PRP联合盐酸川芎嗪对BMSCs体外增殖及向成骨细胞分化的影响

Impacts of PRP and ligustrazine hydrochloride on BMSCs in vitro prolif-eration and definitive differentiation to osteoblast

目的通过将不同比例的富血小板血浆(Platelet-rich plasma,PRP)与盐酸川芎嗪联合用于人骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs),观察不同比例对其体外增殖及其向成骨细胞分化的影响。方法选取我院住院并签订知情同意书的患者为研究对象,抽取已签订知情同意书的患者静脉血50m L,分别二次离心法制备PRP,进行血小板计数分析。根据骨髓间充质干细胞培养是否加入药物为分组依据,分成A、B、C、D四组,A组为对照组,培养液中不加入药物,其余三组分别将体积分数为15%、30%、50%的PRP与80%盐酸川芎嗪按照1∶1.5混合加入骨髓间充质干细胞的培养液中,检测细胞的增殖状况,细胞达到80%融合,消化收集细胞;计数确定各组细胞培养情况,确定加入PRP和盐酸川芎嗪组对细胞培养的影响。对加入PRP和盐酸川芎嗪组培养的细胞进行成骨诱导,确定其成骨分化特性。结果各组PRP+盐酸川芎嗪促进MSCs增殖优于对照组。达到80%融合时间更短。随着PRP浓度增加细胞增殖效率显著增加。结论 PRP与盐酸川芎嗪对BMSCs的体外增殖有促进作用,与PRP体积分数和作用时间呈正相关关系。

Objective To apply the combined administration of platelet-rich plasma（PRP）and 1i-gustrazine hydrochloride on human bone marrow mesenchymal stem cell（BMSCs） at different ratios and observe the impacts on the in vitro proliferation and definitive differentiation to osteoblast. Methods Select hospital and signed informed consent in our hospital patients as the research object, the extraction have been signed informed consent of vein blood of patients with 50 m L,two PRP were prepared by centrifugal casting, the platelet count analysis. According to the bone marrow mesenchymal stem cells cultured on whether or not to join the drug is the grouping basis, divided into A, B, C, D four groups, group A was the control group, the drug does not join in culture liquid, the other three groups respectively, the volume fraction of 15%, 30%, 50% PRP and 80% of ligustrazine hydrochloride in accordance with 1： 1.5 mix bone marrow mesenchymal stem cells were cultured in the media, the detection of cell proliferation, cell reached 80% confluence, digestive cells were collected; count was determined in cell culture conditions, to determine the influence of joining PRP and ligustrazine hydrochloride group on cell culture. To join the PRP and ligustrazine hydrochloride group cultured cells of osteogenic induction, determine the osteogenic differentiation. Results PRP＋ligustrazine hydrochloride promoted BMSCs proliferation in each group, which was superior to the control group and 80% of infusion was obtained in very short time. With the increase of PRP concentration, the efficiency of cell proliferation was enhanced significantly. Conclusion PRP and ligustrazine hydrochloride promote the in vitro proliferation of human BMSCs, which presents the positive correlation with PRP volume fraction and effect time.