Caries-like Lesions Research Articles

Comparing two quantitative methods for studying remineralization of artificial caries

Objectives To compare the detection of changes before and after remineralization of artificial enamel and dentin caries by microCT scanning, polarized light microscopy (PLM) and transverse microradiography (TMR). Methods Fourteen extracted premolars were cut into tooth blocks and painted with an acid-resistant varnish leaving one enamel and one dentin surface exposed. The tooth blocks were immersed into demineralizing solution for 4 days to produce artificial caries-like lesions and scanned by microCT. Then the 14 tooth blocks were randomly allocated into two groups. Seven tooth blocks in Group I were cut longitudinally through the exposed surface into 100–150 μm thick sections and microradiographs were taken. The other seven tooth blocks in Group II were left intact. All the tooth blocks and sections were then immersed into remineralizing solution for 5 days. PLM and TMR of the tooth sections in Group I were taken again. Depth of the lesion on the TMR was measured. Tooth blocks in Group II were scanned by microCT. Results Mean lesion depth in Group I reduced by 13.0% and 8.2% after remineralization for enamel and dentin, respectively (paired t-test, P < 0.001). In Group II, linear attenuation coefficient (LAC) of the region of interest (ROI) increased by 11.1% and 23.8% after remineralization for enamel and dentin lesions, respectively (paired t-test, P < 0.001). Conclusion Both microCT and microradiography are able to detect a change of similar magnitude in the artificial caries lesions after remineralization. MicroCT may be used to substitute TMR and PLM in in vitro studies about caries.

Automated analysis of lesion depth and integrated reflectivity in PS‐OCT scans of tooth demineralization

Several studies have demonstrated that polarization sensitive optical coherence tomography (PS-OCT) can be used to nondestructively measure the severity of subsurface demineralization in enamel and dentin, track lesion progression over time and measure remineralization. The purpose of this study was to develop methods for the automated assessment of the depth and severity of demineralization in PS-OCT scans. Subsurface caries-like lesions of increasing depth and severity were produced in adjoining windows on 10 bovine enamel samples via exposure to demineralization for periods of 1-4 days. PS-OCT scans were acquired for each sample and analyzed using various methods to calculate the lesion depth and severity. Edge detection algorithms were most successful for measurement of the lesion depth for improved assessment of lesion severity. Edge-finding algorithms were able to detect significant differences (P<0.05) in the lesion depth and severity between each of the periods of demineralization and sound enamel. The lesion depth and mineral loss were also measured with polarized light microscopy and transverse microradiography after sectioning the teeth for comparison. This study demonstrates that the depth and severity of early lesions can be calculated automatically for rapid analysis of PS-OCT images.

Effect of drying on the reproducibility of DIAGNOdent to detect caries-like lesions

The aim of this study was to verify the drying effect on the reproducibility of DIAGNOdent (Dd) devices to detect caries-like lesions. Three areas were created in each of the 34 bovine incisors: sound (S), demineralized (DE) and remineralized (RE). One examiner measured each area with two Dd devices (denominated X and Y), twice under humid, and twice under dry condition. Intra-rater agreement according each device and inter-device agreement were estimated by kappa statistics (k). Intra-rater agreement for device Y was substantial under humid (κ (DE)=0.68 and κ (RE+S)=0.68) and dry condition (k DE=0.64 and k RE+S=0.67). For device X, it was substantial under humid condition (k DE=0.57 and k RE+S=0.49), and it was almost perfect after air drying (κ (DE)=1.0 and κ (RE+S)=1.0). Inter-device agreement was slight (k =0.17) under humid condition, and it was substantial under dry condition (k =0.62). As reproducibility increased under dry condition, drying is advised to detect caries-like lesions on free smooth surfaces when different devices are used.

Demineralization Depth Using QLF and a Novel Image Processing Software

Quantitative Light-Induced fluorescence (QLF) has been widely used to detect tooth demineralization indicated by fluorescence loss with respect to surrounding sound enamel. The correlation between fluorescence loss and demineralization depth is not fully understood. The purpose of this project was to study this correlation to estimate demineralization depth. Extracted teeth were collected. Artificial caries-like lesions were created and imaged with QLF. Novel image processing software was developed to measure the largest percent of fluorescence loss in the region of interest. All teeth were then sectioned and imaged by polarized light microscopy. The largest depth of demineralization was measured by NIH ImageJ software. The statistical linear regression method was applied to analyze these data. The linear regression model was Y = 0.32X + 0.17, where X was the percent loss of fluorescence and Y was the depth of demineralization. The correlation coefficient was 0.9696. The two-tailed t-test for coefficient was 7.93, indicating the P-value = .0014. The F test for the entire model was 62.86, which shows the P-value = .0013. The results indicated statistically significant linear correlation between the percent loss of fluorescence and depth of the enamel demineralization.

Artificial methods of dentine caries induction: A hardness and morphological comparative study

Objective To assess the ability of two chemical and a microbiological methods to produce dentine caries lesions resembling naturally developed dentine caries lesions. Design Forty sound second primary molars were divided into four experimental groups according to the method to produce artificial caries lesions: (1) sound (negative control); (2) acidified gel; (3) pH-cycling; and (4) microbiological, all for 14 days. Ten second primary molars presenting natural dentine caries lesions comprised the (5) positive control group. After the artificial caries induction, all samples were longitudinally sectioned and polished in order to obtain Knoop microhardness values from 10 to 500 μm depth from the bottom of the cavities. Morphological analysis of the surfaces was carried out by SEM. Hardness data were compared among the five experimental groups using One-Way ANOVA and post hoc SNK's test. Results The hardness values of chemically created caries-like lesions did not differ from that of natural caries lesions on shallower depths. The results indicated that chemical caries induction methods promote a superficial demineralization and that pH-cycling is more effective than acidified gel. The former, produced a thicker layer of demineralization, with similar hardness values than natural lesions. Despite the microbiological method provided an excessive softness of the primary dentine, this method presented morphology more comparable to natural lesions. Conclusions pH-cycling is more appropriated to simulate a substrate that resembles affected caries dentine layer, after caries removal. The microbiological method seems more indicated to simulate a dentine caries lesion with an infected layer, previously to caries removal.

Cavity Preparation and Influence of Restorative Materials on the Prevention of Secondary Caries

This in vitro study evaluated the influence of cavity preparation using the Er:YAG laser and restorative materials containing fluoride on preventing caries lesions. It has been suggested that cavity preparation using the Er:YAG laser has a potential for improving resistance to secondary caries on enamel. Forty unerupted human third molars teeth were sectioned into 72 blocks of dental enamel and distributed into two groups to prepare cavities measuring (1.6 mm diameter) with diamond burs (DB) or Er:YAG laser (LA; 6 Hz, 300 mJ, 47 J/cm(2)). After that, each group was divided into three subgroups and restored with a glass-ionomer cement (GI), a resin-modified glass-ionomer (RM), or a composite resin (CR). Blocks were thermal cycled and submitted to a pH challenge to develop artificial caries-like lesions. Lesions were evaluated by Knoop microhardness test. An average of four indentations was used. Statistical analyses were performed by ANOVA followed by Tukey's test. The results (in Knoop hardness number) for DB cavity preparation were GI, 235.5 (+/-75.5); RM, 137.1 (+/-64.1); and CR, 39.3 (+/-26.5). For LA cavity preparation, the results were GI, 410.0 (+/-129.7); RM, 310.3 (+/-119.5); and CR, 96.4 (+/-57.4). There was less development of caries lesion around LA-prepared cavities than around the DB-prepared cavities; however, no synergistic cariostatic effect was observed between the Er:YAG laser and glass ionomer cement.

Potential agents to control enamel caries-like lesions

Objectives To assess whether pastes containing casein phosphopeptide–amorphous calcium phosphate (CPP–ACP) and calcium sodium phosphosilicate (CSP) control artificial caries lesion progression. Methods Enamel slabs of bovine teeth were embedded in polyester resin and had their enamel surfaces serially polished, and tested for surface microhardness (SMH, 25 g, 5 s). Incipient caries-like lesions were pre-formed and specimens were evaluated by microhardness test (SMH post-lesion) and randomly assigned to five treatment groups ( n = 15): (1) regular dentifrice (RE, 1,100 ppm F); (2) dentifrice with calcium sodium phosphosilicate (CSP); (3) amorphous calcium phosphate stabilized by casein phosphopeptide (CPP–ACP); (4) CPP–ACP with 900 ppm F (CPP–ACP + F) and (5) control group—unexposed to any remineralizing agent. Treatments were applied five times, after the de-remineralization period in the cariogenic challenges. Post-treatments SMH measurements were conducted (SMH post-treatment). Results ANOVA was applied for data evaluation and revealed a significant difference among the treatments ( p = 0.0161). Tukey's test was conducted and the percentage of mineral loss was calculated. Specimens exposed to CSP (7.1%), RE (6.7%) and CPP–ACP + F (3.8%) showed lower mineral loss than those that in the control group (−11.0%). CPP–ACP group (3.2%) differed from the control, CSP and RE groups. Conclusions Depending on the agent used, a remineralizing effect may be expected, which reflects in caries lesions progression.

Methods for calculating the severity of demineralization on tooth surfaces from PS-OCT scans.

Several studies have demonstrated that polarization sensitive optical coherence tomography (PS-OCT) can be used to nondestructively measure the severity of subsurface demineralization in enamel and dentin. The reflectivity in the polarization state orthogonal to the initial linear polarization incident on the tissue is low at sound tissues interfaces and high in demineralized areas that strongly scatter and depolarize the light. The purpose of this study was to develop improved algorithms for assessing the depth and severity of demineralization from PS-OCT scans for use with 2D and 3D tomographic images. Subsurface caries-like lesions of increasing depth and severity were produced in adjoining windows on ten bovine enamel samples by exposure to demineralization over periods of 1 to 4 days. Each sample also had a sound window to be used as a control. PS-OCT scans were acquired for each sample and analyzed using various methods to calculate the lesion depth and area. Algorithms were developed and used to automatically detect the lesion depth and area, and calculate the volume for improved assessment of lesion severity. Both fixed-depth and automatic edge-finding algorithms were able to detect significant differences between each of the days and sound enamel. The lesion depth and mineral loss were also measured with polarized light microscopy and transverse microradiography after sectioning the teeth. Mean lesion depths ranged from 40 to 100 μm. This demonstrates the edge-finding algorithm can be used to automatically determine the depth and severity of early lesions for the rapid analysis of PS-OCT images.

The Efficacy of Laser Fluorescence to Detect in Vitro Demineralization and Remineralization of Smooth Enamel Surfaces

The purpose of this study was to evaluate the efficacy of the laser fluorescence (LF) device in detecting in vitro demineralization and remineralization of smooth surface caries-like lesions. The early detection of smooth surface caries-like lesions is important to provide proper management of carious lesions, and allows monitoring of them over time. Also, some authors suggest that LF could be useful in monitoring the caries pathological process. Seventy-eight blocks of bovine teeth were obtained, and then submitted to artificial caries lesion induction and to a pH-cycling process. Superficial microhardness (SMH) and laser fluorescence analysis were performed at baseline, after induction of caries-like lesions, and after the pH-cycling regimen to promote remineralization. Friedman's and multiple comparison tests were performed for all variables. SMH analysis showed significant differences (p +/- 0.05) between baseline (286.77 +/- 1.49 Vickers hardness number [VHN] units), before (38.48 +/- 0.85 VHN), and after remineralization (131.93 +/- 2.63 VHN). Baseline values for LF were extremely low (2.71 +/- 0.05), and a statistically significant difference was observed only after remineralization (3.61 +/- 0.08), as demonstrated by the increase in LF values. The LF device did not show efficacy for monitoring in vitro demineralization and remineralization of smooth enamel surfaces.

Comparison of demineralisation rates in pre- and postnatal enamel and at the neonatal line

AimsThe neonatal line, which is an exaggerated incremental layer line, separates pre- and postnatal enamel. It has been suggested that this layer may be a barrier to the progress of a carious lesions. The objective was to measure the rate of demineralisation in pre- and postnatal enamel and within the neonatal layer using scanning microradiography (SMR). Permanent enamel and compressed permeable hydroxyapatite samples were used as controls. Methods and resultsEnamel specimens from deciduous incisors were cut into mesiodistal blocks of 2mm thickness without altering the labial surface and located within SMR cells. Permanent enamel and hydroxyapatite specimens were similarly prepared. Artificial caries-like lesions were created by exposing the specimens to 0.1moll−1 acetic acid (pH 4.0) within the SMR cells. SMR was used to measure the rate of mineral loss at 10 points either side of and at the neonatal line in the deciduous enamel, and 20 points across in the control specimens.The rate of demineralisation was almost the same in pre- and postnatal enamel ((6.0–8.0)×10−4gcm−2h−1), but much lower in the vicinity of the neonatal line (2.0×10−4gcm−2h−1). The rate of demineralisation was lower in permanent (5.0×10−4gcm−2h−1) than in deciduous enamel, and even lower in the permeable hydroxyapatite specimen (2.5×10−4gcm−2h−1). ConclusionsThis study showed no difference in the rate of demineralisation between pre- and postnatal enamel, but a reduced rate within the region that contained the plane of the neonatal line. This supports the hypothesis that the neonatal line may act as a barrier to the propagation of carious lesions.

Effect of strontium in combination with fluoride on enamel remineralisation in vitro

Previous studies showed that strontium (Sr) as well as fluoride (F) can enhance enamel remineralisation. The aim of this study was to evaluate the effects of Sr in combination with F on enamel remineralisation 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 remineralising solutions (1.5 mM CaCl 2, 0.9 mM KH 2PO 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 remineralisation 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. Remineralisation was significantly affected by F concentration ( p = 0.000). The participation of Sr resulted in a significant enhancement of remineralisation ( p < 0.001) with a synergistic effect of the Sr + F combination ( p < 0.01). It was concluded that while the remineralising process was affected by the concentration of F, there was also an interaction between F and Sr when they were used in conjunction.

The effect of casein phosphopeptide‐amorphous calcium phosphate on remineralization of artificial caries‐like lesions: an in vitro study

The aims of this study were to investigate the efficacy of CPP-ACP containing Tooth Mousse on the remineralization of enamel lesions and to compare its efficacy to that of a fluoride-containing toothpaste. Permanent teeth were placed in demineralizing solution for 96 hours to produce artificial caries-like lesions 120-200 microm in depth. They were sectioned into 100-150 microm thick samples and randomly assigned to five groups: for Group A, a fluoridated toothpaste (1100 ppm) was used as a positive control and in Group B, a non-fluoridated toothpaste was used as a negative control. Tooth Mousse containing CPP-ACP was tested by three different means: as a toothpaste (Group C); as a topical coating (Group D); and (Group E) as a topical coating after treating the sections with the same fluoridated toothpaste as in Group A. The lesion depth decreased significantly by 7 per cent in Group A, 10.1 per cent in Groups C and D, and 13.1 per cent in Group E (Paired t- test, p < 0.05), while in Group B the lesion depth increased significantly by 23 per cent. Based on the data obtained, CPP-ACP containing Tooth Mousse remineralized initial enamel lesions and it showed a higher remineralizing potential when applied as a topical coating after the use of a fluoridated toothpaste.

Confocal laser scanning microscopic analysis of the depth of dentin caries-like lesions in primary and permanent teeth

This study analyzed comparatively, by confocal laser scanning microscopy (CLSM), the depth of caries-like lesions produced by biological and chemical artificial models in permanent and primary dentin. Six primary molars and six premolars were used. The occlusal enamel was removed and a nail polish layer was applied on the specimens, except for a 4 x 2 mm area on dentin surface. Half of specimens were immersed in acid gel for 14 days (chemical model) and the other half was immersed in BHI broth with S. mutans for 14 days (biological model). After development of artificial caries, the crowns were longitudinally sectioned on the center of the carious lesion. Three measurements of carious dentin depth were made in each specimen by CLSM. Measurements depths were compared between the caries models and between tooth types by one-way ANOVA and Tukey test (alpha=5%). For permanent teeth, the biological model showed significantly higher (p<0.05) caries depth values than the chemical model. For primary teeth, no statistically significant difference (p>0.05) was found between the caries models. The artificial caries model influenced caries depth only in permanent teeth. There was no difference in carious dentin depth between permanent and primary teeth, regardless of the artificial caries model.

Anticaries potential of a fluoride mouthrinse evaluated in vitro by validated protocols

This blind and randomized study tested in vitro, using validated protocols, the anticaries potential of an experimental fluoride mouthrinse. One-hundred enamel slabs, half sound and half with caries-like lesions (carious), all with known surface microhardness (SMH), were submitted to 3 treatment groups: A) a placebo mouthrinse (negative control); B) a positive control mouthrinse containing 0.05% NaF; and C) an experimental formulation containing 0.05% NaF and cetylpyridinium chloride as an antibacterial substance. To evaluate the formation of F products on enamel, sound (n=10) and carious (n=10) slabs were treated with the formulations during 10 min and loosely and firmly-bound F formed in enamel were determined after extraction with alkali and acid, respectively. To evaluate the inhibition of enamel demineralization, sound enamel slabs (n=10) were treated with the mouthrinse formulations 2x/day during 1 min and subjected to a pH-cycling regimen simulating a cariogenic challenge (demineralization). To evaluate enamel remineralization, the carious slabs (n=10) were submitted to the treatments 3x/day and subjected to a pH-cycling model simulating a remineralizing condition. After 8 days, enamel SMH was determined again and the percentage of SMH loss or SMH recovery was calculated for the sound and carious slabs, respectively. The experimental formulation was superior to the negative control (p<0.05) and equivalent to the positive control (p>0.05) in the formation of F products in enamel, and in the inhibition of enamel demineralization and enhancement of remineralization. These data suggest that the tested experimental fluoride mouthrinse has anticaries potential.

The Inhibitory Effect of MI Paste, Fluoride and a Combination of Both on the Progression of Artificial Caries-like Lesions in Enamel

This in-vitro study evaluated the inhibition of demineralization in enamel sections produced by MI paste, fluoride and a combination of both, compared to artificial saliva and NaF 5000 ppm in a caries progression pH-cycling model. Twenty-one teeth were demineralized to create subsurface enamel lesions (approximately 200 microns in depth). The teeth were sectioned and characterized using polarized-light-microscopy (PLM). A single section from each lesion was assigned to a treatment group: Artificial saliva, NaF 5000 ppm (Prevident, Colgate), MI paste (Recaldent, GC America Inc), NaF 1100 ppm (Crest, Procter & Gamble) and NaF 1100 ppm plus MI paste. The sections were covered with varnish except for an exposed window on the external surface of the lesion and placed in a six-day pH-cycling model with two daily treatment applications of two minutes each. The sections were characterized by PLM, and the lesion areas were measured using a digital image analysis system. Based on a paired-sample t-test, significant differences (p < .05) in percentage of change in lesion size were found between the high fluoride group and all the other groups. No significant difference was found between the artificial saliva and MI paste group, neither was there any significant difference between the NaF 1100 ppm, the combined application group or the MI paste group alone. In conclusion, the higher concentration of NaF (5000 ppm) reduced lesion progression to the greatest extent. The MI paste group did not show any effect on the inhibition of lesion progression. Further studies on the preventive effect and longer treatment applications are recommended.

Effects of Ozone and Sodium Hypochlorite on Caries-Like Lesions in Dentin

The hypothesis that ozone promotes remineralization of dentinal lesions was tested in vitro. Artificial caries-like lesions in dentin were treated with ozone gas, with another potent oxidizer (sodium hypochlorite, NaOCl, 10%) or with water. The specimens were then remineralized and subsequently demineralized again. Mineral content was assessed by transverse microradiography. NaOCl-treated samples showed damaged surface and, after being remineralized, demineralized significantly more than water- or ozone-treated groups. No difference was found between ozone and water groups. The exposure to ozone had thus no effect on remineralization and subsequent demineralization of remineralized dentinal lesions.

Effect of a CPP-ACP agent on the demineralization and remineralization of dentine in vitro

Objectives The aim of this study was to determine in vitro the effect of a commercial paste based on CPP-ACP complex on the demineralization of sound human dentine and on remineralization potential of artificial caries-like lesions formed on dentine surfaces. Methods Forty dentine specimens were prepared with hard tissue microtome. The specimens were divided in four groups the A, B, C and D ( n = 10). The specimen surfaces were subjected to surface analysis by Fourier transformance micro multiple internal reflectance infrared spectroscopy (micro MIR-FTIR). Tooth mousse was applied on surface specimens of A group, while no agent were applied on the specimens of B group. Afterwards, groups A, B, C and D were immersed in demineralization solution for 7 days. Afterwards, the surfaces were subjected to micro MIR-FTIR analysis and the mineral to matrix ratio was used to assess the extent of dentin demineralization (DM). Tooth mousse was applied on specimens of group C, while no agent was applied on specimens of group D. The groups C and D immersed in artificial saliva for 7 days and were subjected to analysis by micro MIR-FTIR and the mineral to matrix ratio was used to assess the extent of dentin remineralization (RM). Results Group A showed significant lower %DM in comparison to group B. Group C resulted in a significant higher %RM compared to group D. Conclusions The presence of agent CPP-ACP on dentine surfaces provoked lower demineralization and higher remineralization in comparison with the dentine surfaces without agent.

An in vitro microbial model for producing caries-like lesions on enamel

This study aimed to develop a low cost in vitro viable microbiological model to produce biofilms to be used in dental researches. Single and multi-species biofilms of S. mutans, S. sobrinus, S. mitis, S. salivarius, S. cricetus and S. sanguinis were grown on bovine enamel slabs during 10 days, in a sterile brain-heart infusion broth, containing 5% sucrose and incubated at 37°C in an atmosphere of 10% CO2. The slabs were transferred to a fresh medium at every 6, 12 or 24 hours. After the experimental period, enamel volume percent mineral was determined by cross-sectional microhardness. Caries-like lesions were found in all bacterial groups when compared with the control group. No statistical significant differences were found between S. mutans and S. sobrinus with respect of their cariogenicity or among the periods of medium change. However, it was found a statistical significant difference among the cariogenicity of S. salivarius and S. sanguinis (ANOVA followed by Tukey test). This model has successfully developed caries-like lesion on enamel and the medium can be changed at every 24 hours utilizing either S. mutans or S. sobrinus.

Self-assembling Peptide Scaffolds Promote Enamel Remineralization

Rationally designed beta-sheet-forming peptides that spontaneously form three-dimensional fibrillar scaffolds in response to specific environmental triggers may potentially be used in skeletal tissue engineering, including the treatment/prevention of dental caries, via bioactive surface groups. We hypothesized that infiltration of caries lesions with monomeric low-viscosity peptide solutions would be followed by in situ polymerization triggered by conditions of pH and ionic strength, providing a biomimetic scaffold capable of hydroxyapatite nucleation, promoting repair. Our aim was to determine the effect of an anionic peptide applied to caries-like lesions in human dental enamel under simulated intra-oral conditions of pH cycling. Peptide treatment significantly increased net mineral gain by the lesions, due to both increased remineralization and inhibition of demineralization over a five-day period. The assembled peptide was also capable of inducing hydroxyapatite nucleation de novo. The results suggest that self-assembling peptides may be useful in the modulation of mineral behavior during in situ dental tissue engineering.

Potential inhibition of demineralization in vitro by fluoride-releasing sealants

The incorporation of fluoride into sealants has been viewed as a viable way to prevent pit-and-fissure caries by potential inhibition of demineralization through the release of fluoride to enamel. The authors conducted a study to examine the effect of a recently introduced fluoride-releasing sealant (ProSeal, Reliance Orthodontic Products, Itasca, Ill.) on enamel demineralization in an in vitro artificial caries system. The authors randomly assigned 45 extracted human third molars to three treatment groups receiving either conventional sealant without fluoride (Group 1), fluoride-releasing sealant (Group 2) or glass ionomer sealant with high fluoride release (Group 3). They placed cavity preparations on the buccal surfaces of the molars and filled them with the assigned material. They placed acid-resistant varnish on the specimens' enamel surfaces to within 1 millimeter of the sealant, leaving a 1-mm rim of sound enamel available for in vitro enamel caries formation. They thermocycled the teeth (500 cycles) in artificial saliva. They subjected the teeth to an in vitro artificial caries challenge for six weeks to produce caries-like lesions in enamel adjacent to the sealant materials. The authors took longitudinal sections from each tooth, immersed them in water and examined them via polarized light microscopy to determine wall lesion frequencies. The mean (+/- standard deviation) lesion depths were 232 +/- 17 micrometers for Group 1, 144 +/- 21 mum for Group 2 and 128 +/- 15 mum for Group 3. The wall lesion frequency was 12 percent for Group 1 and 7 percent for both Groups 2 and 3. There was a significant difference (P < .05) among the fluoride-releasing materials versus the nonfluoride-releasing material. This study indicates that the new fluoride-releasing sealant substantially reduces the amount of enamel demineralization adjacent to the material. ProSeal provided increased demineralization inhibition compared with a conventional sealant containing no fluoride, but less than that shown by a glass ionomer sealant. ProSeal's physical properties and cariostatic effects may allow for applications beyond traditional sealant use.