Short-term evaluation of photobiomodulation therapy on the proliferation and undifferentiated status of dental pulp stem cells.

The aim of this in vitro study was to analyze the effect of photobiomodulation therapy (PBMT) on the proliferation and undifferentiating status of stem cell from human exfoliated deciduous teeth (SHEDs). PBMT was carried out with an aluminum gallium indium phosphide (InGaAlP) diode laser in contact and punctual mode (continuous wave, 660 nm, 20 mW, 0.028 cm2, and average energy densities of 1 (1 s), 3 (4 s), 5 (7 s), 10 (14 s), 15 (21 s), or 20 (28 s) J/cm2 per point). The immunoprofile of the SHEDs was analyzed using flow cytometry. Cell proliferation was assessed by the MTT reduction assay. Gene expressions of mesenchymal stem cell markers (OCT4, Nestin, CD90, and CD105) were assessed by RT-qPCR 48 h after PBMT. Data were compared by analysis of variance (ANOVA) and Tukey's test (p ≤ 0.05). Cells cultured under nutritional deficit and treated with PBMT at 5 J/cm2 presented similar cell growth than those of positive control group. Cell growth was significantly higher than those of other groups. Mesenchymal stem cell gene markers were still expressed after PBMT at 5 J/cm2. In a short-term analysis, PBMT increases the number of stem cells with no interference in the undifferentiated state of the irradiated cells, which opens wide possibilities for application in tissue regeneration.

Cytotoxicity, genotoxicity and antibacterial effectiveness of a bioceramic endodontic sealer.

AIM: To compare the characteristics of bioceramic endodontic sealer Endosequence BC sealer with those of AH Plus sealer. METHODOLOGY: Cytotoxicity and genotoxicity were analysed on human gingival fibroblasts submitted to cell culture medium conditioned by sealers using the MTT reduction assay and micronucleus formation test (MNT), respectively. Cells grown on fresh medium served as controls. Cell viabilities were measured at 1, 3, 5 and 7 days. The antibacterial activity was analysed on an Enterococcus faecalis strain (ATCC 29212) using both on agar diffusion test (ADT) and a direct contact test (DCT). The inhibition zones in ADT were measured after 48 h and the colony-forming units counting in the DCT after 1, 24, 72 and 168 h. Data were compared by anova and Tukey's test and MNT by Fisher's exact test (P < 0.05). RESULTS: Cultures submitted to Endosequence BC sealer had a significantly higher number of viable cells (P < 0.01) and less micronucleus formation (P < 0.05) than AH Plus sealer. Endosequence BC sealer exhibited significantly smaller inhibition zones (6.00 ± 0.03 mm) than AH Plus sealer (10.31 ± 0.21 mm) (P < 0.05). Moreover, Endosequence BC sealer had significantly smaller antibacterial activity than AH Plus sealer up to 1 h of direct contact (P < 0.05). On other exposure times, both materials had similar antibacterial effectiveness (P > 0.05). CONCLUSIONS: Bioceramic-based sealer had less cytotoxicity and genotoxicity and similar antibacterial effect against E. faecalis in comparison with AH Plus sealer.

Quantification of peroxide ion passage in dentin, enamel, and cementum after internal bleaching with hydrogen peroxide.

The aim of this study was to evaluate the amount of peroxide passage from the pulp chamber to the external enamel surface during the internal bleaching technique. Fifty bovine teeth were sectioned transversally 5 mm below the cemento-enamel junction (CEJ), and the remaining part of the root was sealed with a 2-mm layer of glass ionomer cement. The external surface of the samples was coated with nail varnish, with the exception of standardized circular areas (6-mm diameter) located on the enamel, exposed dentin, or cementum surface of the tooth. The teeth were divided into three experimental groups according to exposed areas close to the CEJ and into two control groups (n=10/group), as follows: GE, enamel exposure area; GC, cementum exposed area; GD, dentin exposed area; Negative control, no presence of internal bleaching agent and uncoated surface; and Positive control, pulp chamber filled with bleaching agent and external surface totally coated with nail varnish. The pulp chamber was filled with 35% hydrogen peroxide (Opalescence Endo, Ultradent). Each sample was placed inside of individual flasks with 1000 µL of acetate buffer solution, 2 M (pH 4.5). After seven days, the buffer solution was transferred to a glass tube, in which 100 µL of leuco-crystal violet and 50 µL of horseradish peroxidase were added, producing a blue solution. The optical density of the blue solution was determined by spectrophotometer and converted into microgram equivalents of hydrogen peroxide. Data were submitted to Kruskal-Wallis and Dunn-Bonferroni tests (α=0.05). All experimental groups presented passage of peroxide to the external surface that was statistically different from that observed in the control groups. It was verified that the passage of peroxide was higher in GD than in GE (p<0.01). The GC group presented a significantly lower peroxide passage than did GD and GE (p<0.01). It can be concluded that the hydrogen peroxide placed into the pulp chamber passed through the dental hard tissues, reaching the external surface and the periodontal tissue. The cementum surface was less permeable than were the dentin and enamel surfaces.

Ex vivo performance of five methods for root canal length determination in primary anterior teeth.

AIM: To evaluate in a laboratory setting the performance of five methods for the determination of root canal length in primary anterior teeth. METHODOLOGY: Twenty extracted primary incisors, with at least two-thirds of the root, were used. After access cavity preparation, the teeth were embedded in alginate mixed with 0.9% sodium chloride solution. One operator determined root canal length using tactile sense (T), conventional radiography (RAD), tactile sense and conventional radiography (T + RAD), digital radiography (RDIG) and Root ZX electronic apex locator (EAL) methods. Next, the actual length (AL) was visually determined using a K-file from the coronal reference to the apical foramen or apical resorption level. The measurements obtained through each method were compared to the AL using the intraclass correlation coefficient (ICC) with the limits of agreement calculated with Bland and Altman analysis. The measurements were classified as acceptable (+/-1 mm from the AL) or not (>1 mm shorter or longer), and the McNemar test was employed for method comparison. RESULTS: Differences, limits of agreement and ICCs for each method were respectively EAL = -0.29; -1.02 to 0.44; 0.990; T + RAD = 0.17; -2.18 to 2.51; 0.929; RAD = 0.50; -3.41 to 4.41; 0.818; RDIG = 0.95; -3.76 to 5.65; 0.700; and T = -0.48; -5.59 to 4.64; 0.499. The most accurate and acceptable method was the EAL, followed by the T + RAD. : The EAL method performed best for root canal length determination in primary teeth.