Kuo Ya Po, Lee Yueh-Lun, Tseng Yu Hong, Su Ching-Hua, Burnouf Thierry, Su Chen Yao
Institute of Oral Biology, National Yang-Ming University, Taipei, Taiwan.
Biologicals. 2010 Jan;38(1):120-7. doi: 10.1016/j.biologicals.2009.07.004. Epub 2009 Aug 31.
Platelet gels (PG) are new topical single-donor blood products which are attracting great interest in regenerative medicine. They are obtained by mixing a platelet-rich plasma fraction with thrombin to generate a fibrin gel enriched in platelet growth factors (GF). The type of thrombin preparation may affect PG reproducibility. We have determined the impact of 14.6% (v/v) ethanol-stabilized thrombin (EHT) on the release of GF by platelets. Various ratios of EHT and platelet concentrates were mixed to obtain from 2.43 to 7.96% ethanol concentration. Platelet-derived growth factor-AB (PDGF-AB), transforming growth factor-beta1 (TGF-beta1), vascular endothelium growth factor (VEGF), epidermal growth factor (EGF), and insulin-like growth factor-1 (IGF-1) were assessed at 5, 120, and 300 min after PG formation. Protein profiles of thrombin and PG releasates were analyzed by SDS-PAGE. The amount of PDGF-AB, TGF-beta1, and VEGF released per platelet decreased significantly (p<0.05) with increasing ethanol concentrations but, however, not that of EGF. IGF-1 content was stable, consistent with its presence mostly in plasma. SDS-PAGE indicated that ethanol did not affect fibrin formation. In conclusion, ethanol has a significant impact on the amount of GF released by platelets and should be strictly controlled to standardize PG and optimize clinical benefits.
血小板凝胶（PG）是新型的局部单供体血液制品，在再生医学领域引起了广泛关注。它们通过将富含血小板的血浆组分与凝血酶混合以生成富含血小板生长因子（GF）的纤维蛋白凝胶而获得。凝血酶制剂的类型可能会影响PG的可重复性。我们已经确定了14.6%（v/v）乙醇稳定化凝血酶（EHT）对血小板释放GF的影响。将不同比例的EHT与血小板浓缩物混合，以获得2.43%至7.96%的乙醇浓度。在PG形成后的5、120和300分钟时，对血小板衍生生长因子-AB（PDGF-AB）、转化生长因子-β1（TGF-β1）、血管内皮生长因子（VEGF）、表皮生长因子（EGF）和胰岛素样生长因子-1（IGF-1）进行了评估。通过SDS-PAGE分析了凝血酶和PG释放物的蛋白质谱。随着乙醇浓度的增加，每个血小板释放的PDGF-AB、TGF-β1和VEGF的量显著减少（p<0.05），然而，EGF的量没有减少。IGF-1含量稳定，与其主要存在于血浆中一致。SDS-PAGE表明乙醇不影响纤维蛋白的形成。总之，乙醇对血小板释放GF的量有显著影响，应严格控制以规范PG并优化临床疗效。
