Procedure for quantification of platelet adhesion to biomaterials by radioscintigraphy

Abstract

Detection of adhered platelets on biomaterial surface that has blood-contacting application is an important test to assess its thrombogenicity. Usually, for measurement of platelet adhesion, after exposure to platelet-rich plasma (PRP) under standardized conditions the test surface is rinsed to remove non-adherent cells and is analyzed under scanning electron microscopy (SEM) to detect morphology of adhered cell and degree of aggregate formation. However, being a qualitative test it is unlikely to give an accurate estimate of platelets adhered to the surface. On the other hand, use of radiolabels enables quantification of platelets deposited on a material or device. Because of high gamma emission of 111In, it can be used for radioscintigraphy, however, its short half life (2.5 days) is a major hurdle in using it for quantification of platelet adhesion. 125I is a relatively strong radiolabel that is easily tagged to most of the proteins and has a relatively long half-life (60 days). The major objectives of this study are to standardize the labeling conditions to get good 125I activity on platelets, while maintaining normal cell function after they are labeled. Considering all possible uncertainties, quantity of isotope and platelets to be used and the conditions of iodination reaction are established to get repeatable and reproducible labeling of platelets. Further, it is demonstrated that 125I-platelets can be used to determine total number of cells adhered to titanium surface, which is known to be used as a blood-contacting biomaterial.