Surface characterization and platelet adhesion studies on polyethylene surface with hirudin immobilization.

Hirudin, a protein composed of 65 or 66 amino acid, is a newly risen anticoagulant agent and has been considered as the most potent thrombin inhibitor. Hirudin can block the active site of thrombin by means of its carboxylic acid reaction with the active regime of thrombin, and becomes a tightly bound complex, and thus controls the formation of thrombus. Hirudin was covalently immobilized onto the water-soluble carbodiimide preactivated and chromic acid oxidized PE surface. Surface chemistry analysis indicated that a certain amount of carboxylic acid groups was generated on the polyethylene surface after oxidation with chromic acid solution. The amount of carboxylic acid functional group increased with the oxidation time. In addition, polyethylene surface was etched by chromic acid solution, and a rougher surface was created. The morphology of oxidized polyethylene surface was similar to each other among the samples with oxidation time from 1 to 8 min. ESCA results indicated the number of hirudin molecules immobilized was constant at the reaction time studied. In vitro platelet adhesion assay indicated the number of adhered platelets on the oxidized polyethylene surface increased significantly after oxidation. In contrast, surface with hirudin immobilization showed a reduction in adhered platelet density than the chromic acid oxidized counterpart due to the decrease of platelet-activating capability by the hirudin-thrombin complex and the differences in the adsorbed protein composition. Copyright 2001 Kluwer Academic Publishers