富血小板血浆对骨髓间充质干细胞向成骨细胞定向分化的影响

Effect of platelet-rich plasma on rabbit bone mesenchymal stem cells differentiation into osteoblasts

目的观察富血小板血浆(platelet-rich plasma,PRP)对兔源骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)定向分化为成骨细胞的作用及机制。方法离心法分离兔骨髓后,体外培养BMSCs,体积分数5%和10%的PRP与BMSCs共培养,观察BMSCs诱导分化过程中茜素红染色阳性率,应用酶标仪检测碱性磷酸酶(alkaliphosphatase,ALP)活性,应用ELISA法检测BMSCs诱导分化过程中骨钙素(osteocalcin,OCN)及结缔组织生长因子(connective tissue growth factor,CTGF)的含量,应用real time-PCR检测成骨相关基因CTGF、ALP、成骨细胞核心结合因子α1(core-binding factor alpha 1,Cbf-α1)、兔Ⅰ型胶原(Col-Ⅰ)和OCN的表达。结果 BMSCs诱导分化后细胞茜素红染色阳性率、ALP活性、OCN及CTGF含量升高,CTGF、ALP、Cbf-α1、Col-Ⅰ和OCN的mRNA表达增加。结论 PRP可促进BMSCs向成骨细胞分化,可能与PRP促进BMSCs中CTGF、ALP、Cbf-α1、Col-Ⅰ和OCN表达有关。

Objective To investigate the effect of platelet-rich plasma（PRP） on rabbit bone mesenchymal stem cells （BMSCs） differentiation into osteoblasts.Methods After BMSCs cultured with 5％ and 10％ PRP in vitro,the positive rate of alizarin red staining for calcium nodules was observed,alkaliphosphatase（ALP） activity was detected by ELIASA,the content of connective tissue growth factor（CTGF） and osteocalcin （OCN） was detected by ELISA,the mRNA expression of CTGF,ALP,core-binding factor alpha 1 （Cbfα1）,Col-Ⅰ and OCN was detected by RT-PCR.Results PRP significantly up-regulated the positive rate of alizarin red staining,ALP activity,CTGF and OCN content,and the mRNA expression of CTGF,ALP,Cbf-α1,Col-Ⅰ and OCN、The up-regulation in 10％PRP group was more significant than in 5％ PRP group.Conclusion PRP could promote BMSCs differentiation into osteoblasts by up-regulating the expression of CTGF,ALP,Cbf-α1,Col-Ⅰ and OCN.