Deng Yuan, Zhou Huifang, Gu Ping, Fan Xianqun
Department of Ophthalmology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Invest Ophthalmol Vis Sci. 2014 Aug 28;55(9):6016-23. doi: 10.1167/iovs.14-14977.
To investigate the role of miR-31 genetically modified bone marrow mesenchymal stem cells (BMSCs) composited with porous β-tricalcium phosphate (β-TCP) scaffolds in repairing canine medial orbital wall defects.
A circular bone defect (10 mm in diameter) was created on the canine medial orbital wall. After canine BMSCs were isolated and transfected with lentiviral vectors encoding miR-31, anti-miR-31 (anti-miR), and negative control (miR-Neg) in vitro, they were seeded onto porous β-TCP scaffolds and implanted to repair the orbital defects. Spiral computed tomography (CT) scans were conducted at 4 and 16 weeks after surgery. Micro-CT and histological analysis were performed at 16 weeks after surgery. The results were analyzed to evaluate the extent of bone repair.
Examination with CT revealed good recovery in the anti-miR group at 16 weeks after surgery. In addition, the micro-CT analysis showed that the bone mineral density and new bone volume increased in the anti-miR group and decreased in the miR-31 group compared with that in the miR-Neg group. Histologic analysis confirmed that the formation of new bone and extent of β-TCP degradation were enhanced in the anti-miR and attenuated in the miR-31 group. In situ hybridization and immunohistochemical analysis further confirmed the micro-CT findings.
The use of BMSCs with suppression of miR-31 expression combined with β-TCP scaffolds can efficiently repair medial orbital wall defects in dogs.
探讨基因修饰的微小RNA-31（miR-31）骨髓间充质干细胞（BMSCs）与多孔β-磷酸三钙（β-TCP）支架复合在修复犬眼眶内侧壁缺损中的作用。
在犬眼眶内侧壁制造一个直径10毫米的圆形骨缺损。犬BMSCs在体外分离后，分别用编码miR-31、抗miR-31（抗miR）和阴性对照（miR-Neg）的慢病毒载体转染，然后接种到多孔β-TCP支架上，植入以修复眼眶缺损。术后4周和16周进行螺旋计算机断层扫描（CT）。术后16周进行显微CT和组织学分析。分析结果以评估骨修复程度。
CT检查显示抗miR组在术后16周恢复良好。此外，显微CT分析表明，与miR-Neg组相比，抗miR组骨矿物质密度和新骨体积增加，miR-31组降低。组织学分析证实，抗miR组新骨形成和β-TCP降解程度增强，miR-31组减弱。原位杂交和免疫组化分析进一步证实了显微CT的结果。
使用抑制miR-31表达的BMSCs与β-TCP支架联合可有效修复犬眼眶内侧壁缺损。
