以β-磷酸三钙为支架组织工程骨在兔脊柱后外侧融合中的作用研究

Effects of tissue engineering bone constructed by β-tricalcium phosphate scaffold on rabbits posterolateral spinal fusion

目的探索兔骨髓基质干细胞﹙Bone marrow stromal cells,BMSCs﹚与-磷酸三钙﹙β-tricalcium phosphate,β-TCP﹚复合的生物相容性以及该组织工程骨的成骨效果。方法新西兰大白兔12只,雌雄不限,体重3±0.25kg。于L4∕5节段棘突两侧行脊柱融合手术,随机将2种材料﹙①单纯材料组:-TCP颗粒;②组织工程骨组:将BMSCs接种到-TCP材料上静态培养24小时﹚均匀植入打磨好的骨槽内。术后12周处死。利用MTT检测、扫描电镜、荧光染色等方式观察细胞的粘附及伸展情况,利用X线、显微-CT、组织学观察材料降解及骨长入情况。结果通过MTT、扫描电镜等方法观察发现BMSCs与-TCP复合24小时后,多孔的-TCP表面和内部均可见大量细胞粘附,细胞伸展和生长良好;术后12周影像学和组织学研究发现,组织工程骨组有明显的新生骨生成,而单纯材料组没有,2组材料均无明显降解。结论该组织工程骨无论在体内外均具有良好的生物相容性,与单纯-TCP相比能够更好、更快的成骨,具有更广阔的应用前景。

Objective To investigate the biocompatibility of bone marrow stem cells combined with-tricalcium phosphate and the osteogenesis of this tissue engineering bone in the early stage of recovery.Method Posterolateral spinal fusion was carried out in 12 White New Zealand rabbits using one of the following graft materials in the bone solt ： ①β-tricalcium phosphate alone;②β-tricalcium phosphate combined with bone marrow stem cells after static culture for 24 hours.Activities of BMSCs were evaluated by MTT,Scanning electron microscopy and fluorescence staining.All the rabbits were killed 12 weeks after surgery to evaluate with radiography,Micro-CT,histologic analysis.Result 24h after inoculation,there were large number of BMSCs on the surface and in the pore of the scaffold material by Scanning electron microscopy and fluorescence staining.Cells which adhered to the surface and the pole,growed and expanded very well.New bone was observed in the tissue engineering bone group obviously by radiography,Micro-CT,histologic analysis.However,almost no new bone can be find in theβ-tricalcium phosphate alone group.Conclusion The tissue engineering bone not only has a good biocompatibility,but also has the capacity to form new bone earlier and better compare withβ-tricalcium phosphate alone.Therefore,this tissue engineering bone has a better application prospect.