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PubMed2000

In vitro antimicrobial activity of propolis and Arnica montana against oral pathogens.

Arnica and propolis have been used for thousands of years in folk medicine for several purposes. They possess several biological activities such as anti-inflammatory, antifungal, antiviral and tissue regenerative, among others. Although the antibacterial activity of propolis has already been demonstrated, very few studies have been done on bacteria of clinical relevance in dentistry. Also, the antimicrobial activity of Arnica has not been extensively investigated. Therefore the aim here was to evaluate in vitro the antimicrobial activity, inhibition of adherence of mutans streptococci and inhibition of formation of water-insoluble glucan by Arnica and propolis extracts. Arnica montana (10%, w/v) and propolis (10%, w/v) extracts from Minas Gerais State were compared with controls. Fifteen microorganisms were used as follows: Candida albicans--NTCC 3736, F72; Staphylococcus aureus--ATCC 25923; Enterococcus faecalis--ATCC 29212; Streptococcus sobrinus 6715; Strep. sanguis--ATCC 10556; Strep. cricetus--HS-6; Strep. mutans--Ingbritt 1600; Strep. mutans--OMZ 175; Actinomyces naeslundii--ATCC 12104, W 1053; Act. viscosus OMZ 105; Porphyromonas gingivalis; Porph. endodontalis and Prevotella denticola (the last three were clinical isolates). Antimicrobial activity was determined by the agar diffusion method and the zones of growth inhibition were measured. To assess cell adherence to a glass surface, the organisms were grown for 18 h at 37 degrees C in test-tubes at a 30 degree angle. To assay water-insoluble glucan formation, a mixture of crude glucosyltransferase and 0.125 M sucrose was incubated for 18 h at 37 degrees C in test-tubes at a 30 degree angle. Arnica and propolis extracts (20 microl) were added to these tubes to evaluate the % of inhibition of cell adherence and water-insoluble glucan formation. The propolis extract significantly inhibited all the microorganisms tested (p < 0.05), showing the largest inhibitory zone for Actinomyces spp. The Arnica extract did not demonstrate significant antimicrobial activity. Cell adherence and water-insoluble glucan formation were almost completely inhibited by the propolis extract at a final concentration of 400 microg/ml and 500 microg/ml, respectively. The Arnica extract showed slight inhibition of the adherence of the growing cells (19% for Strep. mutans and 15% for Strep. sobrinus) and of water-insoluble glucan formation (29%) at these same concentrations. Thus, the propolis extract showed in vitro antibacterial activity, inhibition of cell adherence and inhibition of water-insoluble glucan formation, while the Arnica extract was only slightly active in those three conditions.

Actinomyces↗

PubMed1997

The phosphoenolpyruvate:sugar phosphotransferase system of oral streptococci and its role in the control of sugar metabolism.

Oral streptococci are sugar-fermentative bacteria comprising at least 19 distinct species and are a significant proportion of the normal microbial population of the mouth and upper respiratory tract of humans. These streptococci transport several sugars by the phosphoenolpyruvate:sugar phosphotransferase system (PTS) which concomitantly catalyzes the phosphorylation and translocation of mono- and disaccharides via a chain of enzymic reactions that transfer a phosphate group from phosphoenolpyruvate to the incoming sugar. A number of PTS components, including HPr, Enzyme I and some Enzymes II, have been studied at the biochemical and/or genetical level in Streptococcus salivarius, Streptococcus mutans and Streptococcus sobrinus. Moreover, compelling evidence indicates that the oral streptococcal PTS is involved in the regulation of sugar metabolism. Results are accumulating suggesting that a protein called IIABMan, as well as the phosphocarrier protein HPr, are key regulatory components that allow these bacteria to select rapidly metabolizable sugars, such as glucose or fructose, over less readily utilizable carbohydrates. Circumstantial evidence suggests that the molecular mechanisms by which oral streptococcal PTS exert their regulatory functions differ from mechanisms in other Gram-negative or Gram-positive bacteria.

Bacterial Proteins↗

PubMedDate not supplied

Site-specific oral colonization of mutans streptococci detected by arbitrarily primed PCR fingerprinting.

The clonal diversity and colonization pattern of mutans streptococci within an individual were studied by using a total of 50 tooth site samples obtained from seven 3- to 7-year-old children. Five children contributed saliva samples. From the 7 subjects, 90, 74, 81, 53, 83, 103, and 114 isolates (a total of 598) representing Streptococcus mutans and Streptococcus sobrinus were included for arbitrarily primed polymerase chain reaction (AP-PCR) fingerprinting. The children harbored one to four different AP-PCR types of mutans streptococci. In 3 of the 7 children, the distribution of AP-PCR types of S. mutans differed depending on tooth site (chi-square test for heterogeneity, p<0.001), suggesting that mutans streptococcal clones may selectively colonize specific hard-tissue sites.

Chi-Square Distribution↗

PubMed1997

Influence of fluoride co-cystallized with sugar on caries development in desalivated rats.

Previous studies have shown that fluoride (F) administered concomitantly with sucrose in drinking water, in the diet, or alone is an effective cariostatic agent. The purpose of the present study was to determine the minimum concentration of F co-crystallized with sugar that may be used to prevent dental caries in rats subjected to a severe cariogenic challenge. Desalivated Sprague-Dawley rats, infected with Streptococcus sobrinus, were placed in a König-Höfer programmed feeder. The rats received 17 meals daily for 21 days as follows: group (1) sucrose and sterile distilled water (s.d.w.); (2) sucrose and 1 part/10(6) F-water; (3) 1 part/10(6) F-sucrose and s.d.w.; (4) 4 parts/10(6) F-sucrose and s.d.w.; (5) 8 parts/10(6) F-sucrose and s.d.w.; (6) sucrose and 10 parts/10(6) F-water. Essential nutrition was administered by gavage. At death, blood was collected from each animal and one-half of the lower jaw was sonicated in 0.9% saline solution for microbial assessment and F analysis. Keyes smooth-surface and sulcal caries scores were significantly lower in the groups that received 10 parts/10(6) F-water and 8 parts/10(6) F-sucrose than in all other groups. The F concentration in the jaw suspension and plasma were significantly higher in the 10 parts/10(6) F-water and the 8 parts/10(6) F-sucrose groups than in all other groups. The total cultivable flora and Strep. sobrinus populations were lowest in F groups but this did not reach statistical significance. It is concluded that 8 parts/10(6) F co-crystallized with sucrose reduces the cariogenic potential of sugar as effectively as 10 parts/10(6) F in water, that is as little as 1 part/10(6) in sucrose has a significant effect, and that this cariostatic action is related to the amount of F in the oral environment.

Animals↗

PubMed2006

Influence of a fluoride mouthrinse on mutans streptococci in schoolchildren.

This study aimed to determine whether the long-term use of a fluoride mouthrinse affects the salivary levels of mutans streptococci. Two hundred and fifteen schoolchildren (aged 9-10 years) participated. One hundred and forty-nine of these children had used a fluoride mouthrinse since 5 years of age at nursery school, and the remaining 66 children had not. DFT (decayed and filled teeth) was recorded, and the salivary levels of Streptococcus mutans and Streptococcus sobrinus were measured using mitis salivarius bacitracin agar. The group that had used a fluoride mouthrinse had a significantly lower prevalence of both S. mutans and S. sobrinus (p = 0.038) and a significantly lower DFT score (p < 0.001) than the other group. Using logistic regression analysis including caries experience at baseline as a dependent variable, the odds ratio of carrying S. mutans alone was 8.0 (p = 0.066) and that of carrying both S. mutans and S. sobrinus was 16.5 (p = 0.022) in the group that had not used the fluoride mouthrinse. Children carrying both S. mutans and S. sobrinus had a higher caries incidence in 1 year than the others, with odds ratios of 5.73 (p = 0.067) in the group with a fluoride mouthrinse and 3.47 (p = 0.035) in the group without it. These results show that the long-term use of a fluoride mouthrinse is associated with reduced salivary levels of mutans streptococci and this bacterial reduction may partly contribute to the suppression of dental caries in children using a long-term fluoride mouthrinse.

Cariostatic Agents↗

PubMed1990

[Detection of mutans streptococci by latex agglutination test and its application as a caries-activity test].

The number of mutans streptococci in saliva and dental plaque has been reported to correlate with the incidence of dental caries. This report describes a simple and rapid diagnostic method for the detection of mutans streptococci in dental plaque using latex agglutination (LA) test. Latex particles (0.876 microns, diameter) were sensitized with partially purified antibodies against whole cells of Streptococcus mutans MT8148 (c), MT703R (e) and OMZ175 (f) and Streptococcus sobrinus B13 (d) and 6715 (g). Whole cells of mutans streptococci or dental plaque was extracted with a mixture of 8M sodium nitrite (5 microliters) and 2M acetic acid (5 microliters) for five minutes and neutralized with 2M sodium hydroxide (10 microliters), and the extract and the sensitized latex suspension (20 microliters) were mixed and the grade of agglutination reaction was read macroscopically after ten minutes standing at room temperature. The LA tests could detect 1.0 10ng of purified serotype antigen and 10(5)-10(6) CFU of live cells of mutans streptococci and specifically distinguish not only the mutans streptococci from the other streptococci but also S. mutans from S. sobrinus. However, cross-reactions were still observed among the serotypes c, e and f of S. mutans or between the serotypes d and g of S. sobrinus. Plaque samples were collected from 168 children (2 to 12 years of age) and the 0.1 mg (wet weight) were applied to the LA tests. At the same time, the total number of mutans streptococci in plaque and the serotypes of each isolate were determined. The results of LA reaction correlated significantly with the number of mutans streptococci in plaque (chi-square analysis; p less than 0.0001). The LA tests discriminated between S. mutans and S. sobrinus in dental plaque. It was found that the latex particles sensitized with anti-serotype c and/or e S. mutans antibodies were most effective in demonstrating mutans streptococci, and they were used in the following studies. The results of LA reaction correlated significantly with both the number of decayed tooth surfaces and the number of decayed and filled tooth surfaces. These results suggest that the LA test in mutans streptococci can be useful for the detection of mutans streptococci in dental plaque and also as a caries-activity test at dental clinics.

Chi-Square Distribution↗

PubMed2006

In vitro anti-cariogenic activity of dichloromethane fraction from Rheum undulatum L. root.

This study aimed to evaluate in vitro effects of Rheum undulatum L. root on the development of dental caries, especially its effects on viability, dental plaque formation, and glycolytic acid production of Streptococcus mutans and Streptococcus sobrinus. Methanol extract of Rheum undulatum L. root and its fractions were prepared and tested. Among the test extract and fractions, dichloromethane fraction (DF) showed the most active antibacterial activity (inhibition zone: 13-17 mm) against S. mutans and S. sobrinus in a disc diffusion method. Minimal inhibitory concentrations (MICs) of DF against these bacteria ranged from 0.25 to 0.5 mg/mL. Furthermore, DF significantly inhibited the caries-inducing factors of these bacteria. At sub-MIC levels, DF inhibited in vitro dental plaque formation by S. mutans and S. sobrinus (IC50= 0.079 and 0.142 mg/mL, respectively), which was caused, in part, by the inhibitory effect on the activity of glucosyltransferases. A significant reduction of glycolytic acid production was found at the concentration as low as 0.032 mg/mL for S. mutans and 0.063 mg/mL for S. sobrinus. The possible bioactive compounds that are inducing in vitro anti-cariogenic activity of DF are unknown. Based on the preliminary phytochemical analysis, the activity of DF may be related to the presence of anthraquinones, cardiac glycosides, coumarines, sterols/terpenes, and phenolics. These results indicate that DF is probably useful for the control of dental plaque formation and subsequent dental caries development.

Anti-Bacterial Agents↗

PubMedDate not supplied

Anticaries effect of a component from shiitake (an edible mushroom).

The caries-inhibiting effect of the extract from shiitake (Lentinus edodes), the most popular edible mushroom in Japan, was studied both in vitro and in vivo. Shiitake extract showed an inhibitory effect on water-insoluble glucan formation from sucrose by crude glucosyltransferases of Streptococcus mutans JC-2 and Streptococcus sobrinus OMZ-176. The firmly adherent plaque in the artificial plaque formation test was strongly inhibited by shiitake extract. The reduction of firmly adherent plaque caused an increase in the incidence of non- and loosely adherent plaque and a decrease in total plaque formation. A significantly lower caries score was observed in specific pathogen-free rats infected with S. mutans JC-2 and fed with a cariogenic diet containing 0.25% shiitake extract as compared with controls fed the cariogenic diet without shiitake extract.

Animals↗

PubMed1994

Dental plaque inhibition by a combination of triclosan and polydimethylsiloxane (silicone oil).

It has previously been shown that a toothpaste which contains triclosan and polydimethylsiloxane (silicone oil) improved gingival health in a test panel with established gingivitis more markedly than usually observed when testing commercial triclosan-containing toothpastes. In the present study the mode of action of combinations of silicone oil and triclosan was investigated. An in vitro experiment showed that triclosan was retained in a thin layer of silicone oil on the inner surfaces of test tubes that had been treated with the combination of triclosan and silicone oil. This triclosan was slowly released and inhibited the growth of bacteria in the test tubes. Silicone oil alone and in combination with triclosan was able to inhibit polysaccharide adhesion to the test tubes in the presence of a strain of Streptococcus sobrinus and a sucrose-containing medium. Corresponding in vivo experiments showed similar results; teeth topically treated with silicone oil and triclosan showed a marked plaque reduction, and those treated with silicone oil alone showed a moderate reduction, as compared with a placebo. The silicone oil plus 0.3% triclosan combination inhibited plaque formation also on proximal surfaces. It appears likely that this combination provides a reservoir of triclosan in the thin layer of silicone oil which binds strongly to teeth because of its low surface tension. Triclosan is then slowly released into saliva because of its low solubility in this fluid.

Adult↗

PubMed2001

A novel approach for purification of recombinant proteins using the dextran-binding domain.

Using the dextran-binding domain (DBD) of a type of glucosyltransferase (GTF) from Streptococcus sobrinus, we have developed a novel method for purifying recombinant proteins. DBD-tagged green and red fluorescent proteins as well as the parent GTF and DBD moiety were adsorbed well to commercially available cross-linked dextran (such as Sephadex beads and Sephacryl beads), and eluted efficiently with water-soluble dextran. The purity of the eluted proteins after this one-step affinity purification was approximately 90% or better. The results suggest that DBD can be used as a powerful carrier for purification of various recombinant proteins.

Acrylic Resins↗

PubMed1996

Effect of copper co-crystallized with sugar on caries development in desalivated rats.

The purpose of the present study was to explore the effects of varying levels of copper incorporated in sucrose by co-crystallization on caries development in the desalivated rat model and to examine the retention of copper in the oral cavity and its influence on protein and carbohydrate composition of plaque. Forty-eight Sprague-Dawley rats were infected by Streptococcus sobrinus 6715 and desalivated when aged 26 days. Four groups were placed in a König-Höfer programmed feeder and received 17 meals daily at hourly intervals, and essential nutrition (NCP No. 2) by gavage twice daily. The control group (1) received meals of plain sucrose, and groups 2, 3, and 4 received sucrose co-crystallized with 300 ppm Cu2+(2), 150 Cu2+(3), and 75 Cu2+(4). After 3 weeks, the animals were killed, and blood was collected for determination of copper. The lower jaw was removed and sonicated in 0.9% saline solution. Microbial assessment and copper, protein and carbohydrate assays were performed for each animal. Smooth-surface and [sulcal] caries scores were: (1) 92.0[41.4]; (2) 6.1[34.2]; (3) 16.7[32.0]; (4) 24.0[36.0]. Copper in sucrose drastically affected the population of S. sobrinus when compared with the control. Also, Cu2+ sucrose groups had significantly more copper per unit of protein and carbohydrate in jaw suspension than the control group. Concentration of copper in the blood was apparently unaffected by any dietary regimen. It is concluded that Cu2+ in a range of concentrations (75-300 ppm Cu2+) when delivered co-crystallized with sucrose is an effective cariostatic agent. The co-crystallization of a cariostatic agent with sucrose may be an effective method to decrease the cariogenic potential of sucrose.

Analysis of Variance↗

PubMed2000

Effects of caffeine and theophylline on the development of dental caries in rats.

The purpose of this study was to evaluate the influence of caffeine and theophylline on the development of dental caries in rats. Six Wistar dams (spf), mutans streptococci free, were obtained, each with six male pups. The dams were infected by Streptococcus sobrinus 6715 and divided into three groups which received during the lactating period: (1) diet 2000; (2) diet 2000 plus caffeine (2 mg/100 g) and (3) diet 2000 plus theophylline (0.57 mg/100 g). After weaning, the pups were infected by S. sobrinus, placed in a König-Höfer programmed feeder machine, and received 17 meals daily at hourly intervals, for five weeks. During this time the pups were fed with the same diet that their dams were. The percentage of S. sobrinus relative to total flora was significantly higher in the theophylline group. The results for slight (Ds) and moderate (Dm) dentine lesions, for smooth-surface and sulcal scores were statistically higher for the theophylline group than the other groups. Salivary assays did not demonstrate significant inorganic alterations in salivary composition. Caffeine and theophylline groups showed the highest ulcer score. It is concluded that caffeine does not affect the cariogenic potential of the diet, however theophylline can increase the development of dental caries, and this effect may be related to organic alterations of salivary composition.

Animals↗

PubMedDate not supplied

Cariogenic [correction of Carcinogenic] potential of a typical cassava flour from the Amazonian region of Brazil.

The purpose of this study was to compare the cariogenic potential of a typical cassava flour (CF) with sucrose and starch, using a severe cariogenic challenge model in rats. Thirty Wistar female pups with their dams (mutans streptococci free) were infected by Streptococcus sobrinus 6715, desalivated when aged 25 days, and placed in a Konig-Hofer programmed feeder at age 26 days. They received 17 meals daily at hourly intervals for 21 days as follows: group (1) powdered plain sucrose and sterile distilled water ad libitum (sdwal); (2) Lf and Sdwal (3) powdered starch and sdwal. Essential nutrition was administered by gavage. Data were analyzed by ANOVA. The percentage of S. Sobrinus related to the total flora and the number of this microorganism were higher in the sucrose and CF groups than starch group. Smooth-surface and [sulcal] caries scores for the groups were: (1) 105.5 [48.0]; (2) 34.1 [39.2]; (3) 10.2 [18.1]. All the groups were statistically significantly different from each other (p < 0.01), although the result for sulcal score for CF was very close to the sucrose group. It is concluded that cassava flour, the main source of carbohydrate for the Amazonian population of Brazil, has moderate cariogenic potential.

Animals↗

PubMed2000

Influence of (S)-ketoprofen and fluoride on caries in rats.

OBJECTIVE: Non-steroidal anti-inflammatory agents (e.g., ketoprofen) used topically appear to be effective in reducing bone loss in the ligature model of periodontitis. Ketoprofen, in common with some food preservatives, e.g., benzoate and sorbate, is a weak acid. Fluoride, too, may behave as a weak acid and, similar to the other agents, may exert antibacterial effects. The purpose of the present study was to determine whether a combination of (S)-ketoprofen, an enantiomer of ketoprofen, alone or in combination with fluoride, would suppress Streptococcus sobrinus populations and reduce the incidence of dental caries in rats. MATERIALS AND METHODS: Toothpastes containing ketoprofen and/or monofluorophosphate were applied to the teeth of six groups of 20 rats twice daily for 5 weeks. RESULTS: Fewest S. sobrinus were found in the group treated with a paste containing 3% (S)-ketoprofen + 0.1% F. This group also displayed the lowest incidence of smooth surface caries of all groups. Severity of sulcal surface caries was also lowest in this group. CONCLUSIONS: Results from this study show that the (S) enantiomer of ketoprofen enhances the caries protective effect of fluoride. It is conceivable that this combination could be effective in combating the two most common maladies of the mouth; periodontal disease and dental caries.

Analysis of Variance↗

PubMed2006

Modeling shifts in microbial populations associated with health or disease.

Stable microbial communities associated with health can be disrupted by altered environmental conditions. Periodontal diseases are associated with changes in the resident oral microflora. For example, as gingivitis develops, a key change in the microbial composition of dental plaque is the ascendancy of Actinomyces spp. and gram-negative rods at the expense of Streptococcus spp. We describe the use of an in vitro model to replicate this population shift, first with a dual-species model (Actinomyces naeslundii and Streptococcus sobrinus) and then using a microcosm model of dental plaque. The population shift was induced by environmental changes associated with gingivitis, first by the addition of artificial gingival crevicular fluid and then by a switch to a microaerophilic atmosphere. In addition to the observed population shifts, confocal laser scanning microscopy also revealed structural changes and differences in the distribution of viable and nonviable bacteria associated with the change in environmental conditions. This model provides an appropriate system for the further understanding of microbial population shifts associated with gingivitis and for the testing of, for example, antimicrobial agents.

Actinomyces↗

PubMed1992

Noncariogenicity of erythritol as a substrate.

Erythritol is a sugar alcohol produced by Aureobasidium sp. from glucose. It is 75-80% as sweet as sucrose and is also nonhygroscopic. The aim of this study was to evaluate this sugar substitute from a cariological point of view. Erythritol was neither utilized as a substrate for the lactic acid production nor for plaque formation of mutans streptococci (serotypes a-h) and certain oral microorganisms. It was not utilized for water-insoluble glucan synthesis or cellular adherence by glucosyltransferase from Streptococcus mutans PS-14 (c) and Streptococcus sobrinus 6715 (g). Finally, a significantly lower caries score (3.1 +/- 0.5; mean +/- SEM) was observed in specific pathogen-free rats infected with S. sobrinus 6715 and fed with a diet containing 26% erythritol, as compared to control rats fed with a diet containing 26% sucrose (60.5 +/- 2.0). Also, rats provided a diet containing 56% erythritol chocolate (23.8% erythritol) and challenged with S. mutans PS-14 exhibited a significantly lower caries score (6.7 +/- 0.8) compared to the sucrose chocolate group (82.8 +/- 2.8). The main conclusion from this study is therefore that erythritol is a promising sugar substitute from a cariological point of view.

Actinomyces↗

PubMed1995

Characterization of glucosyltransferaseB, GtfC, and GtfD in solution and on the surface of hydroxyapatite.

GlucosyltransferaseB, GtfC, and GtfD were purified by hydroxyapatite column chromatography, followed by ultrafiltration from the culture supernatant fluids of three Streptococcus milleri constructs (gift from Dr. H.K. Kuramitsu) which harbored individual gtf genes of Streptococcus mutans GS5. GtfB, GtfC, and GtfD were enzymatically active both in solution and in an experimental pellicle (HA-CWS-Gtf) formed by adsorbing Gtf onto the surface of clarified human whole saliva (CWS)-coated hydroxyapatite (HA). The Km values for sucrose for all three enzymes were lower when the enzyme was adsorbed to a surface, compared with when it was in solution. In solution phase assays, and in the absence of primer dextran, glucan production was enhanced 75% when both GtfB and GtfD were present in the reaction mixture, compared with the sum of the individual enzyme activities (p < 0.005). This enhancement did not occur when GtfC was additionally present, or when the GtfB+GtfD enzyme pair was adsorbed onto HA-CWS. In additional experiments, glucan formed by GtfB or GtfC, but not by GtfD, on a HA-CWS-Gtf surface increased adherence of Streptococcus mutans GS5 and Streptococcus sobrinus 6715 by seven- to nine-fold compared with adherence when no glucan was present on the pellicle surface (p < 0.001). Further, treatment of the HA-CWS-GtfB-glucan or HA-CWS-GtfC-glucan pellicle with alpha-1,6 dextranase significantly reduced adherence of both streptococcal strains (p < 0.001). These results show that GtfB, GtfC, and GtfD are enzymatically active in an adsorbed state and that the nature of their product glucan can influence the adherence of cariogenic oral streptococci to an experimental pellicle.

Adsorption↗

PubMed1992

The effect of phenytoin medication on dentin apposition, root length, and caries progression in rat molars.

To determine the effect of phenytoin (PHT) on dentin apposition and the rate of caries progression in dentin, 40 Wistar rats were treated daily for 35 days with intraperitoneal injections of 5,5-diphenylhydantoin. Twenty-eight controls received saline with pH adjusted to that of PHT. One PHT and one control group were fed a 43% sucrose diet, and the others a non-cariogenic pellet diet. Streptococcus sobrinus was inoculated to induce caries. Schiff staining was used to determine caries. The areas of dentin apposition and dentinal caries lesions were quantified after tetracycline staining. The root lengths were measured. PHT reduced slightly the dentin apposition and activated significantly the progression of the dentinal carious lesions. No difference in caries initiation was found. The high-sucrose diet reduced dentin apposition and increased the rate of progression of existing caries greatly. Our study suggests that both the high-sucrose diet and PHT have an effect on secreting odontoblasts, which can be seen as an alteration in dentin apposition and caries progression rates in dentin.

Animals↗