中文摘要：

目的 探讨自体骨髓间充质干细胞（BMSCs）构建成的细胞膜片结合低温保存的自体骨瓣修复兔颅骨极限缺损的效果. 方法 采集兔新鲜骨髓进行BMSCs的分离培养,并应用维生素C培养液诱导其生长成细胞膜片,同时对颅骨缺损模型取下的自体骨瓣进行无菌-80℃低温保存.将12只新西兰大白兔按随机数字表法分为实验组和对照组（每组各6只）,其中实验组将细胞膜片结合冻存自体骨瓣回植于原缺损部位,对照组单纯植入冻存自体骨瓣.术后12周时对2组兔进行活体颅骨CT扫描,并颅骨取材HE染色观察组织形态学变化,以评价骨修复效果. 结果 实验组兔骨缝边缘愈合良好,形成致密骨质,质地较硬,回植骨瓣活动度极差,CT见颅骨全层高密度影;对照组兔骨缝愈合较差,可见明显软组织及未愈合缺口,质地较弱,回植骨瓣活动度较好,CT见骨缝处有明显低密度影.HE染色见实验组断端局部愈合,形成新的骨层结构,并可见其中分散的骨细胞及滋养血管;对照组未形成完整骨层. 结论 BMSCs膜片能更有效地促进冻存自体骨瓣与缺损边缘的愈合,修复颅骨缺损.

英文摘要：

Objective To explore a method by using frozen autologous cranial bone combined with bone mesenchymal stem cells （BMSCs）-sheet to repair critical-size calvarial defects in rabbits. Methods BMSCs were cultured in vitro and induced into BMSCs-shcets by cell culture with Vitamin C. Twelve New Zealand rabbits were randomly divided into experimental group and control group （n=6）; critical-size calvarial defect models in the two groups were established; the autogenous bone （diameter 15 mm） from the above critical-size calvarial defect rabbits was kept in -80 ℃ refrigerator. The rabbits in the experimental group were repaired by BMSCs-sheets and autologous cranial bone, while rabbits in control group were repaired only by autologous cranial bone. All rabbits were scaned by using competed tomography （CT） 12 weeks after transplantation and then sacrificed. Cranial defects were evaluated histologically and macroscopically. Results The rabbits in the experimental group showed a stronger potential in bone regeneration than those in the control group; the bone suture in the experimental group was better filled by dense bone, and had a harder texture as compared with those in the control group. CT scan showed high-density shadow in cranial defects of the experimental group, while low-density shadow was observed in CT scan of the control group. In experimental group, HE staining indicated partial healing in broken ends, formation of newly bone layer, dispersed bone cells, and nourishvessels; in contrast, the control group did not form a complete layer of bone. Conclusion Frozen autologous bone and cell-sheet combination is a promising strategy in improving the healing of large area bone defect.