Heparin was covalently coupled to collageno-elastic grafts (CET) derived from lamb carotid arteries, by using the crosslinking agent 1-ethyl-3 (3-dimethyl-aminopropyl) carbodiimide (EDC). The collagenous grafts were pretreated with various aminating agents in order to enhance the number of available binding sites on the collagen surface. By varying the EDC/heparin weight ratio, the pH of the immobilization media, and the pretreatment agent, a global search pattern maximized heparin loading at 3.90 +/- 0.36 USP heparin/cm2 collagenous graft surface when the EDC/heparin ratio was 2:1 at a pH of 1.5 with 1 M hydroxylamine sulfate as the pretreatment agent. Heparinized CETs were superior to nonheparinized CETs by exhibiting both enhanced antiplatelet activity in using an in vitro differential recirculation reactor with chromium-51 tagged platelets and enhanced patency when interposed in canine carotid arteries. Both antiplatelet activity and patency duration for heparinized CETs were independent of heparin loading.
SummaryPlasmin was immobilized on collageno-elastic tubes (CET) using carbodiimide as the cross-linking agent. The effects of plasmin-CET grafts and corresponding soluble plasmin on fibrinogen, thrombin-mediated fibrinogen activation, and platelet activity, were investigatedThere was a significant increase in fibrinogen deposition on plasmin-CETs over non-plasmin (i.e. control) CETs. Furthermore, exposure of fibrinogen to plasmin CETs enhances its deposition to control grafts situated downstream. Plasmin-bound CETs retained higher platelet deposition when preliminarily coated with fibrinogen. Finally, plasmin exerted a positive effect on thrombin-mediated fibrinogen activation at low plasmin concentrations. A mechanistic hypothesis aimed at interpreting this
finding is proposed.
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