Adsorption of coagulation proteins and adhesion and activation of platelets at the blood-solid interface. An experimental study of human whole blood
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H Nygren1, J H Elam, M Stenberg
1Department of Histology, University of Göteborg, Sweden.
A procedure is presented allowing detailed studies of the adsorption of coagulation factors from whole blood on to surface. Anticoagulant (citrate or hirudin) was added to fresh venous blood. The blood was incubated in hydrophilic or hydrophobic glass tubes without contact with air. The adsorption of fibrinogen, fibronectin and factor IX was measured with an enzyme immunoassay using specific antibodies directed against these proteins. Adsorption of enzymically active kallikrein was measured using a chromogenic peptide substrate. Adhesion and activation of platelets was measured by direct examination in a scanning electron microscope and by measurement of release of beta-thromboglobulin. The results show that the adsorption of plasma proteins at the blood-solid interface is dependent on the anticoagulant used, surface energy of the test surface and incubation time. In experiments using hirudin a specific inactivator of thrombin, as anticoagulant, we found dynamic changes of the adsorbed protein film which could not be studied using citrated blood.
A procedure is presented allowing detailed studies of the adsorption of coagulation factors from whole blood on to surface. Anticoagulant (citrate or hirudin) was added to fresh venous blood. The blood was incubated in hydrophilic or hydrophobic glass tubes without contact with air. The adsorption of fibrinogen, fibronectin and factor IX was measured with an enzyme immunoassay using specific antibodies directed against these proteins. Adsorption of enzymically active kallikrein was measured using a chromogenic peptide substrate. Adhesion and activation of platelets was measured by direct examination in a scanning electron microscope and by measurement of release of beta-thromboglobulin. The results show that the adsorption of plasma proteins at the blood-solid interface is dependent on the anticoagulant used, surface energy of the test surface and incubation time. In experiments using hirudin a specific inactivator of thrombin, as anticoagulant, we found dynamic changes of the adsorbed protein film which could not be studied using citrated blood.