不同比例激活富血小板血浆对其生长因子浓度及人骨髓基质干细胞增殖的影响

目的 确定激活剂凝血酶与富血小板血浆(PRP)混合的最适比例,并探讨凝血酶与PRP中血小板衍生生长因子.AB(PDGF.AB)和转化生长因子-β(TGF-β)对人骨髓基质干细胞(hBMSCs)增殖的联合作用.方法 按1000 U凝血酶溶于1 mL质量百分比为10%的CaCl2配制激活剂.按PRP与激活剂混合比例为1:1、5:1、10:1、20:1、40:1分为A、B、C、D、E 5组.ELISA法测定各组0、1、8、24、120 h后PRP凝胶中PDGF-AB、TGF-β1的浓度;MTT法评价各组PRP凝胶上清对hBMSCs增殖的影响.结果 PRP与凝血酶混合后,PDGF-AB、TGF-β1浓度立即升高,在激活1 h后达到高峰.B、C、D组PRP凝胶中PDGF-AB、TGF-β1的浓度最高,并明显促进hBMSCs的增殖;而A组则抑制hBMSCs的增殖.结论 PRP对hBMSCs增殖的作用与其所含有PDGF-AB、TGF-β1存在浓度依赖性,而凝血酶可能在一定浓度范尉内主要通过对PRP中生长因子浓度的调控来影响hBMSCs的增殖,但是当凝血酶浓度过高时则抑制hBMSCs的增殖.

Effect of different ratio activation on concentrations of PDGF-AB and TGF-β1 in PRP and prolifera-tion of human marrow-derived mesenchymal stem cells

Objective To explore the best ratio of platelet-rich plasma (PRP) to activator and the synergistic action of thrombin and the growth factors in PRP gel on proliferation of human marrow-derived mesenchymal stem cells (hBMSCs). Methods The activator was made up of 1000 U bovine thrombin and 1 mL 10% calcium chloride solution. PRP was mixed with the activator at the ratios of 1:1, 5:1, 10:1, 20:1, 40:1 respectively in groups A, B, C, D and E. A quantitative sandwich enzyme-linked immunosorbent assay (ELISA) was used for examining the amounts of transforming growth factor-β1 (TGF-β1) and platelet derived growth factnr-AB (PDGF-AB) in PRP gel in each group after incubation for 0,1,8,24, and 120 hours. MTT assay was used to evaluate the effect of PRP gel in each group on hBMSCs proliferation. Results When PRP was mixed with thrombin, concentrations of PDGF-AB and TGF-β1 in PRP gel increased immediately and reached the peak value in 1 hour. The PRP gel in groups B, C, and D contained the highest amounts of PDGF-AB and TGF-β1, and accelerated hBMSCs growth re-markably. The cells proliferation in group A was inhibited. Conclusions The effect of PRP on the hBMSCs proliferation depends on the concentrations of PDGF-AB and TGF-β1 in PRP. Thrombin may influ-ence the hBMSCs proliferation by regulating concentrations of growth factors in PRP to certain extent, but too high a level of thrombin may inhibit hBMSCs growth.