Potter Timothy M, Rodriguez Jamie C, Neun Barry W, Ilinskaya Anna N, Cedrone Edward, Dobrovolskaia Marina A
Cancer Research Technology Program¸ Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA.
Methods Mol Biol. 2018;1682:103-124. doi: 10.1007/978-1-4939-7352-1_10.
Blood clotting is a complex process which involves both cellular and biochemical components. The key cellular players in the blood clotting process are thrombocytes or platelets. Other cells, including leukocytes and endothelial cells, contribute to clotting by expressing the so-called pro-coagulant activity (PCA) complex on their surface. The biochemical component of blood clotting is represented by the plasma coagulation cascade, which includes plasma proteins also known as coagulation factors. The coordinated interaction between platelets, leukocytes, endothelial cells, and plasma coagulation factors is necessary for maintaining hemostasis and for preventing excessive bleeding. Undesirable activation of all or some of these components may lead to pathological blood coagulation and life-threatening conditions such as consumptive coagulopathy or disseminated intravascular coagulation (DIC). In contrast, unintended inhibition of the coagulation pathways may lead to hemorrhage. Thrombogenicity is the property of a test material to induce blood coagulation by affecting one or more elements of the clotting process. Anticoagulant activity refers to the property of a test material to inhibit coagulation. The tendency to cause platelet aggregation, perturb plasma coagulation, and induce leukocyte PCA can serve as an in vitro measure of a nanomaterial's likelihood to be pro- or anticoagulant in vivo. This chapter describes three procedures for in vitro analyses of platelet aggregation, plasma coagulation time, and activation of leukocyte PCA. Platelet aggregation and plasma coagulation procedures have been described earlier. The revision here includes updated details about nanoparticle sample preparation, selection of nanoparticle concentration for the in vitro study, and updated details about assay controls. The chapter is expanded to describe a method for the leukocyte PCA analysis and case studies demonstrating the performance of these in vitro assays.
血液凝固是一个复杂的过程，涉及细胞和生化成分。血液凝固过程中的关键细胞成分是血小板。其他细胞，包括白细胞和内皮细胞，通过在其表面表达所谓的促凝血活性（PCA）复合物来促进凝血。血液凝固的生化成分由血浆凝血级联反应代表，其中包括也被称为凝血因子的血浆蛋白。血小板、白细胞、内皮细胞和血浆凝血因子之间的协同相互作用对于维持止血和防止过度出血是必要的。这些成分中全部或部分的不良激活可能导致病理性血液凝固和危及生命的状况，如消耗性凝血病或弥散性血管内凝血（DIC）。相反，凝血途径的意外抑制可能导致出血。血栓形成性是指测试材料通过影响凝血过程的一个或多个要素来诱导血液凝固的特性。抗凝血活性是指测试材料抑制凝血的特性。引起血小板聚集、干扰血浆凝血和诱导白细胞PCA的倾向可作为纳米材料在体内具有促凝血或抗凝血可能性的体外测量指标。本章描述了三种用于体外分析血小板聚集、血浆凝血时间和白细胞PCA激活的方法。血小板聚集和血浆凝血方法先前已作描述。此处的修订包括关于纳米颗粒样品制备的更新细节、体外研究中纳米颗粒浓度的选择以及测定对照的更新细节。本章进行了扩展，以描述白细胞PCA分析方法以及证明这些体外测定性能的案例研究。
