Institute of Inflammation and Ageing, University of Birmingham , Birmingham , UK.
Institute of Cardiovascular Sciences, Institute for Biomedical Research, University of Birmingham , Birmingham , UK.
Platelets. 2019;30(7):893-900. doi: 10.1080/09537104.2018.1535704. Epub 2018 Oct 26.
The Total Thrombus-formation Analyser System (T-TAS) is a whole blood flow chamber system for the measurement of thrombus formation under variable shear stress conditions. Our current study sought to evaluate the potential utility of the T-TAS for the measurement of thrombus formation within human and mouse whole blood. T-TAS microchips (collagen, PL chip; collagen/tissue thromboplastin, AR chip) were used to analyze platelet (PL) or fibrin-rich thrombus formation, respectively. Blood samples from humans (healthy and patients with mild bleeding disorders) and wild-type (WT), mice were tested. Light transmission lumi-aggregometer (lumi-LTA) was performed in PRP using several concentrations of ADP, adrenaline, arachidonic acid, collagen, PAR-1 peptide and ristocetin. Thrombus growth (N = 22) increased with shear within PL (4:40 ± 1.11, 3:25 ± 0.43 and 3:12 ± 0.48 mins [1000, 1500 and 2000s]) and AR chips (3:55 ± 0.42 and 1:49 ± 0.19 [240s and 600s]). The area under the curve (AUC) on the PL chip was also reduced at 1000s compared to 1500/2000s (260 ± 51.7, 317 ± 55.4 and 301 ± 66.2, respectively). In contrast, no differences in the AUC between 240s and 600s were observed in the AR chip (1593 ± 122 and 1591 ± 158). The intra-assay coefficient of variation (CV) (n = 10) in the PL chip (1000s) and AR chip (240s) were T14.1%, T16.7%, T22.8% and AUC24.4% or T 9.03%, T8.64%, T23.8% and AUC5.1%. AR chip thrombus formation was inhibited by rivaroxaban (1 µM), but not with ticagrelor (10 µM). In contrast, PL chip thrombus formation was totally inhibited by ticagrelor. T-TAS shows an overall agreement with lumi-LTA in 87% of patients (n = 30) with normal PL counts recruited into the genotyping and phenotyping of platelet (GAPP) study and suspected to have a PL function defect. The onset (T) of thrombus formation in WT mice (N = 4) was shorter when compared to humans e.g. PL chip (1000s) T were 02:02 ± 00:23 and 03:30 ± 0:45, respectively). T-TAS measures thrombus formation and can be used for monitoring antithrombotic therapy, investigating patients with suspected PL function defects and monitoring PL function within mice.
全血栓形成分析系统（T-TAS）是一种用于在可变切应力条件下测量血栓形成的整体血液流动室系统。我们目前的研究旨在评估 T-TAS 在测量人类和小鼠全血中血栓形成的潜在用途。使用 T-TAS 微芯片（胶原，PL 芯片；胶原/组织凝血活酶，AR 芯片）分别分析血小板（PL）或富含纤维蛋白的血栓形成。测试了来自健康人和轻度出血障碍患者以及野生型（WT）小鼠的血液样本。使用几种浓度的 ADP、肾上腺素、花生四烯酸、胶原、PAR-1 肽和瑞斯托菌素在 PRP 中进行光传输 lumi-aggregometer（lumi-LTA）。在 PL 芯片中，血栓生长（N=22）随剪切力增加而增加（4:40±1.11、3:25±0.43 和 3:12±0.48 分钟[1000、1500 和 2000s]）和 AR 芯片（3:55±0.42 和 1:49±0.19[240s 和 600s]）。与 1500/2000s 相比，PL 芯片上的曲线下面积（AUC）在 1000s 时也减少（分别为 260±51.7、317±55.4 和 301±66.2）。相比之下，AR 芯片中 240s 和 600s 之间的 AUC 没有差异（1593±122 和 1591±158）。PL 芯片（1000s）和 AR 芯片（240s）的内测定变异系数（CV）（n=10）分别为 T14.1%、T16.7%、T22.8%和 AUC24.4%或 T9.03%、T8.64%、T23.8%和 AUC5.1%。AR 芯片中的血栓形成被 rivaroxaban（1µM）抑制，但 ticagrelor（10µM）则没有。相比之下，PL 芯片中的血栓形成完全被 ticagrelor 抑制。T-TAS 在被招募到血小板基因分型和表型（GAPP）研究并怀疑存在 PL 功能缺陷的 87%（n=30）正常 PL 计数的患者中与 lumi-LTA 总体一致。WT 小鼠（N=4）的血栓形成起始（T）时间比人类短，例如 PL 芯片（1000s）T 分别为 02:02±00:23 和 03:30±0:45）。T-TAS 可测量血栓形成，可用于监测抗血栓治疗、研究疑似 PL 功能缺陷患者以及监测小鼠中的 PL 功能。
