[The effects of different activators on the release curve of human platelet-rich plasma].

Objective: To compare and analyze the effects of different activators on the release curve of TGF-β(1) and PDGF-AB in platelet rich plasma(PRP). Methods: A total of 36 ml peripheral venous blood was obtained from 10 healthy adult volunteers, and the PRP was made by secondary centrifugation. The platelet activator was made by bovine thrombin 1 000 U in 1 ml 10% calcium chloride solution. The Thrombin-PRP group was made by PRP and the activator in a ratio of 10∶1.The Calcium chloride-PRP group was made in a ratio of 10∶1 by PRP and 10% calcium chloride solution instead. The fresh whole blood(whole blood group) and inactived PRP(PRP group) were used as the control groups. The 4 groups were incubated in warm water of 37 ℃ for 0, 1, 8, 24,72 and 168 h. A quantitative sandwich enzyme-linked immunosorbent assays(ELISA) was used to examine the amount of TGF-β(1) and PDGF-AB in different time points of each group. The release curves of TGF-β(1) and PDGF-AB were based on afore-mentioned data, and then comparisons of the release curves of TGF-β(1) and PDGF-AB in different groups were performed by repeated measurement variance analysis. Results: (1)The levels of TGF-β(1) and PDGF-AB in the whole blood group and the PRP group continued to increase within 168 h. PRP immediately formed into a gel after mixture with thrombin combined and calcium chloride, and the concentrations of TGF-β(1) and PDGF-AB reached the peak in 1 h after activation; increased from (42±21)ng/ml and (77±18)ng/ml to (84±21)ng/ml and (124±35)ng/ml, respectively, and then decreased gradually. The release curve was direct and rapid. The PRP became a gel state in approximate 1 h after mixture with calcium chloride, and the concentrations of TGF-β(1) and PDGF-AB were slowly rising and remained high at 168 h. (2)The AUC(0-168h) of TGF-β(1) and PDGF-AB in the PRP group was higher than that in the whole blood group (all P<0.05) , and the AUC(0-168h) of TGF-β(1) in the Calcium chloride-PRP group was higher than that in the Thrombin-PRP group(Z=-2.26, P<0.05).However, there was no significant difference in the AUC(0-168h) of PDGF-AB between the Calcium chloride-PRP group and the Thrombin-PRP group(Z=-1.512, P=0.131). Conclusion: Using calcium chloride as activator can get a higher release concentration of TGF-β(1) and PDGF-AB and a longer release time, with the largest area under the curve.目的： 对比分析不同激活剂对富血小板血浆（PRP）中转化生长因子-β(1)（TGF-β(1)）及血小板源性生长因子（PDGF-AB）释放曲线的影响。 方法： 抽取10名健康成年志愿者外周静脉全血36 ml，采用二次离心法自制PRP；按1 000 U凝血酶溶于1 ml 10%氯化钙溶液制备血小板激活剂。按10∶1的容量比将PRP与激活剂混合（凝血酶-PRP组)；按10∶1的容量比将PRP与10%氯化钙混合（氯化钙-PRP组)；分别以新鲜全血（全血组）以及未加血小板激活剂的PRP（PRP组）为对照组，将上述4组在37 ℃温水中孵育0、1、8、24、72和168 h，采用ELISA法测定各组不同时间点的TGF-β(1)和PDGF-AB浓度，绘制TGF-β(1)和PDGF-AB的释放曲线，并采用重复测量方差分析比较不同组别中TGF-β(1)和PDGF-AB的释放曲线。 结果： （1）全血组与PRP组中TGF-β(1)和PDGF-AB在168 h内随着时间的推移而持续增高。PRP在与凝血酶混合后，立即形成凝胶状，TGF-β(1)和PDGF-AB均立即明显升高，在激活后1 h达到高峰，TGF-β(1)和PDGF-AB分别由（42±21）和（77±18）μg/L增至高峰（84±21）和（124±35）μg/L，随后逐渐下降，释放曲线直接而快速；而PRP与氯化钙混合后，约1 h才形成凝胶状，TGF-β(1)和PDGF-AB均缓慢持续增高，可在168 h内一直保持较高水平。（2）PRP组中TGF-β(1)和PDGF-AB的AUC(0-168h)均高于全血组（均P<0.05），氯化钙-PRP组中TGF-β(1)的AUC(0-168h)高于凝血酶-PRP组（Z=-2.26，P<0.05），但氯化钙-PRP组与凝血酶-PRP组中PDGF-AB的AUC(0-168h)比较差异无统计学意义（Z=-1.512，P=0.131）。 结论： 使用氯化钙作为激活剂时富血小板血浆中TGF-β(1)和PDGF-AB释放浓度较高，释放时间较长，曲线下面积最大。.