摘要
[目的]探讨体外不同程度诱导人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,h UC-MSCs)成骨分化对大鼠股骨骨折局部骨形态发生蛋白2(bone morphogenetic protein 2,BMP-2)表达的影响。[方法](1)选用120只成年Wistar大鼠制作股骨不全骨折模型,随机分为A、B、C、D、E、F六组;(2)利用组织块贴壁法分离培养h UC-MSCs,流式细胞仪鉴定细胞表面抗原,给予体外成骨分化诱导,采用茜素红S染色鉴定,将未诱导和诱导3、7、11、15 d的h UC-MSCs分别注入A、B、C、D、E组实验动物骨折处,F组注射生理盐水作为空白对照组,于移植后1、2、3、4周分别切取标本,采用免疫组化显示BMP-2并行统计学分析。同时检测实验大鼠血常规。[结果]h UC-MSCs成骨诱导3周时细胞内出现大量钙结节。细胞移植各组BMP-2表达在第1、2、3、4周时均明显强于空白对照组。A、B、C、D、E各组BMP-2表达在第1周和第4周时没有显著性差异,在第2、3周时B、C两组显著强于A、D、E三组。BMP-2表达在第2周时出现最高峰,之后下降,在第4周时仅少量表达。六组之间血常规差异无统计学意义。[结论]h UC-MSCs具有较好的免疫相容性,移植前体外诱导h UCMSCs成骨分化能提高骨折愈合早期BMP-2的表达,诱导3、7 d效果最佳。
[Objective]To investigate the effect of human umbilical cord mesenchymal stem cells( h UC-MSCs) in vitro osteogenesis differentiation on the expression of bone morphogenetic protein 2( BMP-2) in the rats' femur fracture location.[Method]( 1) The femoral insufficiency fracture models was made using 120 Wistar rats,and divided into six groups randomly:group A,B,C,D,E,F.(2) h UC-MSCs were cultured and identificated and h UC-MSCs was induced in vitro osteogenesis differentiation,and identified by alizarin red staining. After 3,7,11 and 15 days,h UC-MSCs were individually transplanted into five groups: group A,B,C,D,E,and group F served as the control group with injection of saline solution only. Specimens were obtained individually at 1,2,3 and 4 week after h UC-MSCs transplant. BMP-2 was displayed by immunohistochemical technique and analyzed. The rats' blood routine were also tested. [Result]Many calcium nodes were observed 3 weeks after h UC-MSCs osteogenesis differentiation. The BMP-2 expressions in experimental groups were stronger than that in control group at 1,2,3 and 4 week. There was no significant difference between experimental groups at 1st week and 4th week. But the BMP-2 expressions in group B and group C at 2 week and 3 week were stronger than those in group A,group D and group E. The expression peak appeared at the second week,then went down. As to rats' blood routine,there was no significant difference among all groups. [Conclusion]h UC-MSCs has good immune compatibility. And h UC-MSCs in vitro osteogenesis differentiation can improve the BMP-2 expression in the early stage of fracture healing. The induce effect is the best at the third and seventh days.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2016年第4期362-367,共6页
Orthopedic Journal of China
关键词
人脐带间充质干细胞
体外成骨诱导
骨形态发生蛋白2
大鼠
不全骨折模型
human umbilical cord mesenchymal stem cells(h UC-MSCs)
in vitro osteogenesis differentiation
bone morphogenetic protein 2(BMP-2)
rats
insufficiency fracture model