Microfluidic assay of antiplatelet agents for inhibition of shear-induced platelet adhesion and activation.
Microfluidic assay of antiplatelet agents for inhibition of shear-induced platelet adhesion and activation.
复制标题
抗血小板药物抑制剪切诱导的血小板粘附和活化的微流体测定。
DOI：
10.1039/d0lc00756k
复制
发表时间：
2021
期刊：
影响因子：
6.1
通讯作者：
Hlady,Vladimir
中科院分区：
文献类型：
--
作者：
Rahman,ShekhMojibur;Hlady,Vladimir
我们已经开发了一种微流体系统，通过具有上游狭窄区和下游蛋白质捕获区的流动通道灌注全血。这种基于血流的系统被用来检测抗血小板药物如何有效地抑制剪切诱导的血小板粘附和下游狭窄区域的活化。微接触打印用于将三种血小板结合蛋白[纤维蛋白原、胶原蛋白或血管性血友病因子（vWf）]中的一种共价附着在下游捕获区表面。含抗血小板剂的全血瞬间暴露于上游高壁剪切速率（4860 s - 1或11 560 s - 1），随后流过下游捕获区，在那里测量血小板粘附性。几种抗血小板药物（乙酰水杨酸、替罗非班、依替巴肽、抗vwf和抗gpib α）对减弱下游粘附的效果进行了评估。在抗体阻断vWf或GPIbα后，流式细胞术还使用两种激活标记物（活性GPIIb/IIIa和p -选择素）评估灌注血中下游血小板激活情况。乙酰水杨酸不能减少剪切诱导的血小板对所有三种结合蛋白的粘附。GPIIb/IIIa抑制剂（替罗非班和依替巴肽）显著降低血小板对纤维蛋白原的粘附。抗体阻断vWf或GPIbα可有效降低血小板对所有三种捕获蛋白的粘附以及灌注血中血小板的活化，表明vWf - GPIbα相互作用在介导剪切诱导的血小板聚集中起重要作用。
We have developed a microfluidic system to perfuse whole blood through a flow channel with an upstream stenotic region and a downstream protein capture region. This flow-based system was used to assay how effectively antiplatelet agents suppress shear-induced platelet adhesion and activation downstream of the stenotic region. Microcontact printing was used to covalently attach one of three platelet binding proteins [fibrinogen, collagen, or von Willebrand factor (vWf)] to the surface of the downstream capture region. Whole blood with an antiplatelet agent was transiently exposed to an upstream high wall shear rate (either 4860 s−1 or 11 560 s−1), and subsequently flowed over the downstream capture region where the platelet adhesion was measured. Several antiplatelet agents (acetylsalicylic acid, tirofiban, eptifibatide, anti-vWf, and anti-GPIbα) were evaluated for their efficacy in attenuating downstream adhesion. Following antibody blocking of vWf or GPIbα, downstream platelet activation was also assessed in perfused blood by flow cytometry using two activation markers (active GPIIb/IIIa and P-selectin). Acetylsalicylic acid demonstrated its inability to diminish shear-induced platelet adhesion to all three binding proteins. GPIIb/IIIa inhibitors (tirofiban and eptifibatide) significantly reduced platelet adhesion to fibrinogen. Antibody blocking of vWf or GPIbα effectively diminished platelet adhesion to all three capture proteins as well as platelet activation in perfused blood, indicating an essential role of vWf–GPIbα interaction in mediating shear-induced platelet aggregation.