富血小板血浆促进骨髓间充质干细胞对大鼠骨缺损的修复作用

Effect of the promotion of platelet-rich plasma in the repair of bone defects in rats with bone marrow mesenchymal stem cells

目的探讨富血小板血浆(platelet-rich plasma, PRP)对大鼠骨缺损的修复作用。方法提取大鼠骨髓间充质干细胞(bone marrow stromal cells, BMSCs),获得血小板浓度为2×10~5/L、2×10~6/L和2×10~7/L的PRP,分别将不同浓度的PRP与BMSCs共培养,通过MTT实验观察PRP对BMSCs细胞增殖的影响,ELISA实验检测PRP对BMSCs中碱性磷酸酶(alkaline phosphatase,ALP)的作用。构建大鼠股骨节段缺损模型,分别使用BMSCs和PRP(血小板2×10~5/L)混合物;BMSCs和PRP(血小板2×10~6/L)混合物;BMSCs和PRP(血小板2×10~6/L)来填补模型中的缺陷部位。8周后处死大鼠,评估缺损修复情况。通过Western Blot和Real-Time PCR检测断端骨组织内BMP-2和Collagen I含量,免疫组织化学染色检测组织内BMP-2含量。结果不同浓度的PRP对BMSCs的增殖和ALP的表达情况均有促进作用,且随血小板浓度的增加而加剧。PRP的添加能够促进大鼠骨缺损模型中的骨再生作用,促进BMP-2和Collagen I的表达,其促进作用具有剂量依赖性。BMSCs单独处理组能够在一定程度上促进骨缺损的修复,但作用较慢。PRP单独应用对骨缺损的修复作用较小。结论 PRP具有促进BMSCs对大鼠骨缺损的修复作用。

Objective To explore the effect of the promotion of platelet-rich plasma(PRP)in the repair of bone defects in rats with bone marrow mesenchymal stem cells.Methods Bone marrow stromal cells(BMSCs)were extracted from rat bone marrow and PRP with platelet concentrations of 2×10~5/L,2×10~6/L and 2×10~7/L was obtained.Different concentrations of PRP were co-cultured with BMSCs.The effect of PRP on the proliferation cof BMSCs was observed by MTT assay.The effect of PRP on alkaline phosphatase(ALP)in BMSCs was detected by ELISA assay.After construction of a rat femoral segmental defect model,a BMSCs and PRP(platelet was 2×10~5/L,2×10~6/L and 2×10~7/L)mixture was injected into rat femur defects.All rats were euthanized after 8 weeks and the repair of the defects was evaluated.The content of BMP-2 and Collagen I in the broken femur was detected by Western Blot and Real-Time PCR.The content of BMP-2 in tissues was detected by immunohistochemical staining.Results Both proliferation of BMSCs and expression of ALP can be promoted by different concentrations of PRP.The higher the platelet concentration,the more obvious the effect.PRP can promote the regeneration of bone and the expression of BMP-2 and Collagen I in a rat femoral segmental defect model in a dose-dependent manner.The action of BMSCs alone was relatively slow,and the effect of PRP alone was unremarkable on the repair of the femur defects.Conclusion PRP can promote the repair of BMSCs in femur defects in rats.