the enamel lesions were then sectioned (congruent to 80 microns) and examined together by means of quantitative microradiography and by polarized light microscopy. All estimates of mineral content indicated that significant remineralization occurred and was approximately doubled with gum-chewing. It is suggested that sorbitol gum stimulates salivation, which is responsible for the significantly enhanced remineralization, thus contributing to a therapeutic, caries-preventive effect. Because the gum was chewed immediately after meals and snacks, inhibition of demineralization may also have occurred.

Adult↗

Soluble calcium/SMFP dentifrice: effect on enamel fluoride uptake and remineralization.

PURPOSE: To evaluate in situ the effect of soluble calcium on fluoride uptake and remineralization by human dental enamel from a sodium monofluorophosphate (SMFP) dentifrice. MATERIALS AND METHODS: Eight volunteers took part in this cross-over, double-blind design study performed in three phases of 45 days. During each phase of the experiment, the subjects wore an acrylic resin appliance containing four blocks of human dental enamel with artificial caries to evaluate fluoride uptake and remineralization by three dentifrices: SMFP containing CaCl2, SMFP, and placebo. After each phase, the enamel blocks were removed and the total fluoride uptake (ppm F) and microhardness (Knoop) were measured. Statistical analyses (ANOVA and Turkey's test) were done. RESULTS: There was no statistically significant difference between SMFP-CaCl2 and SMFP treatments regarding the amount of fluoride and microhardness data of treated enamel blocks as well as their areas under the curves, although both differed significantly (P < 0.05) when they were compared with placebo. However, the SMFP-CaCl2 treatment demonstrated the highest values on fluoride uptake and microhardness data. Furthermore, the microhardness data demonstrated 50% and 40% of remineralization for SMFP-CaCl2 and SMFP dentifrices, respectively.

Analysis of Variance↗

A paradigm shift in the treatment of caries.

This paper presents a shift in the paradigm for treating dental caries from a surgical to a medical approach. Two key points are essential to understanding caries. First, caries is a bacterial infection caused by specific bacteria. Second, before cavitation, caries is a reversible multifactoral process. Two features, caries activity and caries risk, provide a diagnosis and prognosis leading to a treatment plan. Patients receive treatment using a medical model consisting of four steps: bacterial control; reduction of risk levels for at-risk patients; reversal of active sites by remineralization; and follow-up and maintenance.

Anti-Infective Agents, Local↗

In situ evaluation of different remineralization periods to decrease brushing abrasion of demineralized enamel.

The aim of the present in situ study was to evaluate the effect of different periods of intraoral remineralization to decrease the susceptibility of previously demineralized enamel against toothbrushing abrasion. Six human enamel specimens (A-F) were recessed in the buccal aspects of each of eight intraoral appliances which were worn for 21 days by 8 panelists. Demineralization of the samples was performed twice a day extraorally in the acidic beverage Sprite Light for 90 s. Subsequently, the enamel specimens were brushed at different times. Specimen A was brushed immediately after the demineralization. The remaining samples B-E were brushed after the intraoral appliances had been worn for various periods of remineralization: specimen B, 10 min; C, 20 min; D, 30 min and E, 60 min, respectively. Specimen F was only demineralized and remineralized, but not brushed. After 21 days, enamel wear was measured with a laser profilometer. The following values (mean +/- standard deviation) were obtained: specimen A, 6.78+/-2.71 microm; B, 5.47+/-3.39 microm; C, 6.06+/-3.18 microm; D, 5.43+/-2.58 microm; E 4.78+/-2.57 microm, and F 0.66+/-1.11 micro;m. Analysis of variance revealed a significant influence of remineralization period on abrasive wear. However, even after a remineralization period of 60 min the wear was significantly increased as compared to the demineralized, but not brushed control. It is concluded that (1) abrasion resistance of softened enamel increases with remineralization period and (2) at least 60 min should elapse before toothbrushing after an erosive attack.

Amines↗

A study of the ability of an in situ remineralization model to differentiate between the effects of two fluoride dentifrices that produced significantly different clinical caries results.

The purpose of this study was to determine whether an in situ remineralization model was able to show a difference in the effects of two dentifrices of different fluoride concentrations and significantly different clinical efficacies. Three dentifrices were tested in a double-blind, cross-over study design. The products contained 0 ppm F, 250 ppm F, or 1000 ppm F from sodium fluoride and were formulated with a silica base according to the formulations used in a human caries trial (Koch et al., 1990). Nineteen subjects each carried three or four thin sections of enamel in their partial dentures. The thin sections, containing artificial caries lesions, were covered with a steel mesh to provide space for plaque formation and then brushed in situ three times daily with the dentifrices. Following the two-week treatment periods, the specimens were removed from the dentures and analyzed for changes in mineral content. The findings showed that the placebo dentifrice (0 ppm F) resulted in 56.8 +/- 74.3% demineralization, the 250-ppm-F dentifrice produced 12.9 +/- 41.3% demineralization, thereby showing partial caries protection, and the 1000-ppm-F dentifrice produced 17.3 +/- 32.1% remineralization. Linear regression analysis showed that the percent remineralization was significantly related to the fluoride concentration in the dentifrice (p less than 0.001). The 250-ppm-F dentifrice was also significantly less effective than the 100-ppm-F dentifrice (p = 0.04, one-tailed Fisher Protected LSD test). These findings are in accord with the human caries trial and support the use of the present in situ remineralization model for prediction of the anticaries efficacy of fluoride dentifrice products.

Adult↗

Remineralized dentin lesions induced by glass ionomer demonstrate increased resistance to subsequent acid challenge.

OBJECTIVE: The purpose of this study was to test the hypothesis that remineralized dentin lesions induced by glass ionomer are less vulnerable to subsequent acid challenge. METHOD AND MATERIALS: Baseline demineralized (BDe) lesions were created in 50 bovine dentin slices immersed for 3 weeks in acid solution. A resin-modified glass ionomer (RMGI) was applied to the specimens before immersion in salivalike solution to allow remineralization (Re) for 2 weeks (V2W) or 6 weeks (V6W). Resin-modifed glass ionomer was coated to block further ion release before a 3-week immersion in acid solution as a second demineralization (SDe). Control groups (C2W and C6W) were done simultaneously without RMGI. In another group, RMGI was left undisturbed (V2W+) to allow continuous ion release during SDe. Mineral content was converted from microradiographs, after BDe, Re, and SDe steps. Changes in mineral content were calculated and compared between groups. RESULTS: The remineralized surface zone was maintained after SDe, but the lesion body became deeper. Percentage of mineral loss from SDe was not significantly different between V2W and C2W (t test, P > .05). When remineralization was extended to 6 weeks, V6W showed significantly less mineral loss than C6W. Mineral loss was lowest when RMGI was not blocked (V2W+). CONCLUSION: Dentin lesions remineralized in the presence of RMGI maintained the highly mineralized surface zone when subjected to second demineralization. However, the remineralized surface zone could not prevent advance of the lesion body. The subsequent demineralization was markedly reduced by extending the remineralization period or by leaving the RMGI undisturbed to resume ion release.

Acids↗

Chemical and structural challenges in remineralization of dental enamel lesions.

The aim of the present paper was to identify some chemical and structural factors which may prevent a full remineralization of caries lesions and to study whether it is possible to overcome such obstacles. Samples of powdered enamel apatite were equilibrated with solutions metastably supersaturated with respect to enamel hydroxyapatite and fluorapatite. After 10 min and 60 min of equilibration at 20 degrees C the suspensions were centrifuged and the calcium and phosphate concentrations and the pH were determined in the supernatant. In parallel studies, 50 75-microns-thick sections of 27 fluorotic teeth of a severity of 5-7 according to Thylstrup and Fejerskov's classification were examined by microradiography and in polarized light using distilled water, and Thoulett's media or seen dry in air. Five obstacles inhibiting remineralization were identified: 1) Although remineralizing solutions or saliva are supersaturated with respect to enamel apatite the total amount of calcium and phosphate dissolved in it is small, so that after precipitation of the dissolved mineral only 1/20,000-1/30,000 of the volume of the mineralizing solution is occupied by mineral. 2) The concentration gradients from the mineralizing solution into the enamel is small, which indicates a slow diffusion into and out of the lesion. 3) The uptake of calcium and phosphate by the enamel apatite crystals is so rapid that the aqueous phase within the pores can be presumed to be only marginally supersaturated in the deeper parts of the lesion. 4) The surface layer of the enamel lesions was found to be a serious obstacle to remineralization so that a subsurface area remains hypomineralized after exposure to salivary remineralization even for a lifetime. 5) Nucleation of new apatite crystals to substitute lost crystals is an unsolved problem.

Apatites↗

Effect of dentifrice containing fluoride and/or baking soda on enamel demineralization/remineralization: an in situ study.

The additive effect of baking soda on the anticariogenic effect of fluoride dentifrice is not well established. To evaluate it, a crossover in situ study was done in three phases of 28 days. Volunteers, using acrylic palatal appliances containing four human enamel blocks, two sound (to evaluate demineralization) and two with artificial caries lesions (to evaluate remineralization), took part in this study. During each phase, 10% sucrose solution was dripped (3 times a day) only onto the sound blocks. After 10 min, a slurry of placebo, fluoride (F) or fluoride and baking soda (F+NaHCO(3)) dentifrice was dripped onto all enamel blocks. The results showed a higher F concentration in dental plaque formed during treatment with F+NaHCO(3) than placebo (p<0.05), but the difference related to F dentifrice was not significant. The enamel demineralization was lower, and remineralization was greater, after treatment with F+NaHCO(3) than placebo (p<0.05), but the difference related to F dentifrice was not significant. The data suggest that baking soda neither improves nor impairs the effect of F dentifrice on reduction of demineralization and enhancement of remineralization of enamel.

Buffers↗

Effect of a solution with high fluoride concentration on remineralization of shallow and deep root surface caries in vitro.

A root dentin single-section model was developed to compare the efficacy of different fluoride treatments on the remineralization of lesions. Shallow ( approximately 170 microm) and deep ( approximately 400 microm) lesions were produced in acetic acid buffer solutions (pH 5.0) with 0.1- and 0.5-ppm fluoride for 3 days and 2 weeks, respectively. Next, the sections were pH-cycled for 4 weeks. Following the pH cycling, all sections were first remineralized for 5 weeks, then subjected to 10 days of demineralization. The treatments were (1) no treatment (control), (2) daily 1,450-ppm NaF toothpaste, (3) weekly 4,000-ppm fluoride solution, (4) a combination of treatments 2 and 3. In the shallow lesions, the 4,000-ppm fluoride solution and the combination treatment enhanced mineral deposition at the lesion front, producing a second, slightly hyperremineralized layer. Similarly, in the deep lesions a second remineralized layer was detected. In all lesions, the fluoride solution treatment showed significantly more remineralization compared to the control and the toothpaste treatment (p<0.05), most of which was formed during the pH cycling. In the demineralization period, the control showed significantly higher mineral loss than all other treatments. These results indicate that a 4,000-ppm fluoride solution might be effective for remineralization of root dentin lesions even over 400 microm depth.

Acetates↗

Caries-preventive agents induce remineralization of dentin in a biofilm model.

The aim was to study remineralization in dentin underneath a biofilm. This was done in a constant depth film fermentor (CDFF) which was modified so that two treatments can be applied simultaneously in one experiment. Forty-five Streptococcus mutans biofilms were grown in grooves in dentin. Growth medium (3.7 g/l BHI, 1.5 mM calcium and 25 mM PIPES) was administered alternately with 2% sucrose pulsing 4 x 30 min/day. Fluoride at 135 ppm as NaF only or in a mixture with 0.2% chlorhexidine was applied for 2 x 5 min/day. The treatments started 5 days after inoculation and lasted 15 days. Five specimens per group were removed at various time points. The biofilms were checked for viability (by plating) and acid content (by capillary electrophoresis). The dentin specimens were analysed for mineral loss and lesion depth (by transversal microradiography). Fluoride treatment had no effect on the viability but reduced lactic acid production by 75%. The mixture treatment reduced the viability by 80% and the lactic acid content by 93% on the first day and later reduced the two parameters to below the detection limits. Significant differences in changes in mineral loss and lesion depth were observed between the treatment groups. Partial remineralization but deeper lesions were observed in the fluoride group, while nearly complete remineralization was seen in the mixture group. In conclusion, the CDFF S. mutans biofilm model can be used as a de- and remineralization biofilm model, and the split mode is particularly suitable for testing caries-preventive agents.

Animals↗

Fluoride and mineral redeposition in outermost layers of bovine enamel during surface softening.

Mineral and fluoride concentration changes in the outermost layers of bovine enamel (depth less than 1 micron) were measured after demineralization in unbuffered hydroxyethylcellulose gels of pH = 5.4 with an intrinsic fluoride concentration of about 0.02 ppm. A combination of two nuclear analytical techniques, Rutherford backscattering spectrometry (RBS) and proton-induced gamma ray emission spectrometry (PIGE) was applied to determine the Ca/P molar ratios and F depth profiles, respectively. When compared to deeper layers, a reduced loss of mineral content is observed for the depth range of about 0-0.1 micron corresponding well with a F concentration increase from about 500 to about 5,000 ppm in the same range. These findings are interpreted as a fluoride-induced partial remineralization of the superficial surface layer during an overall demineralization process.

Animals↗

In situ remineralization of subsurface enamel lesion after the use of a fluoride chewing gum.

In situ remineralization of early enamel lesions by a fluoride chewing gum was studied. Human enamel specimens with subsurface lesions were mounted in removable lower appliances for 6 adults. Subjects used a F-free dentifrice 3x/day and chewed five sticks/day for the F gum group (0.1 mg F/stick) or five sticks of sugarless gum. No gum was chewed for controls. Surface microhardness was performed on: (1) sound enamel; (2) lesions; (3) after intraoral exposure, and (4) after acid-resistance testing (ART). Separate specimens were etched and measured for F uptake and image analyses on microradiographs were performed for all regimens. delta Z values were calculated and converted to percent of mineralization. Values for F gum were significantly higher (p > 0.05) than non-F gum and controls for ART, percent remineralization, and F uptake up to 70 microns depth.

Adult↗

Dose related remineralization using intraoral fluoride-releasing devices in situ.

Dose-related remineralization of enamel lesions was studied in situ. Enamel specimens with subsurface lesions were mounted on lower molars bilaterally and exposed to five regimens: (A) control with a F-free dentifrice three times daily; (B) NaF dentifrice brushed three times daily; (C) fluoride-releasing device (FRD; 0.07 mg F/day release rate) plus F-free dentifrice; (D) FRD (0.232 mg F/day) plus F-free dentifrice, F/day and (E) FRD (0.232 mg/F/day) plus NaF dentifrice. Resting and stimulated saliva samples were obtained for salivary flow and F content determinations. Twenty-eight subjects participated in this double-blind, random-assignment crossover study. After 50 days the enamel specimens were recovered, sectioned, microradiographed, and scanned with image analysis. Regimens D and E demonstrated approximately 30% additional remineralization as compared with to controls, and regimens B and C showed 13.7 and 10.5% increased remineralization as compared with to controls. The increased remineralization of the regimens generally paralleled the increase in salivary F and clearly demonstrated a dose-related response which was some threefold in relation to release rate, salivary F, and percent remineralization.

Adult↗