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Pre-clinical evaluation of the anticaries effect of an experimental Malva sylvestris extract mouthwash using a cariogenic model in situ.

OBJECTIVE: The aim of this study was to evaluate the antimicrobial and anticariogenic potential of Malva sylvestris extract on enamel and dentin in situ. METHODS: A double-blind crossover in situ study was conducted with 12 participants wearing palatal appliances containing two bovine enamel and two dentin specimens per 3 phases, a total of 72 enamel and dentin specimens. Biofilm formation and daily sucrose exposure were allowed. Treatments were applied twice daily in three phases: Malva sylvestris (2.5%, MS); fluoride (225 ppm, F); and placebo (P). After seven days, biofilm was collected from the bovine specimens for analysis of Lactobacillus spp. and mutans streptococci by Colony Forming Unit counts (CFU log₁₀/mL). Dental demineralization of the bovine specimens was assessed by transverse microradiography (TMR). RESULTS: MS did not reduce Lactobacillus spp. counts (CFU log₁₀/mL: enamel 6.63±0.81; dentin 6.68±0.92) compared to P (6.63±0.70; 6.62±0.51). F also did not differ (6.29±0.75; 6.32±0.41; ANOVA/Tukey, p>0.38). Mutans streptococci data were inconclusive. In enamel, both MS (2320.8±768.2 %vol·µm; 101.6±27.0 µm) and F (1777.3±733.3 %vol·µm; 95.8±23.5 µm) significantly reduced integrated mineral loss and lesion depth compared to P (3517.2±1119.9 %vol·µm; 138.6±19.5 µm; ANOVA/Tukey, p≤0.0003). In dentin, MS significantly reduced integrated mineral loss (322.5 [250-580] %vol·µm) and lesion depth (30.1 [15-42.2] µm) compared to P (880 [580-1705]; 58.3 [32.2-88.6] µm; Kruskal-Wallis/Dunn, p≤0.001), while F (587.5 [305-720]; 25.2 [16.5-38.8] µm) did not differ significantly (p>0.05). CONCLUSIONS: Malva sylvestris extract had no antimicrobial effect on Lactobacillus spp. counts, but significantly reduced enamel and dentin demineralization, showing anticaries effect comparable to fluoride. CLINICAL RELEVANCE: Malva sylvestris has demonstrated promising biological activity. This study investigates the antimicrobial efficacy of Malva sylvestris against cariogenic microorganisms in situ. Our findings provide relevant evidence that M. sylvestris exert significant anticaries effects using an in situ model.

Biofilms↗

Ultrastructure of smear layer-covered intraradicular dentin after irrigation with BioPure MTAD.

The structure of mechanically instrumented intraradicular dentin after irrigation with NaOCl as the initial rinse and BioPure MTAD as the final rinse were examined from the coronal, middle, and apical parts of root canal walls using transmission electron microscopy. Sterile distilled water and EDTA as final rinses were employed as the respective positive and negative controls under the same experimental conditions. There were 2 to 5 mum thick smear layers produced on mechanically instrumented root canal walls that were completely removed by EDTA and BioPure MTAD under agitation. Both irrigants created a zone of demineralized collagen matrices in eroded dentin and around the dentinal tubules, with the mildly acidic BioPure MTAD being more aggressive than EDTA. These demineralized dentin zones create the opportunity for dentin hybridization by infiltration of hydrophilic adhesives/sealers. However, the potential consequences of compaction of hydrophobic sealers against air-dried, collapsed collagen matrices, and hydrolytic degradation of incompletely infiltrated matrices remain unresolved.

Bicuspid↗

Effect of a calcium carbonate-based dentifrice on enamel demineralization in situ.

Since the effect of calcium carbonate (CaCO(3)) based dentifrice on enamel demineralization is not clearly established, it was evaluated using the IEDT model described by Zero's group in 1992. This study had a crossover design and 10 volunteers were submitted to 3 treatment groups: a negative control, brushing without dentifrice; an active control, brushing with silica-based dentifrice (SiO(2)/MFP group), and the experimental group, brushing with CaCO(3)-based dentifrice (CaCO(3)/MFP). Both dentifrices contained 1,500 microgram F/g (w/w) as sodium monofluorophosphate (MFP). Enamel surface microhardness was determined in the dental blocks and the percentage change in relation to baseline was calculated. Fluoride uptake in enamel and its concentration in 'test plaque' were determined. The results showed that the dentifrice containing CaCO(3)/MFP was more effective than SiO(2)/MFP in reducing enamel demineralization (p < 0.05). A higher concentration of fluoride ion was found in 'test plaque' treated with CaCO(3)/MFP than in the negative control (p < 0.05). The results suggest that CaCO(3) abrasive may enhance the effect of fluoride present in dentifrice on dental caries control.

Adult↗

Chemical, morphological and thermal effects of 10.6-microm CO2 laser on the inhibition of enamel demineralization.

Studies have shown that enamel can be modified by pulsed CO2 laser to form a more acid-resistant substrate. This study evaluated the effects of a 10.6-microm CO2 laser on enamel surface morphology and chemical composition as well as monitored intrapulpal temperature changes during irradiation. Human teeth were irradiated with fluences of 1.5-11.5 J/cm2, and pulpal thermal as well as chemical and morphological modifications on enamel were assessed. The teeth were submitted to a pH-cycling model, and the mineral loss was determined by means of cross-sectional microhardness. For all irradiated groups, intrapulpal temperature changes were below 3 degrees C. FT-Raman spectroscopy and scanning electron microscopy indicated that fluences as low as 6.0 J/cm2 were sufficient to induce chemical and morphological changes in enamel. Then, for fluences reaching or exceeding 10.0 J/cm2, laser-induced inhibitory effects on demineralization were observed. It was thus concluded that laser energy density in the range of 10.0 and 11.5 J/cm2 could be applied to dental enamel in order to produce chemical and morphological changes and reduce the acid reactivity of enamel without compromising the pulp vitality.

Analysis of Variance↗

Quantification of root caries using optical coherence tomography and microradiography: a correlational study.

PURPOSE: The use of transverse microradiography (TMR) to quantify the amount of mineral lost during demineralization of tooth tissue has long been established. In the present study, the use of an en-face Optical Coherence Tomography (OCT) technology to detect and quantitatively monitor the mineral changes in root caries was investigated and correlated with TMR. MATERIALS AND METHODS: We used an OCT system, developed initially for retina imaging, and which can collect A-scans, B-scans (longitudinal images) and C-scans (en-face images) to quantitatively assess the development of root caries. The power to the sample was 250 microW, wavelength lambda = 850 nm and the optical source linewidth was 16 microm. RESULTS: Both the transversal and longitudinal images showed the caries lesion as volumes of reduced reflectivity. Quantitative analysis using the A-scan (reflectivity versus depth curve) showed that the tissue reflectivity decreased with increasing demineralization time. A linear correlation (r = 0.957) was observed between the mineral loss measured by TMR and the percentage reflectivity loss in demineralized tissue measured by OCT. CONCLUSION: We concluded that OCT could be used to detect incipient root caries, and that the reflectivity loss in root tissue during demineralization, measured by OCT, could be related to the amount of mineral lost during the demineralization.

Analysis of Variance↗

Effect of timing of administered calcium lactate on the sucrose-induced intraoral demineralization of bovine enamel.

A number of soluble calcium salts are known to reduce the demineralization of enamel in the mouth. The present study was undertaken to examine the effects of rinses containing different concentrations of calcium lactate, and the time of giving the rinses with respect to sucrose challenges. Subjects wore palatal appliances containing blocks of bovine enamel whose surfaces were covered with Streptococcus mutans IB 1600, and rinsed with 10% sucrose for 1 min. Changes in iodide penetrability of the enamel, and the pH and extracellular ion concentrations of the streptococcal plaque were determined. When added to the sucrose rinse, 100 or 150 mM calcium lactate reduced demineralization by about 35%, although the plaque pH was not affected. Plaque calcium was elevated but diffused away rapidly so that concentrations after 45 min were close to control values. Plaque inorganic phosphate and lactate were not affected. Ongoing demineralization appeared to be stopped when 100 mM calcium lactate was given 15 min after the sucrose rinse. When the lactate was given 15 min before the sucrose rinse, demineralization was reduced by only about 25%, consistent with the rapid diffusion of plaque calcium. The combination of (i) pretreatment with calcium lactate and (ii) admixture of calcium lactate with sucrose was most effective. Demineralization was reduced about 55% with 100 mM calcium lactate under these conditions, and protective effects were seen with as little as 25 mM. In summary, the findings demonstrate the enamel-protective effect of relatively low concentrations of calcium lactate, and point to the need to sustain a high plaque calcium during periods of maximum acidogenicity.

Adult↗

Dental caries process.

The boundaries of caries diagnosis and caries intervention are changing. Dentists currently use visual, tactical, and radiographic information to detect relatively advanced changes in the dental hard tissues. The clinical management of dental caries has been primarily directed at the treatment of the consequences of the disease process by placing restorations and not at curing the disease. Using emerging technology, dentists will be able to detect incipient dental caries (demineralization) at an earlier stage than the clinically visible white spot. Dental caries is a dynamic process, which in its early stages is reversible and even in its more advanced stages can be arrested. The widespread use of fluoride has dramatically reduced the prevalence of dental caries and the rate of the progression of carious lesions. These changes permit dentists to adopt more conservative management strategies directed at the prevention and cure of dental caries.

Cariostatic Agents↗

Effects of a supersaturated pulpal fluid on the formation of caries-like lesions on the roots of human teeth.

Root surfaces were exposed to demineralizing buffer in a continuous-flow system, with or without simultaneous perfusion of the pulp chamber with a supersaturated surrogate dentine fluid (SDF). Experimental lesions formed with perfusion of the pulp were significantly less deep than control lesions formed without perfusion, but the application of hydrostatic pressure to the SDF (15 or 30 cm water) did not produce significantly greater reductions. The lesion depth reduction was attributed to reduced undersaturation at the advancing lesion front through interdiffusion of the demineralizing buffer and the SDF. Translucent bands, resembling sclerotic dentine, were observed beneath 9 of 24 experimental lesions and 5 of 24 control lesions. Scanning electron microscopy showed that, in most of the bands beneath experimental lesions, dentinal tubules were either partly occluded by granular mineral deposits or were reduced in diameter. Thus, in vivo, dentine fluid may modify the rate of lesion progression and promote sclerosis in the underlying dentine.

Acetates↗

The effect of a low fluoride containing toothpaste on the development of dental caries and microbial composition using a caries generating model device in vivo.

The purpose of the study was to evaluate the effect of daily use of a low fluoride containing toothpaste (250 ppm F) on the uptake of fluoride and development of enamel lesions as well as the prevalence of lactobacilli and mutans streptococci in dental plaque compared to the use of placebo toothpaste. 16 children were selected with homologous premolar teeth. The teeth were cemented with orthodontic bands ad modum Ogaard for plaque accumulation and enamel lesion development. The plaque accumulated during 4 weeks was collected and analysed for lactobacilli and mutans streptococci. The teeth were further analysed by secondary ion mass spectrometry (SIMS), determining the concentration profiles of fluoride and other elements in the outermost enamel and in the lesion. The results show that although significant amounts of fluoride were taken up in the surface enamel from the fluoride toothpaste, the extent of the lesions was not influenced compared to teeth brushed with a non F-toothpaste. Neither were microbiological differences in the dental plaque found between the groups. An interesting observation was that early demineralization of enamel took place without detectable levels of mutans streptococci in the overlaying dental plaque. The conclusion is that fluoride taken up in enamel from F-toothpaste has no significant influence on enamel lesion development if a cariogenic dental plaque with high levels of acid producing microorganisms is continuously attached to the enamel surface.

Adolescent↗

Validation of the intraoral delta Ip system and use of the system to test the efficacy of Mentadent dentifrice.

Ongoing efforts within the oral care industry to improve dentifrices by the addition of anti-tartar compounds, bicarbonate and other agents, have necessitated the continuing evaluation of new products in order to assure their anti-caries effectiveness. However, clinical evaluation of new products is time consuming, prohibitively expensive and ethically questionable. Recently, a number of intraoral models have been developed that appear to be well-suited to evaluate new products. In the present study, an intraoral system was used that monitors short-term demineralization of surface enamel. Subjects wore appliances that contained Streptococcus mutans-coated blocks of bovine enamel and rinsed first with 15 ml of a 20% slurry of dentifrice and, after 30 minutes, with a 10% sucrose solution. Iodide penetrability (Ip) was determined before and after each intraoral exposure, and the difference, or delta Ip, was taken as the measure of demineralization. The model system responded linearly to increasing concentrations of fluoride in a series of standard dentifrice preparations. Mentadent, a fluoride toothpaste with baking soda and peroxide, was shown to reduce enamel demineralization and to provide protection comparable to a clinically tested dentifrice (Crest Regular). These results established the validity of the delta Ip method, and demonstrated the efficacy of Mentadent according to the criteria established at the ADA Consensus Conference on Intraoral Models in 1990.

Adult↗

Scanning and contact microradiographic study of the effect of degree of saturation on the rate of enamel demineralization.

The effect of degree of saturation with respect to hydroxyapatite (DSHA) on enamel section demineralization was studied by contact and scanning microradiography (CMR and SMR). Two aspects were studied: (1) the effect of different values of DSHA at constant pH, and (2) the effect of constant DSHA with variable pH. In the pH range (2.5-4.5) and time scale (0-120 h) studied, the DSHA was more important in determining the rate of enamel lesion progression than was the pH.

Adult↗

Effect of luting cement on dental biofilm composition and secondary caries around metallic restorations in situ.

Since the importance of luting cement on secondary caries in enamel and dentin is unknown, an in situ crossover study was conducted in three phases over 21 days using a fluoride-containing toothpaste. One hundred and twenty-six metallic restorations were cemented into the dentinoenamel junction of slabs of human teeth with zinc phosphate (ZP), resin-modified glass ionomer (GI) or resinous cement (RC). The slabs were inserted onto flanges of the removable partial acrylic dentures of 14 volunteers and covered with gauze to enhance dental plaque accumulation. The volunteers used fluoride toothpaste (1.100 microg F/g, w/w). After 21 days, the biofilm that formed on the slabs was collected for biochemical and microbiological analyses, and the demineralization in enamel-dentin around the restorations was evaluated. The fluoride concentration of biofilm in the GI group was higher (p<0.05) than the ZP and RC groups. Also, the concentration of Zinc in biofilm formed on the slabs cemented with ZP was higher (p<0.05) than the other groups. However, the effect of the luting material on enamel or dentin demineralization was not statistically significant (p>0.05). The data suggest that when fluoride toothpaste is used, the anticariogenic property of the luting cement may not be relevant to the reduction of secondary caries.

Adult↗

Effects of chlorhexidine on the bacterial colonization and degradation of dentin and completely demineralized dentin in situ.

The effects of 0.2% chlorhexidine on selected plaque microorganisms were studied in an intraoral dentin caries model. In 8 individuals wearing partial dentures, sound and completely demineralized dentin specimens were placed consecutively in 2 periods of 4 weeks, respectively. Throughout the experimental period, the specimens were treated 2 x daily with 0.2% chlorhexidine; control specimens were treated with water. Plaque accumulation on the specimens was left undisturbed. No protection against demineralization of the dentin or degradation of the dentin collagen by the chlorhexidine treatment was observed. The chlorhexidine treatment did not result in a reduction of the total cultivable flora when compared with the control specimens. A significant reduction of mutans streptococci and total streptococci recovered from completely demineralized dentin treated with chlorhexidine was observed, but the proportions of Actinomyces and lactobacilli were not affected significantly. It is speculated that areas of exposed roots, which are difficult to reach by oral hygiene measurements, such as approximal surfaces, will not be protected by a 0.2% chlorhexidine mouthrinse against the caries process.

Actinomyces↗

Inductive effect of native dentin on the dentinogenic potential of adult dog teeth.

Autogenous dentinal matrix was exposed to the pulp cells of adult dogs in order to determine whether the mature pulp cells possess the ability to differentiate into odontoblast-like cells as a direct response to known inductive influences. The pulps of molars, premolars, and canines of three dogs (2 to 4 yr old) were mechanically exposed through buccal class V cavities. Pieces of demineralized or native dentin and predentin were implanted in the pulp sites for periods of 2 to 6 wk. The reactions were analyzed by light microscopy. Induction of dentin formation was observed only after native dentin implantation; either as early response to exposure of predentinal surfaces or around mineralized dentin after 3 postoperative wk. Encapsulation by fibrous connective tissue or matrix degradation was seen around demineralized dentin implants. A characteristic enhancement of circumferential pulpal dentin deposition around the implantation site was demonstrated after native dentin exposure to light and scanning electron microscopic examination. These data indicate that specific inductive influences given by the native but not the acid-conditioned dentin, when it is exposed to the pulp environment of adult teeth, are able to direct differentiation of odontoblast-like cells and to enhance the biosynthetic activity of primary odontoblasts.

Animals↗

Influence of enamel thickness on quantification of mineral loss in enamel using laser-induced fluorescence.

The objective of this study was to evaluate the effect of enamel thickness on the quantification of white-spot lesions with laser fluorescence. One hundred and twenty 3-mm-diameter human ground and polished enamel specimens were used. Specimens were illuminated by a 488-nm argon laser, images were acquired through a 540-nm high-pass filter and stored on the Quantitative Light-Induced Fluorescence (QLF) program. Specimens were divided into two groups and demineralized for 48 or 96 h and QLF images were again acquired. Fluorescence radiance (FR) of both sound (FR(s)) and demineralized (FR(d)) enamel was determined using the QLF program. Change in fluorescence radiance (Delta FR, %) was determined as follows: Delta FR = FR(d)/FR(s) x 100. One thin section was obtained for transverse microradiography analysis. Enamel and dentin thickness of the thin sections were measured. There was a good correlation between QLF parameters and enamel thickness. It can be concluded that the FR observed for similar lesions depends on the actual enamel thickness.

Dental Enamel↗

Relationships between a clinical-visual scoring system and two histological techniques: a laboratory study on occlusal and approximal carious lesions.

One aim of the present laboratory study was to determine whether a visual scoring system (ERK) developed for occlusal caries could be applied to approximal lesions. A new histological technique (autofluorescence, AF) recognises dentine that is soft and would be removed with an excavator during operative treatment. A second aim was to investigate the relationship between the visual scoring system (ERK) and AF of dentine both occlusally and approximally. The sample comprised 93 extracted teeth chosen to represent the range of visual scores on approximal and occlusal surfaces. After sectioning through the investigation site, the cut faces were examined in a stereomicroscope and the depth of demineralization was scored. Autofluorescence was viewed with a confocal laser scanning microscope. Results showed reasonable correlation between the visual scores and the stereomicroscope histological evaluations for occlusal surfaces and non-cavitated approximal surfaces. However, cavitated approximal surface lesions were less advanced histologically than cavitated occlusal carious lesions. The AF technique indicated that several lesions with intact surfaces would have had soft, excavatable dentine, whereas several with microcavities would not.

Dental Caries↗

The activation and function of host matrix metalloproteinases in dentin matrix breakdown in caries lesions.

Matrix metalloproteinases (MMPs) are a family of enzymes which, in concert, are capable of degrading collagen. We investigated whether human MMPs could participate in the degradation of dentin organic matrix after demineralization. We performed Western blot analyses using MMP-specific antibodies to identify MMPs in human dental caries lesions. Enzymography and functional activity assays, with 125I-labeled gelatin as substrate or quantitating the degradation of type I collagen, were used to determine the activity of purified and salivary gelatinolytic (MMP-2 and MMP-9) and collagenolytic (MMP-8) enzymes with and without acid-activation in pHs relevant to caries. Respective analyses were done with caries-related bacteria. We performed electron microscope analyses to assess the degradative activity of sterilized salivary host MMPs on demineralized human dentin. Human MMP-2, MMP-8, and MMP-9 were identified in demineralized dentinal lesions. The latent purified forms of these enzymes were activated at low pH (4.5), followed by neutralization, mimicking the conditions during caries progression. Incubation of human saliva at low pH followed by neutralization resulted in a four-fold increase in the gelatinolytic activity. No gelatinolytic or collagenolytic activity was observed in bacterial samples. The activated enzymes in saliva degraded demineralized dentin organic matrix in vitro. These results demonstrate the pH-dependent activation mechanism of MMPs, which may have a distinct role in different physiological and pathological conditions. They further demonstrate that host MMPs, activated by bacterial acids, have a crucial role in the destruction of dentin by caries.

Bacterial Proteins↗

Load dependency and reliability of microhardness measurements on acid-etched enamel surfaces.

The load dependency of hardness measurements was investigated on sound and demineralized enamel samples. For all samples investigated, the Knoop hardness number (KHN) varied as a function of the applied load. Higher loads enhanced indenter penetration into the test surface. The variation in KHN values observed in demineralized enamel, both surface-softened and surface-etched, was explained by a decrease in porosity as a function of depth. The reading error for indentations made on surface enamel etched with 37% phosphoric acid gel for 30 s was found to be slightly elevated compared to indentations made on a polished sound enamel surface. It was concluded that the surface microhardness technique can be used for the longitudinal investigation of etched surface enamel.

Acid Etching, Dental↗