富血小板血浆凝胶对家兔骨髓间充质干细胞增殖与成骨活性的影响  

Effects of platelet-rich plasma gel on the proliferation and osteogenic activity of rabbit bone marrow mesenchymal stem cells

作　　者：张林朴[1,2,3] 代晓华[2,3] 王冠华[2,3] 李燕妮[2,3] 连小丽[2,3] 姚晖[2,3] 

机构地区：[1]天津医科大学口腔医学院,天津市300070 [2]天津市口腔医院 [3]南开大学附属口腔医院,天津市300041

出　　处：《中国组织工程研究》2012年第23期4199-4203,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:富血小板血浆含有多种骨组织修复有关的生长刺激因子,并且在凝聚后可以形成纤维蛋白网络支架利于细胞的黏附而促进骨组织再生。目的:体外评价家兔骨髓间充质干细胞在富血小板血浆凝胶中的增殖与成骨活性。方法:分离培养5d龄新生大耳白兔骨髓间充质干细胞;提取成年大耳白兔外周静脉血的富血小板血浆。设置富血小板血浆复合骨髓间充质干细胞(复合细胞组)、复合α-MEM培养液组及单纯骨髓间充质干细胞组进行观察。结果与结论:各组间的乳酸脱氢酶活性比较差异有显著性意义(P<0.05),复合细胞组骨髓间充质干细胞增殖明显优于其他组,其凝胶完整、均匀、半透明,细胞不规则多角形散在分布,有较多长突触伸展在凝胶中,在凝胶中散在分布黄绿色荧光亮点和结节,显示有钙化结节形成。数量和大小第3周比第2周稍有增多和增大,而与α-MEM培养液复合未见荧光显色。说明在富血小板血浆凝胶复合骨髓间充质干细胞后,明显促进了骨髓间充质干细胞的增殖与成骨活性。BACKGROUND： Platelet-rich plasma （PRP） contains several kinds of growth factors related to bone repair, and can form fibrin meshwork scaffold after agglomerated by activation, which is beneficial to cells adhesion and promotes bone regeneration. OBJECTIVE： To evaluate the proliferation and osteogenic activity of rabbit bone marrow mesenchymal stem cells （rBMSCs） in platelet-rich plasma gel in vitro. METHODS： rBMSCs were isolated from bone marrow of limb long bone of 5-day neonatal rabbits by adhesion culture. PRP was obtained from adult rabbit peripheral venous blood. Experiment samples were divided into three groups： PRP plus rBMSCs （PRP/rBMSCs） group, PRP plus a-MEM medium （PRP/a-MEM） group and rBMSCs group. RESULTS AND CONCLUSION： The difference of lactate dehydrogenase activity among three groups was significant （P 〈 0.05）, and the cell proliferation in PRP/rBMSCs group was obviously better than that in the other two groups. The PRP/rBMSCs gel was complete, uniform and semitransparent. It had many cells which were anomalistic polygon with long synapse distributing in gel. Tetracycline labeled flavo-green fluorescence brightened dots and nodes were discovered in PRP/rBMSCs gel after 2 and 3 weeks of culture using fluorescence microscope. Calcify nodes were formed in the PRP/rBMSCs gel. The number and size of fluorescence brightened dots and nodes founded in the third week after culture were a little more and bigger than those found in the second week. There were no fluorescence brightened dots and nodes in PRP/a-MEM gel. It indicates that rBMSCs combined with PRP gel can promote the proliferation and osteogenic activity of rBMSCs.

关 键 词：富血小板血浆 骨髓间充质干细胞 细胞增殖 钙化结节 乳酸脱氢酶 干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]