CD41 Western blotting: a new method to detect platelet adhesion to artificial surfaces used in extracorporeal circulation procedures
CD41 Western blotting: a new method to detect platelet adhesion to artificial surfaces used in extracorporeal circulation procedures
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DOI：
10.1007/s10856-008-3587-y
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发表时间：
2009-01-01
影响因子：
3.7
通讯作者：
Wendel, Hans Peter
中科院分区：
文献类型：
--
作者：
Paul, Angela;Straub, Andreas;Wendel, Hans Peter
心脏搭桥术（CPB）手术与血小板活化和血小板计数减少有关，这是由于血液成分的人工表面活化和非生理性流动模式。如先前的研究所示，医疗器械的涂层可以改善体外循环系统中的血液相容性。在这项研究中，我们展示了一种新的方法来确定血小板粘附18涂层和非涂层膜式氧合器在模拟CPB模型与CD 41蛋白质印迹。在120分钟的再循环阶段测定血小板损失和β-TG（血小板活化标志物）的释放。在运行结束时，冲洗膜式氧合器（带管路系统），并使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹技术分析表面吸附的蛋白质量。与涂层膜式氧合器相比，无涂层器械显示出显著更高的CD 41和纤维蛋白原吸附浓度。这些结果与β-TG释放和血小板损失一致，表明与未涂层组相比，涂层氧合器上的血小板粘附较少。这种新方法可能有助于为所有类型的血液接触装置选择较少的血小板活化材料，以改善促凝性，包括血小板功能。
Cardiopulmonary bypass (CPB) surgery is associated with platelet activation and reduced platelet counts due to artificial surface activation of blood elements and non-physiological flow-patterns. As shown in former studies, coating of medical devices can improve hemocompatibility in extracorporeal circulation systems. In this study, we demonstrate a new method to determine platelet adhesion on 18 coated and non-coated membrane oxygenators in a simulated CPB model with CD41 Western blot. Platelet loss and the release of beta-TG (platelet activation marker) were determined during a 120 min recirculation phase. At the end of the run the membrane oxygenators (with tubing system) were rinsed and the amount of adsorbed proteins on the surface was analyzed by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting technique. Uncoated devices showed significantly higher concentrations of CD41 and of fibrinogen adsorption compared to the coated membrane oxygenators. These results correspond with the release of beta-TG and platelet loss indicating less platelet adhesion on the coated oxygenators compared with the uncoated group. This new method may be useful in choosing less platelet activating materials for all kind of blood contacting devices to improve thrombogenicity including platelet functionality.