ULTRASOUND ENHANCEMENT OF THROMBOLYSIS AND REPERFUSION INVITRO

Objectives. The aims of this study were 1) to develop an in vitro flow system in which reperfusion mediated by ultrasound-accelerated thrombolysis could be studied, and 2) to test whether ultrasound-accelerated thrombolysis could hasten reperfusion in this system. Background. Ultrasound has been shown to increase tissue plasminogen activator (t-PA)-induced thrombolysis in vitro as assessed by radioactive fibrinogen release from labeled clots and in an animal in vivo model. Methods. To test whether reperfusion is accelerated, we created obstructive whole blood clots in an in vitro flow system. Four control clots were exposed to ultrasound only without any thrombolytic agent (group 1). Sixteen clots were exposed to continuous infusion of recombinant tissue-type plasminogen activator rt-PA and randomized to either continuous wave ultrasound exposure at a frequency of 0.5 MHz and an intensity of 8 W/cm2 (group 2) or to no ultrasound (group 3). Flow distal to the clot and the rate of release of radiolabeled fibrin products were used as an index of reperfusion and thrombolysis, respectively. Samples were obtained for measurements of lytic variables such as plasminogen, fibrinogen and rt-PA concentrations. Results. Flow was significantly higher in the rt-PA-treated clots within 10 min of exposure to ultrasound than in those without such exposure (9.4 +/- 9.9% of maximal flow in group 2 vs. 0.5 +/- 1.5% in group 3, p < 0.05). The maximal difference in flow between groups 2 and 3 was achieved at 25 min (61.0 +/- 30.4% vs. 14.2 +/- 14.7%, p = 0.03). Thrombolysis was significantly higher after 15 min of ultrasound exposure (12.8 +/- 9.1% in the ultrasound-treated group 2 vs. 4.0 +/- 3.9% in group 3, p < 0.05). The maximal difference between groups 2 and 3 occurred at 25 min (26.7 +/- 13.1% vs. 7.24 +/- 5.7%, p < 0.004). Neither flow nor clot lysis occurred in group 1. Plasminogen and fibrinogen concentrations and rt-PA antigen concentrations were consistent with those observed during fibrinolytic therapy in vivo. Conclusions. Continuous wave ultrasound at 0.5 MHz and an intensity of 8 W/cm2 accelerates rt-PA-induced thrombolysis and reperfusion in vitro.