Cyclic Stretch Enhances Osteogenic Differentiation of Human Periodontal Ligament Cells via YAP Activation.

Periodontal remodeling and alveolar bone resorption and formation play essential roles during orthodontic tooth movement (OTM). In the process, human periodontal ligament cells (HPDLCs) sense and respond to orthodontic forces, contributing to the alveolar bone formation. However, the underlying mechanism in this process is not fully elucidated. In the present study, cyclic stress stimulus was applied on HPDLCs to mimic the orthodontic forces during OTM. Our results demonstrated that cyclic stretch promoted the osteogenic differentiation of HPDLCs. Moreover, our data suggested that yes-associated protein (YAP), the Hippo pathway effector, which also involved in mechanical signaling transduction, was activated as we found that the nuclear translocation of YAP was significantly increased in the cyclic stress treated HPDLCs. The mRNA expression of CTGF and CYR61, the target genes of YAP, was also remarkably increased. Furthermore, knockdown of YAP suppressed the cyclic stretch induced osteogenesis in HPDLCs, while overexpression of YAP in HPDLCs enhanced osteogenesis. We also noticed that YAP activities could be suppressed by the ROCK and nonmuscle myosin II inhibitors, Y-27632 and Blebbistatin. The inhibitors also significantly inhibited the cyclic stretch induced osteogenesis in HPDLCs. Finally, in the murine OTM model, our results revealed that YAP was upregulated and nuclearly translocated in the PDLCs at the tension side. In summary, our present study demonstrated that cytoskeleton remodeling induced activation of YAP signaling pathway was crucial for the cyclic stretch-induced osteogenesis of HPDLCs, which might play important roles during OTM.

Wnt5a mediated canonical Wnt signaling pathway activation in orthodontic tooth movement: possible role in the tension force-induced bone formation.

Orthodontic tooth movement (OTM) is associated with bone remodeling mediated by orthodontic mechanical loading. Increasing studies reported that Wnt signaling played crucial roles in mechanical stimuli induced bone remodeling. However, little is known about the involvement of Wnt signaling in orthodontic force-induced bone formation during OTM. In virtue of the OTM mice model as we previously reported, where new bone formation was determined by micro-CT and immunoreactivity of osteocalcin and osterix, we explored the activation of Wnt signaling pathway during OTM. Our results proved the nuclei translocation of ß-catenin, suggesting the activation of canonical Wnt signaling pathway in the periodontal ligament cells (PDLCs) near the alveolar bone at the tension site (TS). Moreover, the immunoreactivity of Wnt5a, but not Wnt3a in PDLCs indicated the activation of canonical Wnt pathway might be mediated by Wnt5a, but not Wnt3a as in most cases. The co-location of Wnt5a and ß-catenin that was evidenced by double labeling immunofluorescence staining further supported the hypothesis. In addition, the high expression of FZD4 and LRP5 in PDLCs at TS of periodontium suggested that the activation of Wnt signaling pathway was mediated by these receptors. The negligible expression of ROR2 also indicated that canonical but not non-canonical Wnt signaling pathway was activated by Wnt5a, since previous studies demonstrated that the activation of canonical/non-canonical Wnt signaling pathway was largely dependent on the receptors. In summary, we here reported that Wnt5a mediated activation of canonical Wnt signaling pathway might contribute to the orthodontic force induced bone remodeling.

Correlation Between Cephalometric Measures and End-of-Treatment Facial Attractiveness.

Sixty-nine experienced Chinese orthodontists evaluated 108 Chinese patients' facial attractiveness from set of photographs (frontal, lateral, and frontal smiling photos) taken at the end of orthodontic treatment. These 108 patients, which contained an equal number of patients with Class I, II, and III malocclusion, were randomly selected from 6 orthodontic treatment centers throughout China. Spearman rank-order correlation coefficients (rs) analyses were performed to examine agreement in ranking between all judge pairs. Pearson correlation and multivariate regression were performed to examine the correlation between cephalometric measures and end-of-treatment Photo Attractiveness Rank.96.68% judge pairs showed moderate correlated (+0.4 ≤ rs < +0.7) subjective rankings. Cephalometric measures significantly correlated with end-of-treatment Photo Attractiveness Rank included interincisal angle (r = 0.330, P < 0.05), L1/MP° (r = 0.386, P < 0.05), L1-NBmm (r = 0.451, P < 0.01), L1/NB° (r = 0.374, P < 0.05), and profile angle (r = 0.353, P < 0.05) in Class I patients with an explained variance of 32.8%, and ANB angle (r = 0.432, P < 0.01), angle of convexity (r = 0.448, P < 0.01), profile angle (r = 0.488, P < 0.01), Li to E-line (r = 0.374, P < 0.05), Li to B-line (r = 0.543, P < 0.01), and Z angle (r = 0.543, P < 0.01) in Class II patient with an explained variance of 43.3%.There was less association than expected between objective measurements on the lateral cephalograms and clinicians' rankings of facial attractiveness on clinical photography in Chinese patients. Straight-stand lower incisor was desired for facial attractiveness of Class I malocclusion; and sagittal relationship and lip prominence influence the esthetics of Class II malocclusion in Chinese population.

Mutation breeding of lycopene-producing strain Blakeslea trispora by a novel atmospheric and room temperature plasma (ARTP).

To improve the fermentation efficiency of lycopene, a plasma jet, driven by an active helium atom supplied with atmospheric and room temperature plasma (ARTP) biological breeding system, was used as a new method to generate mutations in Blakeslea trispora (-). After several rounds of screening, a mutant A5 with high concentration of lycopene and dry biomass was isolated, which showed a maximum lycopene concentration (26.4 ± 0.2 mg/g dry biomass) which was 55 % higher than the parent strain (16.9 ± 0.3 mg/g dry biomass) in the production of lycopene. Compared with parent strain, B. trispora A5 required less dissolved oxygen (10 % less than that of parent strain) to reach maximum concentration in a 5-L stirred tank reactor batch fermentation.

[Expression of cathepsin K and IL-6 mRNA in root-resorbing tissue during tooth movement in rats].

OBJECTIVE: To investigate the expression and the localization of Cathepsin K and IL-6 mRNA in root-resorbing tissue and to elucidate the molecular changes and mechanism of root resorption induced by tooth movement. METHODS: Rats were subject to experimental tooth movement to induce root resorption. In situ hybridization was performed to identify the cells in root-resorbing tissue that produced Cathepsin K or IL-6 the difference of CK mRNA or IL-6 mRNA expression between root resorption group and control group was calculated by t-test. RESULTS: Cathepsin K mRNA was highly and selectively expressed in multinuclear odontoclast and IL-6 mRNA expressed in fibroblast, osteoblast, osteocyte and cementoblast. The expression of Cathepsin K mRNA and IL-6 mRNA in root-resorbing tissue increased evidently compared with the normal periodontium. CONCLUSIONS: Odontoclast in the root-resorbing tissue expresses Cathepsin K mRNA that participates in proteolysis during root resorption. IL-6 plays a very important role in the root resorption as a multifunctional cytokine.

[In vitro cytotoxicity evaluation of comfort denture adhesive].

OBJECTIVE: The purpose of this study was to evaluate the in vitro cytotoxicity of novel Comfort denture adhesive (Comfort-DA), which was developed by the authors, to human oral fibroblasts (HOFs). METHODS: A sample of Comfort-DA was prepared and extracted in culturing medium to prepare the eluate. Then the eluate was diluted by culturing medium to 50% and 75% concentration for the assessment of cytotoxicity by tetrazolium bromide (MTT) colorimetric assay. Wells containing fresh medium alone were served as control. Cell viability was recorded by optical density after culturing in an atmosphere of 5% CO2 and 95% air at 37 degrees C for 2, 3 and 4 days, respectively. The viability of HOF cells was evaluated by MTT assay to investigate cell proliferation. Optical density (OD) was measured by a spectrophotometer at 490 nm. Then evaluating the cytotoxicity grade in test groups according to the means of cell proliferation. ANOVA was used to test the statistical significance. RESULTS: The statistical analysis of the results of MTT cytological assay indicated significant difference (P < 0.05) in OD (indicate cell viability) between all concentrations of Comfort-DA and the control at all incubation times. The results of cell proliferation percentage also showed that the cytotoxicity grade of tested material only displayed "0-2". CONCLUSION: The generally favorable in vitro cytotoxicity of the Comfort-DA formulations indicates that this product may be an efficacious denture adhesive.

Laboratory evaluation of a new denture adhesive.

OBJECTIVE: The ideal denture adhesive demonstrates a neutral or slightly basic pH, satisfactory bonding load, and minimal toxicity to the oral mucosa. The purpose of this study was to evaluate the properties of a new denture adhesive (Comfort-DA, developed by the authors). METHODS: Comfort-DA was evaluated in the laboratory to determine the clinical effect of pH, bonding strength, and cytotoxicity. Firstly, the material was diluted at 1.0, 2.0, 3.3, 5.0, and 10% concentrations. Then the pH values were measured at 1-, 2-, 3-, 4-, 5-, 6-, and 8-hour intervals. Secondly, bonding load test was performed between acrylic resin samples on a universal testing machine. An existing product, Fittydent, was used as a control. Finally, the cytotoxicity of several dilutions of Comfort-DA was recorded with a MTT assay on human oral fibroblast cells. Cell viability was recorded by optical density after culturing in an atmosphere of 5% CO2 and 95% air at 37 degrees C for 2, 3 and 4 days, respectively. Wells containing fresh medium alone served as controls. All the results were compared with ANOVA or Tukey intervals (alpha = 0.05). RESULTS: Comfort-DA remained above the critical pH for hydroxyapatite (pH 6.5) in all dilutions throughout the study. Results of bonding strength test indicated that Comfort-DA was significantly stronger than Fittydent (p < 0.05). The statistical analysis of results for cytotoxicity showed that Comfort-DA only displayed slight cytotoxicity. SIGNIFICANCE: The generally favorable laboratory performance of Comfort-DA formulations indicates that this product may be an efficacious denture adhesive.