复合富血小板血浆成骨微环境对兔骨髓间充质干细胞增殖分化的影响  

Effect of platelet-rich plasma with bone marrow tissue fluid on proliferation and osteogenic differentiation of rabbit bone marrow messenchymal stem cells

作　　者：程文俊[1] 左伟[1] 肖志宏[1] 李涛[1] 黄若昆[1] 严立[1] 王俊文[1] Cheng Wenjun Zuo Wei Xiao Zhihong Li Tao Huang Ruokun Yan Li Wang Junwen(Department of Orthopaedics and Traumatology , Puai Hospital, Tongii Medical College, Huazhong University of Science and Technology, Wuhan 430033, Chin)

机构地区：[1]华中科技大学同济医学院附属普爱医院骨科,武汉430033

出　　处：《中华实验外科杂志》2016年第11期2469-2471,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省研究与开发计划项目（2011BCB028）;湖北省卫生厅青年科技人才项目（QJX2010-46）;武汉市卫生局基金（WX09A05）

摘　　要：目的 观察富血小板血浆（PRP）与骨髓组织液对体外培养的兔骨髓间充质干细胞（BMSCs）增殖和成骨分化的影响,探讨PRP促进骨修复的机制.方法 将兔MSCs分别经复合RPR的骨髓组织液和单纯血清培养,取培养后3代MSCs,分为单纯血清培养（FCS）组和PRP组（PRP骨髓组织液复合物）,采用噻唑蓝（MTT）法检测第2、4、6、8天时细胞增殖活性;测定培养后第3、7、10天碱性磷酸酶（ALP）活性;ALP染色、矿化结节染色检测细胞成骨分化活性.结果 MTT法显示PRP组2、4、6、8d的平均吸光度（A）值分别为0.193 ±0.041、0.476±0.050、0.786±0.122、1.216 ±0.139;对照组为0.169±0.054、0.448±0.061、0.835±0.104、1.165±0.162;各个时间点PRP组与对照组相比差异无统计学意义（P＞0.05）.ALP活性测定PRP组3、7、10d分别为2.233±0.276、4.528 ±0.516、3.922±0.603;对照组为1.648±0.340、2.457±0.541、2.140±0.269;各个时间点PRP组均高于对照组,差异有统计学意义（P＜0.01）.PRP组ALP阳性细胞数为（54.71±11.48）％、钙结节为（110.5±20.9）个/孔,对照组分别为（6.25±2.41）％、（9.2±3.7）个/孔;与对照组相比,PRP组ALP阳性细胞数增多（P＜0.01）,矿盐沉积增多（P＜0.01）,差异有统计学意义.结论 复合PRP骨髓组织液诱导的MSCs能恢复正常的细胞增殖速率,能诱导MSCs向成骨细胞分化,这有利于PRP促进骨组织修复.Objective To study the action mechanism of platelet-rich plasma （PRP） in bone tissue repairing by investigating the effect of PRP with bone marrow tissue fluid on proliferation and osteogenic differentiation of rabbit bone marrow messenchymal stem cells （MSCs）.Methods MSCs treated by PRP with bone marrow tissue or untreated were divided into two groups：fetal calf serum group （FSC group）,PRP group （MSCs treated by PRP and bone marrow tissue fluid）.The methyl thiazol tetrazolium （MTT） assay was used to evaluate cell proliferation at the day 2,4,6,and 8.Alkaline phosphate （ALP） activity was measured at day 3,7,and 10.ALP and calcium staining were used to evaluate the effect of PRP on osteogenic differatiation of MSCs.Results The MTT assay showed that at 2,4,6 and 8 days the mean absorbance values were 0.193 ± 0.041,0.476 ± 0.050,0.786 ± 0.122,1.216 ± 0.139 in PRP group,while 0.169 ± 0.054,0.448 ± 0.061,0.835 ± 0.104,1.165 ± 0.162 in the FCS group.There was no signifigant difference between PRP group and FSC group regarding their effects on proliferationon day 2,4,6,8 （P 〉 0.05）.The absorbance value of ALP activity tests were 2.233 ± 0.276,4.528 ± 0.516,3.922 ±0.603 in PRP group,while 1.648 ±0.340,2.457 ±0.541,2.140 ±0.269 in FCS group at 3,7,10 days.Compared to FCS group,PRP group promoted ALP secret at 3,7,10 days（P 〈0.01）.The number of ALP-positive cells proportion and calcium nodes was （54.71 ± 11.48） % and （110.5 ± 20.9）/hole respectively in PRP group,and （6.25 ± 2.41）%,（9.2 ± 3.7）/hole in FCS group.Compared to FCS group,PRP group increased the number of ALP-positive cells （P 〈 0.01） and calcium nodes （P 〈0.01）.Conclusion MSCs treated by PRP with bone marrow tissue fluid can restore to the normal proliferation rate.PRP with bone marrow tissue fluid can promote osteogenic differentiation of MSCs.

关 键 词：骨髓间充质干细胞 富含血小板血浆 增殖 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]