[Effects of let-7a on proliferation, osteogenic differentiation and apoptosis of human dental pulp stem cells].

PURPOSE: To study the effect and possible mechanism of let-7a on proliferation, differentiation and apoptosis of human dental pulp stem cell (hDPSCs). METHODS: The cells were divided into four groups: overexpression control (let-7a control/let-7a agomir control), overexpression let-7a (let-7a mimics/let-7a agomir), knockdown let-7a control (let-7a inhibitor control) and knockdown let-7a (let-7a inhibitor). Cell counting kit-8 assay(CCK-8) was used to detect the proliferation of cells at 24 hours, 48 hours and 72 hours after transfection. Calcified nodules were detected by Alizarin red staining. The protein expression of alkaline phosphatase (ALP), osteopontin (OPN), 4E-binding protein 1 (4EBP1), p-4EBP1, mammalian target of rapamycin (mTOR) and p-mTOR were detected by Western blot. Annexin V-APC/7-AAD cell apoptosis detection kit was used to detect the level of apoptosis after transfection. Statistical analysis was performed using GraphPad Prism 5.0 software. RESULTS: Let-7a inhibited proliferation of hDPSCs and promoted odontoblast differentiation and apoptosis. Let-7a down-regulated the expression of 4EBP1, p-4EBP1, mTOR and p-mTOR. CONCLUSIONS: Let-7a may inhibit proliferation of hDPSCs and promote their differentiation and apoptosis by inhibiting mTOR-4EBP1 molecular pathway.

Effectiveness of the attachment position in molar intrusion with clear aligners: a finite element study.

OBJECTIVE: To evaluate the biomechanical effects of different attachments' position for maxillary molar intrusion with clear aligner treatment by finite element analysis. METHODS: Cone-beam computed tomography images of a patient with supra-eruption of the maxillary second molars were selected to construct three-dimensional models of the maxilla, periodontal ligaments, dentition, and clear aligner. The models were divided into four groups depending on the attachment location on the first molar: (1) no attachment (NA), (2) buccal attachment (BA), (3) palatal attachment (PA), and (4) bucco-palatal attachment (BPA). After applying an intrusion of 0.2 mm on the second molar, displacements and stress distributions of the teeth, aligner, and periodontal ligament were analyzed with the finite element software. RESULTS: All groups displayed equivalent movement patterns of aligners. The NA and BA groups showed buccal tipping of the second molar, while the PA group showed palatal tipping. The BPA group had the highest intruding value and the lowest buccal/palatal tipping value. All groups showed mesial tipping of the second molar. Stress distribution in the periodontal ligament strongly correlated with the attachment position. The BPA group showed the best stress distribution. CONCLUSION: Combined BA and PA could effectively prevent buccal and palatal tipping and showed the best efficiency in intruding the second molar. The second molar showed an unavoidable tendency to tip mesially, regardless of the attachment position.

Stem Cells from Human Exfoliated Deciduous Teeth (SHEDs) and Dental Pulp Stem Cells (DPSCs) Display a Similar Profile with Pericytes.

BACKGROUND: Pericytes play an important role in forming functional blood vessels and establishing stable and effective microcirculation, which is crucial for vascular tissue engineering. The slow ex vivo expansion rate, limited proliferative capacity, and variability of tissue-specific phenotypes would hinder experimental studies and clinical translation of primary pericytes. In this study, the angiogenic and pericyte functions of stem cells from human exfoliated deciduous teeth (SHEDs) and postnatal human dental pulp stem cells (DPSCs) were investigated. METHODS: Osteogenic and adipogenic induction assays were performed to evaluate the mesenchymal potential of SHEDs, DPSCs, and pericytes. An in vitro Matrigel angiogenesis assay was conducted to reveal the ability of SHEDs, DPSCs, and pericytes to stabilize vascular-like structures. Quantitative real-time polymerase chain reaction (RT-qPCR) was performed to evaluate mRNA expression. Flow cytometry, western blotting, and immunostaining were used to assess the protein expression. Wound healing and transwell assays were performed to evaluate the migration ability of SHEDs, DPSCs, and pericytes. RESULTS: The osteogenic and adipogenic induction assays showed that SHEDs, DPSCs, and pericytes exhibited similar stem cell characteristics. The mRNA expression levels of PDGFR-ß, α-SMA, NG2, and DEMSIN in SHEDs and DPSCs cultured in EC medium were significantly higher than those in the control groups on day 7 (P < 0.05), but significantly higher than those in the pericytes group on day 14 (P < 0.05). Flow cytometry showed that high proportions of SHEDs and DPSCs were positive for various pericyte markers on day 7. The DPSCs, SHEDs, and pericytes displayed strong migration ability; however, there was no significant difference among the groups (P > 0.05). CONCLUSION: The SHEDs and DPSCs display a profile similar to that of pericytes. Our study lays a solid theoretical foundation for the clinical use of dental pulp stem cells as a potential candidate to replace pericytes.

[Effect of transfected EphB4 on osteogenic differentiation in stem cells from human exfoliated deciduous teeth].

PURPOSE: To evaluate the effect of stem cells from human exfoliated deciduous teeth (SHED) transfected with EphB4 gene in regulating osteogenic differentiation. METHODS: Human dental pulp tissue were harvested from extracted deciduous teeth and digested by collagenase and dispase. The SHEDs were transfected with transgenic (hEphB4-GFP) vector or empty vector (GFP-vector). Real time-polymerase chain reaction（real time-PCR） analysis and Western blot were used to detect the expression of EphB4 in SHEDs after transfection. EphB4-SHEDs and GFP-SHEDs were subjected to osteogenic induction and assessed by alkaline phosphatase(ALP) assay and Alizarin-red S staining. SPSS 16.0 software package was used for statistical analysis. RESULTS: Real time-PCR revealed that the expression of EphB4 was significantly enhanced in EphB4-SHEDs compared to GFP-SHEDs (P<0.05). The expression of EphB4 protein was significantly higher (P<0.05) in EphB4-SHEDs compared to GFP-SHEDs. ALP assay and Alizarin-red S staining demonstrated higher ALP activity and increased mineralization with EphB4-SHEDs. CONCLUSIONS: The results indicate that transgenic expression of EphB4 in SHEDs could promote osteogenic differentiation.

[The mechanism of corticotomy accelerating orthodontic tooth movement in SD rats].

PURPOSE: To study the influences of corticotomy on orthodontic tooth movement (OTM) and the underlying mechanism in rats. METHODS: Forty-eight female Sprague-Dawley rats were randomly assigned to corticotomy group (Cort, 24 rats) or sham-corticotomy group (Sham, 24 rats). All rats were subjected OTM after corticotomy or sham surgery. 6 rats of each group were sacrificed at 0, 1, 3 and 7 day of OTM. OTM were measured with an electronic digital caliper. Osteoclasts were counted in pressure side with TRAP. RANKL were measure by IHC at pressure side. The data were analyzed with SPSS 16.0 software package. RESULTS: OTM at the 1 and 7 day in Cort group increased compared with sham group. Number of TRAP positive osteoclasts in pressure side increased in Cort group at the 3 and 7 day. Expression of RANKL in pressure side also increased in Cort group at the 3 and 7 day. CONCLUSIONS: Corticotomy accelerates OTM in rats and it may result from promoted bone resorption via increased RANKL expression in periodontal tissue.

Morphological analysis of apical foramen over-instrumented by three rotary NiTi systems.

OBJECTIVE: To evaluate morphological changes of the apical surface after root canal preparation with 1 mm beyond the apical foramen using ProTaper Universal (PTU) files, K3 files and Twisted files (TF), respectively. METHODS: Seventy teeth with a centered apical foramen and 70 teeth with a deviated apical foramen were included as group A and group B respectively. In each group, 20 teeth were randomly assigned for root canal preparation with PTU, K3 and TF files, respectively; the remaining 10 teeth were used as the control group without any preparation. The apical foramens were examined with scanning electronic microscopy. The foramen integrity damage (FID) and dentin defects (DDs) were noted and compared between different groups. RESULTS: FID and DD were significantly less in Group A. DDs was not found in the control group. Preparation with PTU, K3, and TF files caused FID in 6.67%, 10%, and 3.33% of teeth in the group A, and in 20%, 26.67%, and 10% in Group B, respectively. Preparation with PTU, K3, and TF files caused DD in 6.67%, 6.67%, and 3.33% of teeth in Group A, and in 23.33%, 26.67%, and 6.67% in Group B, respectively. PTU and K3 files produced more DDS than TF files. However, no significant difference was found between groups using PTU and K3 files. CONCLUSION: Rotary instrumentation caused less damage on the apical surface in foramencentered root canals than foramen-deviated root canals when working beyond the canal length. TF files had a tendency to produce less DDS compared with PTU or K3 files during over-instrumented root canals.

[Coculture of stem cells from the apical papilla and human umbilical vein endothelial cells enhances the angiogenic potential of dental pulp tissues].

PURPOSE: To observe the effect of coculture of stem cells from the apical papilla (SCAPs) and human umbilical vein endothelial cells (HUVECs) on the angiogenic potential of dental pulp tissues. METHODS: SCAPs were incubated in osteo/odontogenic, adipogenic, neurogenic induction medium and α-MEM medium, whose multilineage differentiation capacities were confirmed using alizarin red staining, oil red O staining and ßIII-tubulin immunofluorescent staining. The tubular length, branching points number and junctional areas were detected after 3, 6, 9 h since cells were seeded onto matrigel, and the data was analyzed using SPSS 16.0 software package. RESULTS: SCAPs in the experimental groups were detected having more lipid droplets, mineralization nodules and neuron-like cells. Coculture of SCAPs and HUVECs formed more vessel-like structures in tubular formation assay. CONCLUSIONS: SCAPs are capable of differentiating into fat, bone, and nerve-like cells in vitro. Coculture of SCAPs and HUVECs can enhance the angiogenic potential of dental pulp tissues.

[Treatment of primary trigeminal neuralgia with radiofrequency thermocoagulation: report of 648 consecutive cases].

PURPOSE: To analyze the clinical efficacy of radiofrequency thermocoagulation in the treatment of idiopathic trigeminal neuralgia, and discuss the method, skill of radiofrequency thermocoagulation and complications. METHODS: 648 patients with idiopathic trigeminal neuralgia , who were treated by radiofrequency thermocoagulation via foramen infraorbitale approach, lateral approach, anterior approach and other approach from July 2001 to March 2011 in our hospital, were observed and the clinical efficacy was evaluated. RESULTS: After the first treatment of the 648 patients, the rate of pain control was 98.3% via foramen infraorbitale approach, 91.0% via lateral approach and 95.5% via former approach. The overall response rate was 96.0%. 395 patients were followed up from 6 months to 2 years. The recurrent rate within one year was 9.6%, 20.5% within two years. Good response was achieved after re-treatment with radiofrequency thermocoagulation in recurrent patients. CONCLUSIONS: The clinical efficacy of radiofrequency thermocoagulation in the treatment of idiopathic trigeminal neuralgia is good and reliable. The operation is simple, the indication is wide, and the complication is fewer. CT location can improve the accuracy of puncture, and reduce complications.