[Osteogenic Capacity and Mettl14 and Notch1 Expression of Adipose-Derived Stem Cells from Osteoporotic Rats].

OBJECTIVE: To investigate the differences in the osteogenic capacity of osteoporotic adipose-derived stem cells (OP-ASCs) and normal control adipose-derived stem cells (Ctrl-ASCs), and to examine the expression levels of RNA methyltransferase like 14 (Mettl14) and the Notch signaling molecule 1 (Notch1). METHODS: The osteoporosis (OP) model of SD rats was established with ovariectomy (OVX). Micro-CT, HE staining and Masson staining were performed to identify the successful establishment of the OP model, OP-ASCs and Ctrl-ASCs were isolated and cultured adherently. Then, the three-way differentiation capacity of the adipose-derived stem cells (ASCs) was determined through alizarin red staining, alcian blue staining and oil red O staining and flow cytometry was conducted to examine the surface antigens CD29, CD44, CD90, CD31, CD34, and CD45. Alizarin red staining and comparison of the mRNA and protein expression of Run-related transcription factor 2 (Runx2) were done to explore the differences in osteogenic potential of OP-ASCs and Ctrl-ASCs. Real-time PCR and Western blot were performed to explore the expression differences of Mettl14 and Notch1 at mRNA and protein levels between OP-ASCs and Ctrl-ASCs. RESULTS: Micro-CT, HE and Masson staining results showed that the number of trabecular bone decreased and the spacing increased in the tibias of the osteoporosis group (OP group) compared with those of the control group (Ctrl group), indicating that the OP model was established successfully. Three-way differentiation and flow cytometry results confirmed the successful isolation and culture of ASCs. After osteogenic induction, alizarin red staining showed that OP-ASCs had fewer number and more scattered distribution of mineralized nodules than Ctrl-ASCs did. The expression of Runx2 in OP-ASCs was lower than that in Ctrl-ASCs ( P<0.05). Mettl14 as well as Notch1 showed lower expression in OP-ASCs than they did in Ctrl-ASCs ( P<0.05). CONCLUSION: The osteogenic capacity of OP-ASCs was lower compared with that of Ctrl-ASCs, Mettl14 expression of OP-ASCs was decreased compared with that of Ctrl-ASCs, and the Notch signaling pathway was inhibited in OP-ASCs. The study helps build the foundation for further investigation in the specific mechanisms of Mettl14 and Notch1 during osteogenic differentiation of OP-ASCs.

[Expression profiling of peroxisome proliferation activate receptor gamma2 and phosphorylation during adipogenic differentiation of adipose-derived stem cells].

OBJECTIVE: To investigate PPARgamma2 expression and phosphorylation involved in adipogenic differentiations of Adipose-derived Stem Cells (ASCs) with an aim to reveal the possible mechanisms of adipose development. METHODS: ASCs were obtained from the inguinal fat pads of GFP (green fluorescent protein) mice. The primary cells were cultured in histiocytic attachment cultivation. Adipogenic differentiation of the third generation of ASCs was induced with the conditional medium (alpha-MEM medium supplemented with 100 mL/L fetal bovine serum, 10(-6) mol/L dexamethasone, 10(-5) mol/L insulin, 0. 2 mmol/L indomethacin, and 0.5 mmol/L 3-isobutyl-1-methylxanthine). The levels of PPARgamma2 mRNA and the PPARgamma2 expression and phosphorylation at day 0, 1, 3, 5, 7, and 9 of induction were detected using RT-PCR, fluorescence-immunocytochemistry and western blot, respectively. RESULTS: The expression of PPARgamma2 and phosphorylation were significantly up-regulated during adipogenic differentiation of ASCs. CONCLUSION: PPARgamma2 is the key regulator of adipose development. The regulation of PPARgamma2 phosphorylation may promote adipogenic differentiation of ASCs.

[Application of a modified paramedian lower lip-submandibular approach in maxillary (subtotal) total resection].

OBJECTIVE: To investigate the clinical efficacy of a modified paramedian lower lip-submandibular approach for maxillary (subtotal) total resection. METHODS: Eleven patients of maxillary tumors underwent maxillary (subtotal) total resection through the modified paramedian lower lip-submandibular approach. Clinical follow-up visits were conducted to evaluate appearance restoration, facial nerve functional status, parotid gland functional status, and orbital region complication. RESULTS: During the follow-up period of 6-36 months, the appearance of all 11 patients recovered well. All cases presented hidden scars. No facial nerve and parotid duct injury, lower eyelid edema, lower eyelid ectropion, or epiphora in all cases was observed. CONCLUSIONS: Applying modified paramedian lower lip-submandibular approach to maxillary (subtotal) total resection effectively reduces incidence of orbital region complications including lower eyelid edema, lower eyelid ectropion, and epiphora, which often occur to traditional approach. The modified approach produces more subtle scars than other methods and should be applied to treatment of maxillary (subtotal) total resection.

[Precise investigation of digital guide plates applied to implant surgery of anterior teeth].

OBJECTIVE: To study the precision of digital guide plates applied to the implant surgery of anterior teeth. METHODS: Fifty patients scheduled to receive implant restoration treatment in anterior teeth were enrolled in this study and divided into two groups (n=25, each group): those who were given routine implant restoration treatment (control group, 45 implants) and those who received implant restoration treatment using a digital guide plate (test group, 51 implants). After implantation, planned and placed implants were superimposed using digital software, and deviations (corona, apex, depth, degree) were analyzed. Esthetic parameters were assessed at 1 week (baseline), 6 month, and 1 year post final restoration. Pink esthetic (PES) and white esthetic (WES) scores were respectively used to evaluate the soft tissue and restoration esthetic outcome. RESULTS: The deviation parameters in the test group were significantly lower than those in the control group (P<0.05). PES and WES values recorded for the control group at 1 week, 6 month, and 1 year post final restoration were significantly lower than those in the test group (P<0.05). CONCLUSIONS: The digital guide plate can improve the accuracy of the three-dimensional position of implants in the maxillary esthetic zone. As such, this device may play an important role in obtaining the ideal aesthetic effects of maxillary anterior teeth.

[Construction of recombinant gene adenovirus encoding enhanced green fluorecence protein-peroxisome proliferator-activated receptor gamma2 fusion protein and its expression in bone marrow mesenchymal stem cells].

OBJECTIVE: To construct mouse enhanced green fluorecence protein (EGFP) -peroxisome proliferator-activated receptor (PPAR)gamma2, and to detect EGFP-PPARgamma2 expression in infected mouse bone marrow mesenchymal stem cells (BMSC). METHODS: Cut the fragment of PPARgamma2 from the expression plasmid pcDNA flag PPARgamma2, then cloned the gene fragment into pEGFP-C1 and pEGFP-N1 vector. Subsequently, subclone the fragment EGFP-PPARgamma2 from pEGFP-C1-PPARgamma2 into the shuttle plasmid DC315. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid DC315-EGFP-PPARgamma2 and large adenovirus helper plasmid pBHGlox deltaE1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus Ad-EGFP-PPARgamma2 was confirmed. Then it was propagated in HEK293 cells. After the BMSC were transfected for 72 h, adipogenic differentiation was demonstrated. RESULTS: HEK293 cells were transfected with the pEGFP-C1-PPARgamma2 or pEGFP-N1-PPARgamma2 in mediation of liposome. The former green fluorescence protein was better than the latter by fluorescence microscope. The recombinant plasmids were digested and identified. Western blot analysis showed the expression of EGFP-PPARgamma2 in vitro. EGFP-PPARgamma2 protein was detectable in the nucleus of BMSC. CONCLUSION: The recombinant adenovirus encoding EGFP-PPARgamma2 fusion protein was successfully constructed, which provided a basis for application of EGFP-PPARgamma2 gene to adenovirus-mediated gene therapy.

[Development and application of a three-dimensional craniomaxillofacial measurement instrument].

OBJECTIVE: To evaluate the clinical application effect of a three-dimensional craniomaxillofacial measurement instrument. METHODS: A three-dimensional craniomaxillofacial measurement instrument was developed. Twelve patients of unilateral fracture of zygoma complex were treated with the help of the three-dimensional craniomaxillofacial measurement instrument. RESULTS: The therapeutic effects of twelve patients were satisfactory with the three-dimensional craniomaxillofacial measurement instrument. No complication occurred, such as infection, injury of nerves and veins. CONCLUSION: Three-dimensional craniomaxillofacial measurement instrument can exactly measure craniomaxillofacial hard tissue, and has adjuvant effect to restore and fix the fracture of unilateral zygoma complex.

[Clinical analysis of midfacial fractures with orbital floor fractures].

OBJECTIVE: To study the characters of the diagnosis and treatment for midfacial fractures with orbital floor fractures. METHODS: 136 cases of midfacial fractures with orbital floor fractures were diagnosed and treated. All patients underwent CT scans and accepted surgical intervention. Orbital floor fracture was restituted in 49 cases. Orbital floor defects were reconstructed with autogenous bone, titanium mesh or porous polyethylene implant (Medpor) in 21 cases. RESULTS: 136 cases recovered well, their facial appearance and function were improved significantly. There were no severe complications happened postoperatively in all cases. 2 cases had postoperative wounds infection and 1 case had temporary ablepsia, but healed completely. CONCLUSION: CT is the best method for diagnosing orbital floor fractures. The objective of treatment is restoration of normal orbital floor and orbital capacity, the reduction of orbital contents prolapsed and reconstruction of orbital floor fractures with repair materials.