富血小板血浆与地塞米松对人骨髓基质干细胞成骨分化作用的对比研究

目的 通过系统评价富血小板血浆(PRP)与地塞米松(DEX)对人骨髓基质干细胞(BMSCs)成骨分化的影响,为PRP临床骨组织修复应用提供更为可靠的实验依据.方法 将体外培养的BMSCs分为单纯血清培养组(FCS组)、PRP诱导组和DEX组,通过相差显微镜观察碱性磷酸酶(ALP)染色、钙盐沉积染色,RT-PCR法检测碱性磷酸酶(ALP)、骨钙素(OC)、Ⅰ型胶原(Coll-Ⅰ)、骨连接蛋白(ON)、中心结合因子(Cbfα1)mRNA表达系统评价PRP的成骨分化能力. 结果 PRP抑制了BMSCs向三角形、多角形细胞转变,DEX则诱导BMSCs向三角形、多角形细胞转变;PRP抑制了ALP分泌,钙盐沉积;DEX增加了ALP分泌,促进钙化结节形成.与FCS组相比,DEX促进了ALP、OCmRNA表达,PRP抑制了ALP、OC mRNA表达;PRP、DEX对Coll-Ⅰ、ON、Cbfα1 mRNA表达均无影响.结论 在本实验条件下,PRP对人BMSCs体外成骨分化的直接作用是抑制效应;在体外PRP并不能代替DEX作为人BMSCs成骨分化的诱导因子.

Platelet-rich plasma versus dexamethasone in inhibiting effect on osteogenic differentiation of human bone marrow mesenchymal stem cells

Objective To compare the effects of platelet-rieh plasma (PBP) and dexamethasone (DEX) on osteogenic differentiation of human bone marrow mesenchymal stromal cells (BMSCs). Methods BMSCs cultured in vitro were divided to 3 groups: fetal calf serum group (FCS group), PRP group (human BMSCs treated by PRP), and DEX group (human BMSCs treated by DEX) . Inverted phase contrast mi-croscopy, alkaline phosphate (ALP) staining, calcium staining and RT-PCR were used to determine gene expressions of ALP, osteocalcin (OC), collagen type Ⅰ (Coll-Ⅰ), osteonectin (ON), and core binding factor alpha 1 (Cbfα1) . The effects of PRP on osteogenic differentiation of human BMSCs were evaluated and compared with those of DEX. Results Most of the BMSCs treated by DEX were triangle or polygon in form while those treated by PRP were fusiform. At day 7, compared with FCS group, the number of ALP-positive cells in PRP group decreased (P < 0.05), but increased in DEX group (P < 0.05). At day 19, there was a significant difference in mineral deposition between PRP and FCS groups, with few calcium nodes in the PRP group (P < 0.05) . Compared with FCS group, treatment of Human BMSCs with DEX markedly increased mineral deposition, leading to much more calcium nodes (P < 0.05). DEX resulted in an earlier expression of OC at day 7 compared with FCS, and increased OC mRNA expressions at other culture time points. At the same time, DEX up-regulated ALP mRNA expression. Compared with FCS group, PRP decreased OC and ALP mRNA expressions. At different time points, the expressions of Cbfα1, ON, and Coll-Ⅰ mRNA showed no significant changes in the 3 groups. Conclusion PRP may have a direct effect on inhibiting osteogenic differentiation of human BMSCs, and can not be a substitute of DEX as an agent of osteogenic differentiation in vitro.