Antithrombogenicity of lumbrokinase-immobilized polyurethane.
Lumbrokinase is a potent fibrinolytic enzyme purified from the earthworm, Lumbricus rubellus. We immobilized 18 IU/cm2 of lumbrokinase to polyurethane using maleic anhydride methylvinyl ether copolymer (MAMEC) as an enzyme carrier, and the proteolytic and fibrinolytic activities of immobilized lumbrokinase were assayed. Immobilized lumbrokinase retained about 34% of its activity, compared with soluble lumbrokinase activity. Immobilized lumbrokinase showed stability against thermal inactivation and degradation and within a various pH range. The optimal pH of immobilized lumbrokinase shifted 1.0 pH unit upward compared with soluble enzyme. Upon exposure to the human whole blood, less amount of 125I-fibrinogen was adsorbed to lumbrokinase-immobilized surface than to the polyurethane control surface. The lumbrokinase-immobilized surface showed less platelet adhesion than did the MAMEC-grafted surface. At the early stage of platelet adhesion, the number of adhered platelets increased on the lumbrokinase-immobilized surface with increasing time; yet, the platelet number drastically decreased on the lumbrokinase-immobilized surface after 80 min incubation. This suggests that lumbrokinase-immobilized polyurethane digested the adsorbed fibrinogen and inhibited platelet adhesion on the surface, probably by inhibiting fibrinogen adsorption to be highly antithrombogenic. Clinical applications of this material to artificial organs should be developed in the near future.
Ryu
G H
GH
Department of Biomedical Engineering, College of Medicine, Seoul National University, Korea.
Park
S
S
Kim
M
M
Han
D K
DK
Kim
Y H
YH
Min
B
B
D000327:Adsorption; D010450:Endopeptidases / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000494:pharmacology*; D005337:Fibrin / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D006801:Humans; D006868:Hydrolysis; D010973:Platelet Adhesiveness / Q000502:physiology; D011140:Polyurethanes; D011506:Proteins / Q000378:metabolism
D004800:Enzymes, Immobilized; D005343:Fibrinolytic Agents; D011140:Polyurethanes; D011506:Proteins; D005337:Fibrin; D010450:Endopeptidases; C083421:lumbrokinase