Dimasi Annalisa, Rasponi Marco, Consolo Filippo, Fiore Gianfranco B, Bluestein Danny, Slepian Marvin J, Redaelli Alberto
Department of Electronics, Information and Bioengineering, Politecnico di Milano, Via Golgi 39, 20133, Milano, Italy.
Department of Electronics, Information and Bioengineering, Politecnico di Milano, Via Golgi 39, 20133, Milano, Italy.
Med Eng Phys. 2017 Oct;48:31-38. doi: 10.1016/j.medengphy.2017.08.005. Epub 2017 Aug 30.
Thrombus formation is a major adverse event affecting patients implanted with ventricular assist devices (VADs). Despite anti-thrombotic drug administration, thrombotic events remain frequent within the first year post-implantation. Platelet activation (PA) is an essential process underling thrombotic adverse events in VAD systems. Indeed, abnormal shear forces, correlating with specific flow trajectories of VADs, are strong agonists mediating PA. To date, the ability to determine efficacy of anti-platelet (AP) agents under shear stress conditions is limited. Here, we present a novel microfluidic platform designed to replicate shear stress patterns of a clinical VAD, and use it to compare the efficacy of two AP agents in vitro. Gel-filtered platelets were incubated with i) acetylsalicylic acid (ASA) and ii) ticagrelor, at two different concentrations (ASA: 125 and 250 µM; ticagrelor: 250 and 500 nM) and were circulated in the VAD-emulating microfluidic platform using a peristaltic pump. GFP was collected after 4 and 52 repetitions of exposure to the VAD shear pattern and tested for shear-mediated PA. ASA significantly inhibited PA only at 2-fold higher concentration (250 µM) than therapeutic dose (125 µM). The effect of ticagrelor was not dependent on drug concentration, and did not show significant inhibition with respect to untreated control. This study demonstrates the potential use of microfluidic platforms as means of testing platelet responsiveness and AP drug efficacy under complex and realistic VAD-like shear stress conditions.
血栓形成是影响植入心室辅助装置(VAD)患者的主要不良事件。尽管使用了抗血栓药物,但血栓事件在植入后的第一年内仍然频繁发生。血小板活化(PA)是VAD系统中血栓性不良事件的一个重要潜在过程。事实上,与VAD特定血流轨迹相关的异常剪切力是介导PA的强效激动剂。迄今为止,在剪切应力条件下确定抗血小板(AP)药物疗效的能力有限。在此,我们展示了一种新型微流控平台,该平台旨在复制临床VAD的剪切应力模式,并使用它在体外比较两种AP药物的疗效。将凝胶过滤血小板与i)乙酰水杨酸(ASA)和ii)替格瑞洛在两种不同浓度下孵育(ASA:125和250µM;替格瑞洛:250和500 nM),并使用蠕动泵在模拟VAD的微流控平台中循环。在重复暴露于VAD剪切模式4次和52次后收集绿色荧光蛋白(GFP),并测试剪切介导的PA。ASA仅在浓度比治疗剂量(125µM)高2倍(250µM)时才显著抑制PA。替格瑞洛的作用不依赖于药物浓度,与未治疗的对照组相比未显示出显著抑制作用。这项研究证明了微流控平台作为在复杂且逼真的类似VAD剪切应力条件下测试血小板反应性和AP药物疗效手段的潜在用途。