富血小板血浆与洗涤血小板对人牙髓细胞矿化的作用

Effect of platelet-rich plasma and washed platelet on the mineralization of human dental pulp cells

背景:此前课题组的研究表明富血小板血浆和洗涤血小板在一定浓度范围内均可促进人牙髓细胞的增殖。目的:进一步观察不同浓度富血小板血浆和洗涤血小板对人牙髓细胞矿化的作用效果。方法:实验使用健康志愿者正畸拔除牙齿内牙髓培养的4～6代牙髓细胞。将由该志愿者采集的静脉血制备富血小板血浆和洗涤血小板作用于牙髓细胞,使用碱性磷酸酶及蛋白定量试剂盒测定矿化诱导7d后牙髓细胞的碱性磷酸酶活性,并通过茜素红染色观测牙髓细胞经矿化诱导10d及20d后的矿化结节形成情况。结果与结论:10%～30%的洗涤血小板与富血小板血浆均明显提高了牙髓细胞碱性磷酸酶活性,其中,以20%浓度尤为明显;10%～30%的洗涤血小板与富血小板血浆均明显促进了矿化诱导后10d的牙髓细胞矿化结节形成,其中,10%浓度在矿化诱导后20d促进牙髓细胞形成的矿化结节最大。但相同浓度的洗涤血小板与富血小板血浆之间在作用效果上没有明显差异。

BACKGROUND：Previous studies of our research group have indicated that both washed platelet and platelet-rich plasma promote the proliferation of human dental pulp cells in some concentration range.OBJECTIVE：To further investigate the effect of washed platelet and platelet-rich plasma at different concentrations on the mineralization of human dental pulp cells.METHODS：Human dental pulp cells from the healthy extracted teeth donated by patients under orthodontic treatment were cultured and passaged for 4-6 passages.Platelet-rich plasma and washed platelet were manufactured from venous blood of the same donor.The activity of alkaline phosphatase in dental pulp cells was determined using Alkaline Phosphatase Reagent Kit and Protein Quantification Reagent Kit on the 7th day after mineralization induction.The formation of mineralized nodules in dental pulp cells were observed by alizarin red staining on the 10th and 20th days after mineralization induction.RESULTS AND CONCLUSION：Both washed platelet and platelet-rich plasma at the concentration of 10%-30% evidently promoted the alkaline phosphatase activity in dental pulp cells.The most effective concentration to promote the alkaline phosphatase activity was 20%.Both washed platelet and platelet-rich plasma at the concentration of 10%-30% evidently promoted the formation of mineralized nodules in dental pulp cells on the 10th day after mineralization induction.The mineralized nodules in the 10% concentration group were the largest on the 20th day after mineralization induction.There was no difference between the washed platelet and platelet-rich plasma at the same concentration.