The role of grape seed extract in the remineralization of demineralized dentine: micromorphological and physical analyses.

OBJECTIVES: Grape seed extract (GSE) is known to have a positive effect on the demineralization and/or remineralization of artificial root caries lesions. The present study aimed to investigate whether biomodification of caries-like acid-etched demineralized dentine, using proanthocyanidins-rich GSE, would promote its remineralization potential. DESIGNS: Dentine specimens were acid-etched for 30s, then biomodified using proanthocyanidin-based preconditioners (at different concentrations and pH values) for 2min, followed by a 15-day artificial remineralization regimen. They were subsequently subjected to microhardness measurements, micromorphological evaluation and X-ray diffraction analyses. Stability of the preconditioners was also analyzed, spectrophotometrically. RESULTS: A concentration-dependent increase was observed in the microhardness of the specimens that were biomodified using GSE preconditioners, without pH adjustment. Field emission scanning electron microscopy revealed greater mineral deposition on their surfaces, which was further identified mainly as hydroxylapatite. The absorbances of preconditioner dilutions at pH 7.4 and pH 10.0 decreased at the two typical polyphenol bands. CONCLUSIONS: Transient GSE biomodification promoted remineralization on the surface of demineralized dentine, and this process was influenced by the concentration and pH value of the preconditioner. GSE preconditioner at a concentration of 15%, without pH adjustment, presented with the best results, and this may be attributed to its high polyphenolic content.

Biomimetic approach for root caries prevention using a proanthocyanidin-rich agent.

The aim of this study was to evaluate the effects of a proanthocyanidin-rich grape seed extract (GSE) on the in vitro demineralization of root dentine. Root fragments were obtained from sound human teeth. The fragments were randomly assigned to different treatments solutions: GSE, fluoride (F), GSE+F and distilled water (control). Samples were treated daily for 30 min and subjected to a pH cycling artificial caries protocol using demineralization cycles (2.2 mM CaCl(2)×H(2)O, 2.2 mM KH(2)PO(4), 50 mM acetic acid, pH 4.3) for 6 h and remineralization cycles (20 mM HEPES, 2.25 mM CaCl(2)×H(2)O, 1.35 mM KH(2)PO(4), 130 mM KCl, pH 7.0) for 17.5 h. Mineral loss (ΔZ) and lesion depth (LD) were determined after 18 days of treatment/pH cycling, by transverse microradiography. GSE was able to minimize ΔZ and LD compared with the control group (p < 0.0001). The GSE+F and F groups showed the lowest values of ΔZ and LD (p < 0.05), with no statistically significant differences between them (p = 0.554 and p = 0.726, respectively). A biomimetic approach to strengthen root dentine using GSE results in decreased rates of root demineralization and may be used in conjunction with F to prevent root caries.

Reversal of primary root caries lesions after daily intake of milk supplemented with fluoride and probiotic lactobacilli in older adults.

OBJECTIVE: To evaluate the effect of milk supplemented with fluoride and/or probiotic bacteria on primary root caries lesions (PRCL) in older adults. MATERIALS AND METHODS: After informed consent, 160 healthy subjects, 58-84 years of age, with at least two PRCL were recruited and randomly assigned to one of four parallel study groups drinking 200 ml milk once daily for 15 months. Group A consumed standard milk (placebo); Group B ingested milk supplemented with 5 ppm F and probiotic bacteria (Lactobacillus rhamnosus LB21, 10(7) CFU/mL); Group C drank milk with only probiotic bacteria and group D milk contained only fluoride. Primary endpoints were Root Caries Index (RCI) and electric resistance measurements (ECM) carried out by one blinded single examiner. Secondary endpoints were mutans streptococci and lactobacilli counts in saliva and plaque estimated with chair-side tests. Data were compared within and between groups with non-parametric tests. RESULTS: The drop out rate was 38%. At baseline there were no statistical differences between the groups. Significantly higher numbers of RCI reversals were found in groups B, C and D compared with group A (p < 0.05). The mean ECM values increased significantly (p < 0.05) in all groups except for the placebo group A, indicating that remineralization occurred. The effect was most beneficial in the two groups that contained fluoride. No significant alterations were displayed regarding the microbial counts. No severe adverse effects were reported during intervention. CONCLUSION: Daily intake of milk supplemented with fluoride and/or probiotic bacteria may reverse soft and leathery PRCL in older adults.

Relationship between root caries and alveolar bone loss in the first wet-rice agriculturalists of the Yayoi period in Japan.

OBJECTIVE: The people of the Yayoi period (5th C b.c.-3rd C a.d.), who were the first wet-rice agriculturalists in ancient Japan, had carious lesions that were most frequently located on the root surfaces of their teeth. Root surface exposure is a prerequisite for this type of decay, and alveolar bone loss is the main cause of such exposure. Therefore, we identify the factors associated with root caries, and examine the relationship between root caries and alveolar bone loss in the people of the Yayoi period. MATERIALS AND METHODS: This study was performed using 263 sets of ancient skeletal remains that are believed to be from the Yayoi period and that were excavated at 49 archaeological sites in western Japan. Using 5010 teeth found among the remains, we analysed the relationship between the prevalence of root caries and the cemento-enamel junction-alveolar crest (CEJ-AC) distance. RESULTS: The prevalence of root caries and the mean number of teeth with root caries per person were significantly correlated with age, the presence of coronal caries and the mean CEJ-AC distance per person. We also found that as the mean CEJ-AC distance per tooth surface increased, the percentage of the root surface affected by caries increased. Moreover, after excluding the lingual (palatal) side, the mean CEJ-AC distance per surface was significantly greater for those tooth surfaces with root caries. CONCLUSION: We present the first evidence that the occurrence of root caries correlated with the CEJ-AC distance in the Yayoi people of Japan.

In vitro remineralization effects of grape seed extract on artificial root caries.

Grape seed extract (GSE) contains proanthocyanidins (PA), which has been reported to strengthen collagen-based tissues by increasing collagen cross-links. We used an in vitro pH-cycling model to evaluate the effect of GSE on the remineralization of artificial root caries. Sound human teeth fragments obtained from the cervical portion of the root were stored in a demineralization solution for 96 h at 37 degrees C to induce artificial root caries lesions. The fragments were then divided into three treatment groups including: 6.5% GSE, 1,000 ppm fluoride (NaF), and a control (no treatment). The demineralized samples were pH-cycled through treatment solutions, acidic buffer and neutral buffer for 8 days at 6 cycles per day. The samples were subsequently evaluated using a microhardness tester, polarized light microscopy (PLM) and confocal laser scanning microscopy (CLSM). Data were analyzed using ANOVA and Fisher's tests (p<0.05). GSE and fluoride significantly increased the microhardness of the lesions (p<0.05) when compared to a control group. PLM data revealed a significantly thicker mineral precipitation band on the surface layer of the GSE-treated lesions when compared to the other groups (p>0.05), which was confirmed by CLSM. We concluded that grape seed extract positively affects the demineralization and/or remineralization processes of artificial root caries lesions, most likely through a different mechanism than that of fluoride. Grape seed extract may be a promising natural agent for non-invasive root caries therapy.

Micromorphological and micronanalytical characterization of stagnating and progressing root caries lesions.

Root caries is the predominant disease of the tooth tissues in the elderly population and differs in progression and micromorphology from coronal dentin caries. Therefore, different clinical concepts are needed for the treatment of these progressing and arrested lesions. It was the aim of this study to investigate the three-dimensional structure and volume of stagnating and progressing root caries lesions and to determine the mineral composition of the different lesion zones to achieve a better understanding of the natural history of root caries lesion formation. Of 21 extracted human teeth of patients between 42 and 77 years of age with stagnating and progressing root caries lesions serial sections were cut and investigated with polarized light and scanning electron microscopy. From the polarized light micrographs 3D reconstructions were made to determine the volumes of the lesion zones and their relations expressed in a demineralization index (DI). With increasing size of the demineralizing zone the DI increased indicating an increase in the size of the translucent zone. The 3D reconstructions showed distinct differences between stagnating and progressing root caries lesions. In the hypermineralized translucent dentin not all dentin tubules were obliterated by intratubular dentin and within the translucent dentin scattered dead tracts were found. Electron dispersive X-ray (EDX) analysis showed that the intertubular dentin of the translucent dentin was also demineralized to a certain extent. The results indicate distinct morphological differences between progressing and stagnating root caries lesions which may have consequences for treatment strategies.

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

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

Studies of dental root surface caries. 2: The role of cementum in root surface caries.

Artificial caries lesions were produced in roots of teeth using an acetate buffer system, when the layer of cementum was either normal in thickness, excessively thickened by hypercementosis, or had been removed completely. The rates of lesion progression were measured in each case using polarized light microscopy to measure lesion depth. Analysis of calcium (Ca) and phosphorus (P) loss during the demineralizing process was carried out. The removal of cementum was found to significantly increase the initial rate of penetration of the lesion into the root, although this rate progressively reduced to a level consistent with that found in normal roots after seven days of demineralization. The overall depth remained consistently greater than that observed in normal roots, or when lesions were produced entirely within hyperplastic cementum. Chemical analysis also showed removal of cementum resulted in an initial doubling of the Ca and P lost from the root surface. Prior direct exposure of segments of normal roots to the oral environment was found not to significantly alter the rate of artificial lesion progression, in comparison with that in the originally protected segment of the root surface. It was concluded that an intact cementum layer has the intrinsic ability to protect the underlying dentine of exposed tooth roots against acidic demineralization and that prior exposure to the oral environment does not significantly alter this ability.