Efficacy of plasmin, microplasmin, and streptokinase-plasmin complex for the in vitro degradation of fibronectin and laminin- implications for vitreoretinal surgery

Abstract

Purpose: Plasmin enzyme generates vitreoretinal separation by degradation of laminin and fibronectin in the vitreoretinal interface. It can be activated from plasminogen by urokinase, tissue plasminogen activator, or by formation of a 1:1 complex with streptokinase. The latter is then converted into a streptokinase-plasmin-complex (SK-P), which displays fibrinolytic activity and can generate free plasmin by proteolysis of plasminogen. We compared the efficacy of SK-P, SK-P activated plasmin, urokinase activated plasmin (UK-P), and microplasmin, a truncated form of plasmin, in cleaving laminin and fibronectin.

Methods: Streptokinase (SK) was added to human plasminogen in molar ratios between 1:100 and 2:1, generating SK-P at ratios > 1:1, and mixtures of SK-P and free plasmin (SK-P/plasmin) at lower ratios. SK-PL, SK-P/plasmin, UK-P, and microplasmin were added to laminin and fibronectin, incubated at 37 degrees C for 30 min-22 hr and processed for SDS-PAGE.

Results: Proteolysis using SK-activated plasminogen increased when the SK/plasminogen ratio was decreased, generating increasing amounts of free plasmin. Microplasmin and urokinase-activated plasmin displayed similar proteolysis of both laminin and fibronectin as SK/plasminogen at ratios of 1:10 or lower.

Conclusion: The mode of plasminogen activation influences the efficacy of proteolysis for laminin and fibronectin and should be considered when plasmin is used in vitreoretinal surgery.