富血小板血浆对人牙髓干细胞／内皮细胞共培养体外成牙本质分化的影响

摘要

AIM:To explore the effects of PRP and the interactions of human dental pulp stem cells (hDP-SCs)with endothelial progenitor cells (EPCs)in the differentiation of hDPSCs into ontoblasts in vitro.METHODS:hDPSCs and EPCs were cultured in vitro and were divided into 6 groups:EPCs+10%FBS,EPCs+10%PRP,hDP-SCs+10%FBS,hDPSCs+10%PRP,hDPSCs+EPCs+10%FBS and hDPSCs+EPCs+10%PRP.The proliferation activity of the cells was detected by MTT assay.the mRNA expression of ALP,DMP-1 and DSPP was examined by qRT-PCR.RESULTS:The proliferation ability of the cells in the cultures with PRP was higher than that with FBS af-ter 5 or 9 days of culture(P<0.05).After 7 or 14 days of culture the expression of ALP,DMP-1 and DSPP in the cultures with PRP was higher than that with FBS (P<0.05)and was higher in co-cultures than in respective cultures (P<0.05).CONCLUSION:PRP can promote the proliferation of hDPSCs and can the promote the differentia of the cells into ontoblasts,especially in co-culture.%目的：探讨人牙髓干细胞（hDPSCs）与内皮祖细胞（EPCs）在体外成牙本质分化中的相互作用以及富血小板血浆（PRP）对其增殖和矿化的影响。方法：体外分离培养hDPSCs和EPCs，并将两者单独或共同接种于含PRP或FBS的DMEM中进行培养，分别于培养1、3、5、7、9 d，MTT法检测各组细胞的增殖活性；然后再按上述相同方法接种细胞并进行矿化诱导，分别于诱导后7 d和14 d，qRT-PCR检测各组细胞的碱性磷酸酶（ALP）、牙本质基质蛋白1（DMP-1）及牙本质涎磷蛋白（DSPP）mRNA的表达水平。结果：无论是单种细胞培养还是两种细胞共同培养，含 PRP各组细胞的增殖活性均明显高于含FBS 各组（P ＜0．05）；ALP、DMP-1、DSPP mRNA的表达水平也均为含PRP组明显高于含FBS组（P＜0．05）；而且在相同培养条件下，两种细胞共同培养组的各基因表达水平均明显高于各细胞单独培养组（P＜0．05）。结论：PRP有维持和促进 hDPSCs、EPCs增殖的能力；并可促进hDPSCs向成牙本质分化，两细胞共同培养时其促进作用更明显。