摘要
目的：探讨骨形态发生蛋白质-4（BMP-4）基因转染骨髓间充质干细胞（BMSCs）复合聚乳酸羟基乙酸共聚物（PLGA）修复兔全层关节软骨缺损的效果。方法：取兔骨髓间充质干细胞，分离培养后，免疫组织化学鉴定BMSCs。采用电转法将pcDNA3-1-BMP-4质粒导入BMSCs。转染成功后培养备用。制备直径为4mm的双层PLGA支架。新西兰大白兔随机分成空白组、PLGA组、PLGA/BMSCs组、PLGA/BMP-4/BMSCs组4组，将双层PLGA支架置入兔双膝关节股骨内侧髁软骨缺损中。观察并记录术后动物膝关节活动情况。第8、16周时取膝关节置入组织行H-E染色，在扫描电镜下观察置入的PLGA新生组织。采用RT-PCR法定量检测4组修复软骨缺损处新生的软骨Ⅱ型胶原、SOX9、蛋白聚糖基因的表达水平。结果：术后各组动物膝关节活动正常，未见炎症反应。第8、16周时PLGA/BMP4/BMSCs组兔膝关节损伤修复优于其他3组。H-E染色结果可见PLGA/BMP4/BMSCs组膝关节损伤修复效果优于其他3组。RT-PCR结果显示PLGA/BMP4/BMSCs组兔置入的PLGA新生的软骨中Ⅱ型胶原、SOX-9、蛋白聚糖基因的表达水平显著高于其他各组。结论：BMP-4转染BMSCs后能够促进骨髓间充质干细胞分化成软骨细胞，双层PLGA支架置入有利于兔膝关节软骨缺损区的修复。
Objective： To investigate the effect of repairing for knee articular cartilage defects by double layered PLGA laden with BMSCs-BMP-4. Methods： Rabbit BMSCs were isolated and cultured, and immunohistoehemical method was used to identify the BMSCs. The plasmid pcDNA 3.1-BMP-4 was successfully introduced into BMSCs by electroporation. A double layer PLGA scaffold with a diameter of 4 mm was set and implanted in the knee joint. Forty New Zealand white rabbits were randomly divided into four groups （n = 10） ： blank group, PLGA group, BMSCs PLGA/ group, PLGA/BMP-4/BMSCs group. The activities of knee joint in rabbits after operation were observed and recorded. H-E staining was performed to observe knee joint structure at eighth week and sixteenth week. Implanted PLGA was observed under scanning electron microscope. RT-PCR method was used to detect the expression of collagen type U , SOX-9 and aggrecan in the new cartilage in four groups. Results： There was no inflammatory reaction in the knee joint after operation. H-E staining results showed that the repair in the PLGA/BMP-4/BMSCs group was better than that in the other 3 groups. RT-PCR results showed that the expression of collagen type 11 , SOX-9 and aggrecan in the PLGA group was significantly higher than that in other groups. Conclusion： BMP-4 can promote the differentiation of bone marrow mesenchymal stem cells into ehondrocytes, and double PLGA stent implantation is beneficial to the cartilage defect repair of rabbit knee joint.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2016年第6期667-672,共6页
Chinese Journal of Anatomy
