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用于诱导和分析动态剪切介导的血小板活化的基于微流体流动的平台——与标准化血液动力学剪切装置的初步验证

Microfluidic flow-based platforms for induction and analysis of dynamic shear-mediated platelet activation-Initial validation versus the standardized hemodynamic shearing device.

作者信息

Dimasi Annalisa, Roka-Moiia Yana, Consolo Filippo, Rasponi Marco, Fiore Gianfranco B, Slepian Marvin J, Redaelli Alberto

机构信息

Department of Electronics, Information and Bioengineering, Politecnico di Milano, Via Golgi 39, 20133 Milano, Italy.

Department of Medicine and Biomedical Engineering, Sarver Heart Center, University of Arizona, 1501 N Campbell Ave, Tucson, Arizona 85724, USA.

出版信息

Biomicrofluidics. 2018 May 22;12(4):042208. doi: 10.1063/1.5024500. eCollection 2018 Jul.

Abstract

A microfluidic flow-based platform (μFP), able to stimulate platelets via exposure of shear stress patterns pertinent to cardiovascular devices and prostheses, was compared to the Hemodynamic Shearing Device (HSD)-a state-of-the-art bench-top system for exposure of platelets to defined levels and patterns of shear. Platelets were exposed to time-varying shear stress patterns in the two systems; in detail, platelets were recirculated in the μFP or stimulated in the HSD to replicate comparable exposure time. Shear-mediated platelet activation was evaluated via (i) the platelet activity state assay, allowing the measurement of platelet-mediated thrombin generation and associated prothrombotic tendencies, (ii) scanning electron microscopy to evaluate morphological changes of sheared platelets, and (iii) flow cytometry for the determination of platelet phosphatidylserine exposure as a marker of shear activation. The results revealed good matching and comparability between the two systems, with similar trends of platelet activation, formation of microaggregates, and analogous trends of activation marker exposure for both the HSD and microfluidic-stimulated samples. These findings support future translation of the microfluidic platform as a Point-of-Care facsimile system for the diagnosis of thrombotic risk in patients implanted with cardiovascular devices.

摘要

一种基于微流体流动的平台(μFP),能够通过施加与心血管装置和假体相关的剪切应力模式来刺激血小板,该平台与血流动力学剪切装置(HSD)进行了比较,HSD是一种用于使血小板暴露于特定水平和模式剪切力的先进台式系统。在这两种系统中,血小板都暴露于随时间变化的剪切应力模式下;具体而言,血小板在μFP中循环或在HSD中受到刺激,以复制相当的暴露时间。通过以下方式评估剪切介导的血小板活化:(i)血小板活性状态测定,用于测量血小板介导的凝血酶生成及相关的血栓形成倾向;(ii)扫描电子显微镜,以评估剪切后血小板的形态变化;(iii)流式细胞术,用于测定血小板磷脂酰丝氨酸暴露情况,作为剪切活化的标志物。结果显示,这两种系统之间具有良好的匹配性和可比性,HSD和微流体刺激样本的血小板活化趋势、微聚集体形成以及活化标志物暴露趋势相似。这些发现支持了微流体平台作为一种即时护理模拟系统在植入心血管装置患者血栓形成风险诊断方面的未来转化应用。

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