成骨诱导后的脐血间充质干细胞与β-磷酸三钙复合植骨材料对兔颅骨缺损的修复作用  

Repairing of rabbit skull defect by combination of artificial bone graft formed by umbilical cord blood-derived mesenchymal stem cells induced by bony conditioned media with β-tricalcium phosphate

作　　者：郑德宇[1] 刘建生[1] 杨依勇[1] 秦书俭[1] 包翠芬[1] 姚素艳[2] 

机构地区：[1]辽宁医学院解剖学教研室,辽宁锦州121001 [2]辽宁医学院病理生理学教研室,辽宁锦州121001

出　　处：《西安交通大学学报（医学版）》2011年第4期441-444,476,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：辽宁省教育厅创新团队(No.2010T061);辽宁省教育厅青年基金(No.05L140)~~

摘　　要：目的探讨成骨诱导后的人脐血间充质干细胞(UCB-MSCs)与β-磷酸三钙(TCP)构成的人工植骨材料修复兔颅骨缺损的作用。方法在无菌条件下用密度梯度离心和贴壁培养法分离培养人UCB-MSCs;取传3代细胞应用条件培养基进行成骨诱导2周后,绘制细胞生长曲线并检测碱性磷酸酶(AKP)、骨钙素(OCN)和钙离子的含量;成骨诱导后UCB-MSCs与β-TCP复合培养后形成人工植骨材料,用扫描电镜观察成骨诱导后的UCB-MSCs在β-TCP表面的生长状况;按人工植骨材料的形状制作兔颅骨全层缺损模型,术后4周行颅骨X线摄片,分析人工材料对骨缺损的修复作用。结果采用Percoll(1.073 g/mL)密度梯度离心和贴壁培养得到的UCB-MSCs大小较为均匀、梭形或星形的成纤维细胞样细胞。成骨诱导前后细胞的生长曲线测定无明显变化;UCB-MSCs内AKP、培养基中的OCN含量和细胞内钙离子的含量与对照组相比均明显增高(P<0.01);成骨诱导的细胞在β-TCP表面生长良好,在细胞周围有白色的钙盐沉积。人工植骨材料植入骨缺损4周后,发现复合材料与颅骨缺损中间大部分被高密度影所充填。结论成骨诱导后的UCB-MSCs呈现成骨细胞特性,在β-TCP表面生长状态良好;与β-TCP构成人工植骨材料具有促进骨缺损修复的作用。Objective To explore the repairing function for rabbit skull defect of artificial bone graft formed by umbilical cord blood-derived mesenchymal stem cells（UCB-MSCs） induced by bony conditioned medium with β-tricalcium phosphate（TCP）.Methods The human UCB-MSCs were obtained by density gradient centrifugation method and adherent method in sterile condition.The UCB-MSCs of third-generation of propagation were induced by bony conditioned media.Two weeks later,the growth curve of induced human UCB-MSCs and the contents of AKP and Ca2＋ of the UCB-MSCs and OCN in the medium were detected.The biocompatibility of induced human UCB-MSCs with β-TCP was observed after the MSCs were transplanted onto the β-TCP.The artificial bone graft was obtained by this way.Then the bone graft was transplanted into the rabbit skull defect.Four weeks later,the repairing function of artificial bone graft was observed with the naked eyes and X-ray.Results The obtained MSCs were similar in size,and were spindle-shaped or star-shaped fibroblasts-liked cells.By the cell growth curve,there was no significant change before and after bone induction.The contents of AKP and Ca2＋ of the UCB-MSCs induced by bony-conditioned media and OCN in the medium were higher than those in the control cells（P〈0.01）.UCB-MSCs grew well on the surface of β-TCP,and white calcium particles could be observed around the inducedUCB-MSCs.The high-density images were fulfilled in the space between the artificial bone graft and the rabbit full-layer skull defect after the bone graft was transplanted into the skull defect after 4 weeks.Conclusion The characteristics of bone-induced UCB-MSCs are in accordance with those of osteoblasts.The biocompatibility of these cells is satisfactory with TCP in vitro.The artificial bone graft can accelerate bone repairing in vivo.

关 键 词：成骨诱导 脐血间充质干细胞 Β-磷酸三钙 人工植骨材料 骨缺损修复 

分 类 号：R392.12[医药卫生—免疫学]