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目的分析人血小板及其洗涤后获得的各组分补体灭活前后对小鼠成纤维细胞(L929)和人成纤维细胞(HDP)的增殖作用。方法将所制备的新鲜人富血小板血浆(PRP)3份及血小板浓缩液(PC)5份(50 mL/份),分别用20 mL 0.9%生理盐水洗涤2次,重悬以获得贫血小板血浆(PPP)、生理盐水上清、洗涤血小板(WP)3个组分,同未经处理的PRP及PC一道,分别冻融留取对照样品后作补体灭活处理,ELASA方法测试补体灭活前后血小板源性生长因子(PDGF-AB)、转化生长因子(TGF-β1)、血管内皮生长因子(VEGF)、表皮生长因子(EGF)含量,CCK-8试剂盒测定补体灭活前后对L929/HDP细胞的增殖作用。结果 PRP及洗涤各组分补体灭活前后PDGF-AB、VEGF及EGF含量比较,P>0.05,PC及各组分补体灭活前后PDGF-AB、VEGF、EG及TGF-β1含量比较均无明显差异(P>0.05)。PRP及WP促L929细胞增殖比较:PRP为0.68±0.12 vs 2.13±0.18(P<0.05),WP为0.69±0.12 vs 0.66±0.13(P>0.05);PC及WP促L929细胞增殖比较:PC为0.42±0.05 vs 1.94±0.19(P<0.05),WP为1.69±0.13 vs 1.93±0.19(P>0.05)。补体灭活前后,PRP及WP促HDP细胞增殖:PRP为1.36±0.09 vs1.30±0.11(P>0.05),WP为1.26±0.04 vs 1.33±0.10(P>0.05);PC洗涤组分促细胞增殖:PC为1.32±0.07 vs1.28±0.09(P>0.05),WP为1.39±0.13 vs 1.44±0.13(P>0.05)。结论补体灭活对PRP与PC及洗涤获得的各组分PGFs含量影响较小,PGFs含量与促L929/HDP细胞的增殖作用存在非线性正相关关系,PGFs促细胞增殖可能存在种间差异。
Objective To investigate the proliferation of mouse fibroblasts (L929) and human fibroblasts (HDP) before and after inactivation of human platelets and their components after washing. Methods The prepared fresh human platelet-rich plasma (PRP) 3 and platelet concentrate (PC) 5 copies (50 mL / copies) were washed twice with 20 mL of 0.9% saline and resuspended to obtain platelet-poor plasma (PPP), saline supernatant and washing platelet (WP). The untreated PRP and PC were respectively frozen and thawed for control inactivation. The ELASA method was used to test the effect of complement inactivation The levels of platelet-derived growth factor (PDGF-AB), transforming growth factor (TGF-β1), vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) Cell proliferation. Results The levels of PDGF-AB, VEGF and EGF before and after inactivation of complement components of PRP and washing fractions were compared, P> 0.05. The contents of PDGF-AB, VEGF, EG and TGF- Difference (P> 0.05). PRP and WP promoted the proliferation of L929 cells: PRP was 0.68 ± 0.12 vs 2.13 ± 0.18 (P <0.05), WP was 0.69 ± 0.12 vs 0.66 ± 0.13 (P> 0.05) 0.42 ± 0.05 vs 1.94 ± 0.19 (P <0.05), WP was 1.69 ± 0.13 vs 1.93 ± 0.19 (P> 0.05). PRP and WP promoted the proliferation of HDP cells before and after inactivation of PRP: 1.36 ± 0.09 vs1.30 ± 0.11 (P> 0.05), WP was 1.26 ± 0.04 vs 1.33 ± 0.10 (P> 0.05) Proliferation: PC was 1.32 ± 0.07 vs 1.28 ± 0.09 (P> 0.05). WP was 1.39 ± 0.13 vs 1.44 ± 0.13 (P> 0.05). Conclusions Inactivation of complement has little effect on the content of PGFs in each component of PRP, PC and washing. There is a non-linear positive correlation between the content of PGFs and the proliferation of L929 / HDP cells, and there may be some differences among them.