Sabino Roberta M, Popat Ketul C
School of Advanced Materials Discovery, Colorado State University, Fort Collins, USA.
School of Biomedical Engineering, Colorado State University, Fort Collins, USA.
Bio Protoc. 2020 Feb 5;10(3):e3505. doi: 10.21769/BioProtoc.3505.
Biomaterial-associated thrombosis is still a major concern for blood-contacting implants. After the medical device is implanted and comes in contact with blood, several complex reactions occur, which may lead to thrombus formation and failure of the device. Therefore, it is essential to evaluate the biomaterial interaction with the whole blood. Several studies have been reported in the literature that evaluate different steps in the coagulation cascade, such as protein adsorption, plasma activation, and platelet adhesion , however, evaluation of whole blood clotting on biomaterial surfaces is not widely reported. Here, a protocol to evaluate whole blood clotting on 2D biomaterials surfaces via a simple and fast hemolysis assay is presented. Whole human blood is placed onto the biomaterial surfaces and is allowed to clot for different time periods. After the specific time intervals, the surfaces are transferred into deionized (DI) water to release the free hemoglobin and the absorbance of this solution is measured. The absorbance value is proportional to the free hemoglobin concentration in the DI water due to lysis of red blood cells and gives an indirect correlation to the extent of blood clotting on the biomaterial surfaces. This protocol provides a fast, facile and effective method to measure the anti-thrombogenic properties of biomaterials.
生物材料相关的血栓形成仍然是血液接触植入物的一个主要问题。在医疗设备植入并与血液接触后，会发生几种复杂的反应，这可能导致血栓形成和设备失效。因此，评估生物材料与全血的相互作用至关重要。文献中已经报道了几项评估凝血级联反应中不同步骤的研究，如蛋白质吸附、血浆激活和血小板粘附，然而，关于生物材料表面全血凝血的评估报道并不广泛。在此，我们提出了一种通过简单快速的溶血试验来评估二维生物材料表面全血凝血的方案。将全人类血液放置在生物材料表面，并使其在不同时间段内凝血。在特定时间间隔后，将表面转移到去离子（DI）水中以释放游离血红蛋白，并测量该溶液的吸光度。由于红细胞的裂解，吸光度值与去离子水中游离血红蛋白的浓度成正比，并与生物材料表面的血液凝固程度间接相关。该方案提供了一种快速、简便且有效的方法来测量生物材料的抗血栓形成特性。
