A simple assay to detect endothelial cell injury; measurement of released thrombomodulin from cells

Abstract

A simple assay to determine the degree of endothelial cell injury has been developed using released thrombomodulin as the index. Thrombomodulin is a cell surface protein on endothelial cells which is released from the cell upon injury. In this assay, bovine arterial endothelial cells were cultured in serum free medium with the test substances and the amount of thrombomodulin released into the culture medium was measured by enzyme immunoassay. Substances which are known to injure cells such as H2O2, prostaglandin A2, lipopolysaccharide, and elastase released significant amounts of thrombomodulin. The sensitivity of this assay for mild injury was superior or at least equal to the traditional 51Cr release method. Since this method does not require the use of radioisotopes, it seems to be advantageous and suitable for the detection of endothelial cell injury during routine examination.