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Although Hertwig’s epithelial root sheath (HERS) performs an important function in the formation of the tooth root, the developmental mechanisms that control HERS growth and differentiation remain to be thoroughly elucidated. Bone morphogenetic protein 4 (BMP4), which is secreted by mesenchymal cells, acts on the dental epithelium as a regulator of cell differentiation during crown formation. In an effort to determine whether BMP4 specifically regulates the development of HERS in the dental epithelium, we assessed the localizations of BMP4, BMP receptor-IB (BMPR-IB), and BMPR-II during molar root formation in the mouse. HERS cells were shown to express BMPR-IB and BMPR-II. BMP4-positive cells were detected densely in the dental papillae around HERS, thereby suggesting that BMP4 participated in HERS formation. Beads soaked in BMP4, NOGGIN, or phosphate-buffered saline (PBS) were implanted into the pulp cavity under culture conditions, and the length of HERS was evaluated with regard to the proliferating cells. After 12 h, both groups exhibited a similar HERS developmental pattern, with the length and shape of HERS bearing a close resemblance to one another. However, after 48 h, the observed HERS elongation was significantly shorter in the BMP4-treated group. In addition, proliferative cell nuclear antigens were detectable only in the NOGGIN- and PBS-treated groups. These findings demonstrate that mesenchymally expressed BMP4 regulates HERS development by preventing elongation and maintaining cell proliferation. BMP4 may, therefore, prove useful as a root-formation regulatory agent in a variety of tissue-engineering applications.

This study aimed to compare the effectiveness of an experimental root canal irrigant and 17% Ethylene-di-amine tetra acetic acid for removal of the smear layer in the coronal, middle and apical portions of the root canal. Ninety human single rooted maxillary and mandibular teeth were selected for this study. The teeth were randomly divided into two experimental groups and one control group as follows: Group A (Ethanolic extract of Sapindus Mukorossi), Group B (17% EDTA), and Group C (Distilled water). The root canals of all three groups were prepared with stainless steel K-files by means of the standard step-back technique and irrigated with 5.25% sodium hypo chloride. The teeth were decoronated, following the irrigation and divided longitudinally into two-halves and visualized using scanning electron microscope (SEM) for the amount of smear layer present utilizing the three-point score system. The observations were noted both before and after the treatment. Nonparametric tests were applied for the comparison and -value ⩽ 0.05 was considered as statistically significant. It was evident from that smear layer was completely removed in coronal portion of 27 out of 30 teeth in-group A. For middle and apical areas of group A, 24 and 19 teeth showed complete smear layer removal. In-group B it was found that there were 24, 21, and 3 teeth at coronal, middle and apical, areas respectively where smear layer were completely absent. Intra group comparison showed a significant difference (  = 0.002) in smear layer removal was found for group A at coronal, middle and apical thirds. Similarly, a significant difference (  = 0.001) was also found for group B; however heavy smear layer was found among the three parts of the canal for group C. Ethanolic extract of Sapindus Mukorossi have higher effectiveness in removing the smear layer from the root canal in comparison to 17% EDTA.

High-fluoride dentifrice is used to manage root caries, but there is no evidence whether its association with nanohydroxyapatite could provide an additional protection for root caries. Therefore, this study aimed to develop and evaluate the effect of an experimental dentifrice with high fluoride (F ) concentration and nanohydroxyapatite (nano-HA) on root dentin demineralization. After formulation of dentifrices, root dentin specimens were randomly assigned to six groups (n = 10) using different dentifrice treatments: placebo; nano-HA without F ; 1,100 µg F /g; 1,100 µg F /g + nano-HA; 5,000 µg F /g; and 5,000 µg F /g + nano-HA. A pH cycling model was performed for 10 days, in which treatments were performed twice a day. After that period, the longitudinal hardness was evaluated and the area of demineralization (ΔS) was calculated. The formulated dentifrices were evaluated for primary stability, cytotoxicity, and other technical parameters. Two-way ANOVA and Tukey's test with p set at 5% were used for data analysis. The experimental dentifrices were stable and had no cytotoxicity. Regarding dentin demineralization, the placebo group significantly increased ΔS compared to all other treatment groups (p<0.001). The dentifrices containing 5,000 µg F /g, regardless of the presence of nano-HA, led to a smaller lesion area in relation to the other treatments (p<0.001). The findings of this study suggest that nano-HA reduced dentin demineralization, and dentifrice with 5,000 µg F /g dentifrices, regardless of the presence of nano-HA, showed a greater reduction in root dentin demineralization.

The cariogenicity of starch alone or in combination with sucrose is controversial and the effect on dentine demineralization and on the dental biofilm formed has not been explored under controlled conditions. A crossover, single-blind study was conducted in four steps of 14 days each, during which 11 volunteers wore palatal appliance containing 10 slabs of root dentine to which the following treatments were applied extraorally: 2% starch gel-like solution (starch group); 10% sucrose solution (sucrose group); a solution containing 2% starch and 10% sucrose (starch + sucrose group), or 2% starch solution followed by 10% sucrose solution (starch → sucrose group). On the 14th day of each phase the biofilms were collected for biochemical and microbiological analyses, and dentine demineralization was assessed by hardness. A higher demineralization was found in dentine exposed to sucrose and starch sucrose combinations than to starch alone (p < 0.01), but the sucrose-containing groups did not differ significantly from each other (p > 0.05). The concentrations of soluble and insoluble extracellular polysaccharides (EPS), and the proportion of insoluble EPS, were lower in the biofilm formed in presence of starch (p < 0.01) than in those formed in the presence of sucrose or sucrose/starch combinations; however, no significant difference was observed among the groups containing sucrose (p > 0.05). RNA was successfully isolated and purified from in situ biofilms and only biofilms formed in response to sucrose and starch/sucrose combinations showed detectable levels of gtfB and gtfC mRNA. The findings suggest that the combination of starch with sucrose may not be more cariogenic to dentine than sucrose alone.

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.

Fibroblast growth factor-2 (FGF-2) enhances the formation of new alveolar bone, cementum, and periodontal ligament (PDL) in periodontal defect models. However, the mechanism through which FGF-2 acts in periodontal regeneration in vivo has not been fully clarified yet. To reveal the action mechanism, the formation of regenerated tissue and gene expression at the early phase were analyzed in a beagle dog 3-wall periodontal defect model. FGF-2 (0.3%) or the vehicle (hydroxypropyl cellulose) only were topically applied to the defect in FGF-2 and control groups, respectively. Then, the amount of regenerated tissues and the number of proliferating cells at 3, 7, 14, and 28 days and the number of blood vessels at 7 days were quantitated histologically. Additionally, the expression of osteogenic genes in the regenerated tissue was evaluated by real-time PCR at 7 and 14 days. Compared with the control, cell proliferation around the existing bone and PDL, connective tissue formation on the root surface, and new bone formation in the defect at 7 days were significantly promoted by FGF-2. Additionally, the number of blood vessels at 7 days was increased by FGF-2 treatment. At 28 days, new cementum and PDL were extended by FGF-2. Moreover, FGF-2 increased the expression of bone morphogenetic protein 2 (BMP-2) and osteoblast differentiation markers (osterix, alkaline phosphatase, and osteocalcin) in the regenerated tissue. We revealed the facilitatory mechanisms of FGF-2 in periodontal regeneration in vivo. First, the proliferation of fibroblastic cells derived from bone marrow and PDL was accelerated and enhanced by FGF-2. Second, angiogenesis was enhanced by FGF-2 treatment. Finally, osteoblastic differentiation and bone formation, at least in part due to BMP-2 production, were rapidly induced by FGF-2. Therefore, these multifaceted effects of FGF-2 promote new tissue formation at the early regeneration phase, leading to enhanced formation of new bone, cementum, and PDL.

To investigate the effects of Epigallocatechin gallate (EGCG) and curcumin, as a final irrigant on the fracture resistance of irradiated root that obturated with an epoxy resin sealer. Eighty mandibular premolars were randomly divided into non-irradiated (NIR) and irradiated (IR) groups. The teeth were irradiated at 2 Gy per fraction, 5 times a week for a total dose of 60 Gy over 6 weeks. All specimens were decoronated, remaining 13±1 mm root length. Two groups were subdivided into four groups (n = 10): 1) non-instrumented; the intact root served as control. The other roots were instrumented with a pro-taper NiTi rotary system. The final irrigation used was 17% EDTA, followed by three irrigation solution groups; 2) 2.5% NaOCl, 3) 0.02% EGCG, and 4) 0.1% curcumin. Root canals were filled with gutta-percha and AH plus. All specimens were embedded in self-curing acrylic resin and loaded vertically at 1 mm/min until fracture occurred. Also, sealer penetration was assessed by confocal laser scanning microscopy (CLSM). The data were evaluated statistically using two-way ANOVA and Tukey test (α = 0.05). In irradiated roots, fracture resistance of EGCG and curcumin groups did not differ from non-instrumented roots, but they were higher than the NaOCl group (P = 0.006). However, NaOCl, EGCG, and curcumin in irradiated roots had comparable strength that was higher than in the non-instrumented group (p<0.001). Difference between irradiated and non-irradiated roots was observed only for NaOCl and non-instrumented groups (P≤0.004). In irradiated roots, a higher sealer penetration was observed in EGCG and curcumin groups compared to NaOCl. EGCG and curcumin could be promising final irrigants to reverse the adverse effect of radiotherapy on the strength of irradiated roots obturated with AH Plus sealer.

This crossover study aimed to investigate abrasion of previously eroded hard dental tissues by a whitening dentifrice compared to a regular dentifrice. After a 3-day lead-in period, 14 volunteers were randomly assigned to use one of the toothpastes while wearing a removable appliance, containing 3 enamel and 3 root dentine slabs on each side. On the first day salivary pellicle was allowed to form. Twice daily for the following 3 days, one side of each appliance was immersed in an acidic carbonated drink ex vivo while the other side remained unexposed. Specimens were then brushed with the allocated dentifrice. After a 3-day washout period, new sets of enamel and dentine slabs were mounted in the appliances and the participants commenced period 2 using the alternative toothpaste. Acid-treated specimens always showed more wear than untreated specimens. The whitening dentifrice did not significantly increase the wear of softened enamel compared with the regular dentifrice. Brushing with the whitening toothpaste led to significantly greater wear of sound enamel and of both eroded and sound dentine than the regular dentifrice. The results suggest that whitening dentifrices may not increase the wear of acid-softened enamel but may have a more deleterious effect on dentine than regular toothpastes.

Periodontal regeneration using EDTA or PDGF showed promising results, but the effect of combined application was still unclear. This study aimed to verify the effect of EDTA and/or PDGF application on root adhesion and proliferation of PDL fibroblast cells. Eighty specimens were prepared from forty periodontitis teeth and made five groups: (1) diseased (untreated), (2) SRP (scaling root planing), (3) EDTA (24%), (4) PDGF (25 ng/ml) and (5) Combined application of EDTA and PDGF. Periodontal ligament cells were cultured on the above conditioned dentin plate, and SEM examination was preformed and cells were counted within a representative standard area for both cell morphology and density. All groups including untreated showed significantly increase of adhered cells from baseline to 7 days. Among them, rate of increase was much higher in EDTA, PDGF, and combined groups. ANOVA test indicated that the number of cells in PDGF and combined groups was significantly higher than diseased group at 1 day. On day 7, PDGF and combined groups showed significantly higher number of adhesion cells than that found in the diseased, SRP and EDTA groups. Thus, root conditioning with EDTA enhanced cell adhesion more than SRP alone. There was no significant difference of cell number between PDGF and combined group. Combined application of EDTA and PDGF increased significantly PDL cell adhesion than EDTA alone. PDGF alone, however, also showed comparable effect to combined application at all periods. Thus, synergistic effect between PDGF and EDTA was not observed.

This study explores innovative passive ultrasonic irrigation (PUI) parameters by investigating a novel, shorter, and more repetitive agitation cycle to enhance the penetration depth of sodium hypochlorite (NaOCl) within dentinal tubules. Forty extracted human mandibular premolars were prepared (size 40, 0.06 taper) and stained with 0.5% crystal violet. Samples were divided into four irrigation groups: (I) Conventional Needle Irrigation (CNI), (II) PUI with 2 cycles of 30 s each, (III) PUI with 3 cycles of 20 s each, and (IV) PUI with 6 cycles of 10 s each. Coronal, middle, and apical sections were analyzed by light microscopy, and NaOCl penetration depth was measured at four sites (mesial, distal, buccal, lingual) using ImageJ. Data were analyzed using Kruskal-Wallis/Dunn’s test (α < 0.05). The CNI group demonstrated significantly lower NaOCl penetration compared to all PUI groups ( P  < 0.001). The 6 × 10 s protocol achieved significantly greater penetration in all regions ( P  < 0.001). While no significant differences were observed between the 2 × 30 s and 3 × 20 s protocols in the coronal and middle thirds, the 3 × 20 s group showed significantly greater penetration in the apical third ( P  < 0.001). The findings of this study suggest that optimizing PUI agitation parameters can lead to more effective, efficient, and predictable endodontic treatment, enhancing irrigation efficacy. Shorter and more frequent cycles (6 × 10 s) proved to be the most effective, providing valuable information for clinical practice.