猪富血小板血浆冻干粉细胞因子释放的研究

Analysis of release kinetics of growth factors in porcine platelet - rich plasma lyophilized powder

目的在制备较高血小板浓度的猪富血小板血浆（PRP）基础上，探索PRP冷冻干燥（FD）方法，研究PRP冻干粉中细胞因子释放动力学。方法实验分组：静置PRP组、PRP钙剂激活组（CaPRP）、PRP激活后冻干组（Ca-FDPRP）、PRP冻干后激活组（FD—CaPRP）。所有FDPRP样本室温（22℃）密封保存4周，检测样本转化生长因子-β1（TGF-β1）、血小板来源生长因子-AB（PDGF—AB）和血管内皮生长因子（VEGF）在15min和1h的浓度。结果CaPRP组1h的细胞因子浓度均高于相同时间点的PRP组（p〈0．05），PDGF-AB在FD—CaPRP组[15min：（6137．5±800．3）ng／L；1h：（5659．1±656．4）ng／L]和CaPRP组[15min：（5629．8±580．1）ng／L；1h：（5294．6±679．2）ng／L]组间比较差异无统计学意义（P〉0．05）。VEGF在Ca．FDPRP组[15min：（942．6±178．5）ng／L；1h：（895．8±110．O）ng／L]和CaPRP组[15min：（589．7±272．9）ng／L；1h：（782．5±109．5）ng／L）在相同时间比较差异无统计学意义（P〉0．05）。TGF-β1在Ca-FDPRP组15min的浓度[（14801．5±445．2）ng／L]高于CaPRP组[（7812．6±571．0）ng／L，P〈0．05]。结论PRP可以被钙剂有效激活，PRP冻干粉室温保存4周后仍富含细胞因子，且先激活后冻干的PRP使用较为方便。

Objective On the basis of acquiring enough porcine platelet rich plasma （PRP） with high concentration of platelets, we further seek an appropriate way to preserve PRP by lyophilization and ex- plored the kinetics of growth factors released by PRP lyophilized powder. Methods Four experimental groups were arranged as following： （1） Fresh porcine PRP, （2） Group of PRP activated by calcium chloride （CaCl2 ） （Ca PRP）, （3） Activated PRP followed by freeze drying （Ca- FD PRP）, and （4） Freeze -dried PRP activated by CaCl2 （ FD - Ca PRP）. All FD PRP samples were kept for 4 weeks at room temperature （22 ℃ ）. Concentrations of transforming growth factor - β1 （ TGF - β1 ）, platelet - derived growth factor -AB （PDGF -AB）, and vascular endothelial growth factor （VEGF） were determined at time points of 15 min and 1 h respectively. Results Concentration of all growth factors in Ca PRP at 1 h were signifi- cantly higher than that in the PRP at same time point （P 〈 0. 05）. There was no statistical significance in variance of PDGF - AB concentrations between FD - Ca PRP [ 15 min： （6 137.5 ± 800. 3） ng/L; 1 h： （5659.1± 656.4） ng/L] and Ca PRP [15min： （5629.8± 580.1） ng/L; 1 h： （5294.6± 679. 2） ng/L, P〉0. 05]. Levels of VEGF between Ca - FD PRP [ 15 min： （942. 6 ± 178. 5） ng/L; lh： （895.8± 110.0） ng/L] and Ca PRP [15 min： （589.7± 272.9） ng/L; 1h： （782.5± 109. 5） ng/L] was not significantly different （P〉0. 05）. However, TGF - β1 concentration in Ca - FD PRP[ （ 14 801.5 ±445.2） ng/L] at 15 rain was higher than that in Ca PRP [ （7 812. 6 ± 571.0） ng/L, P 〈0. 05 ]. Conclusion PRP can be activated efficiently by calcium chloride. Lyophilized PRP is still rich in growth factors after four weeks＇ preservation at room temperature, which reveals easy - to - use per- formance and wide possibility for clinical applications.