Department of Laboratory Medicine, Keio University School of Medicine, Tokyo, Japan.
Thromb Res. 2013 Aug;132(2):263-70. doi: 10.1016/j.thromres.2013.05.026. Epub 2013 Jun 16.
A whole blood flow-chamber system, the Total Thrombus-formation Analysis System (T-TAS), was developed for quantitative analysis of platelet thrombus formation (PTF) using microchips with thrombogenic surfaces (collagen, PL chip; collagen plus tissue thromboplastin, AR chip) under shear stress conditions. We evaluated the usefulness of the T-TAS for assessing individual thrombogenicity compared with other platelet function tests.
Blood samples from 31 healthy volunteers were applied to the T-TAS to measure PTF starting time (T10: time to reach 10 kPa), occlusion time (T60 for PL chip; T80 for AR chip), and area under the curve (AUC10, area under curve until 10 min for PL chip; AUC30, 30 min for AR chip) under various shear rates (1000, 1500, 2000s(-1) for PL chip; 300 s(-1) for AR chip). Platelet functions were also tested using platelet aggregometry, the PFA-100 (collagen and epinephrine [C/EPI], collagen and adenosine diphosphate [C/ADP]), and the VerifyNow P2Y12 assay.
Individual pressure waveforms, including PTF starting and ending points, varied among healthy subjects. In the PL chip, T10 and AUC10 showed a shear-dependent correlation with C/EPI or C/ADP. VerifyNow P2Y12 values were not significantly associated with the parameters of the T-TAS. Platelet counts were correlated with all AR measurements, and mostly with PL measurements.
The results of the T-TAS were associated with those of the PFA-100 in many respects, indicating that its characteristics are related to shear-induced PTF. The T-TAS showed few correlations with platelet aggregometry and the VerifyNow P2Y12 assay. The T-TAS may allow for the measurement of comprehensive parameters of individual thrombogenicity under whole blood flow conditions.
我们开发了一种全血流动室系统,即全血栓形成分析系统(T-TAS),用于在切应力条件下使用具有血栓形成表面(胶原、PL 芯片;胶原加组织凝血活酶、AR 芯片)的微芯片定量分析血小板血栓形成(PTF)。我们评估了 T-TAS 用于评估个体血栓形成倾向的有用性,与其他血小板功能测试进行了比较。
将 31 名健康志愿者的血液样本应用于 T-TAS,以在各种切变率(PL 芯片的 1000、1500、2000s(-1);AR 芯片的 300 s(-1))下测量 PTF 起始时间(T10:达到 10 kPa 的时间)、闭塞时间(PL 芯片的 T60;AR 芯片的 T80)和曲线下面积(PL 芯片的 AUC10,直到 10 分钟的曲线下面积;AR 芯片的 AUC30,30 分钟)。还使用血小板聚集仪、PFA-100(胶原和肾上腺素[C/EPI],胶原和二磷酸腺苷[C/ADP])和 VerifyNow P2Y12 测定法测试血小板功能。
个体压力波形,包括 PTF 起始和结束点,在健康受试者中存在差异。在 PL 芯片中,T10 和 AUC10 与 C/EPI 或 C/ADP 呈切变依赖性相关。VerifyNow P2Y12 值与 T-TAS 的参数没有显著相关性。血小板计数与所有 AR 测量值相关,与 PL 测量值大多相关。
T-TAS 的结果在许多方面与 PFA-100 的结果相关,表明其特征与剪切诱导的 PTF 有关。T-TAS 与血小板聚集仪和 VerifyNow P2Y12 测定法的相关性较小。T-TAS 可能允许在全血流动条件下测量个体血栓形成倾向的综合参数。