Department of Physiology, College of Medicine, National Cheng-Kung University, Tainan, 70101, ROC, Taiwan, Fax: 886-6-2362780.
Platelets. 1992;3(3):145-53. doi: 10.3109/09537109209013175.
A flow system consisting of a parallel-plate flow chamber mounted on the epifluorescence video microscope has been constructed to allow direct visualization of the entire platelet adhesion process under whole blood flow conditions. Adhered platelets with recorded adhesion history were individually identified and observed in detail using a scanning electron microscope. In this study we used cover glasses coated with fibrinogen, fibrin, or collagen as the testing surface. From experiments carried out at the surface shear rate of 445 s(-1), we found that (1) platelet adhesion was a dynamic process that involved attaching, detaching, relocation and transient contact; (2) platelets adhered to all three types of protein-coated surfaces with platelet adhesion on collagen being most unstable; (3) most of these adhered platelets immediately formed short pseudopods after surface contact; (4) platelets adhered to fibrinogen or fibrin were basically non-overlapping and they underwent further shape change with increasing number /length of pseudopods and increasing extent of cytoplasmic spreading; (5) on collagen-coated surface most incoming platelets attached to previously adhered platelets rather than to the collagen threads for blood-surface contact times longer than 30 s; (6) these platelets formed multicellular thrombi with largest thrombi located at about 0.2-0.4 mm from the upstream edge and (7) platelets in the thrombi formed numerous short pseudopods and started fusing with one another within 2 min. These observations show that platelet adhesion under blood flow is a complex and dynamic process and that adhered platelets undergo heterogeneous post-contact morphological changes. Moreover, our results indicate that fibrinogen and fibrin coatings are adhesive while collagen coating is most stimulatory to platelets.
已构建了一个由安装在落射荧光显微镜上的平行板流动室组成的流动系统,以允许在全血流条件下直接观察整个血小板黏附过程。使用扫描电子显微镜对具有记录的黏附历史的黏附血小板进行单独识别和详细观察。在这项研究中,我们使用涂覆有纤维蛋白原、纤维蛋白或胶原蛋白的盖玻片作为测试表面。在表面剪切速率为 445 s(-1) 的实验中,我们发现:(1)血小板黏附是一个动态过程,涉及附着、脱离、重定位和瞬时接触;(2)血小板黏附于所有三种蛋白质涂覆表面,其中黏附于胶原蛋白的血小板最不稳定;(3)这些黏附的血小板在与表面接触后立即形成短伪足;(4)黏附于纤维蛋白原或纤维蛋白的血小板基本上不重叠,并且随着伪足数量/长度的增加和细胞质扩散程度的增加而进一步发生形状变化;(5)在胶原蛋白涂覆的表面上,对于血液-表面接触时间长于 30 s 的情况,大多数进入的血小板附着于先前黏附的血小板,而不是附着于胶原蛋白丝;(6)这些血小板形成多细胞血栓,最大的血栓位于距上游边缘约 0.2-0.4 mm 处;(7)血栓中的血小板形成许多短伪足,并在 2 分钟内开始彼此融合。这些观察结果表明,血流中的血小板黏附是一个复杂而动态的过程,并且黏附的血小板在接触后会发生异质的形态变化。此外,我们的结果表明,纤维蛋白原和纤维蛋白涂层具有黏附性,而胶原蛋白涂层对血小板最具刺激性。