Experimental study of the preparation of targeted microbubble contrast agents carrying urokinase and RGDS.

PURPOSE

To explore the feasibility and the best combined proportion of the preparation of targeted and thrombolytic contrast agents carrying urokinase (UK) and Arg-Gly-Asp-Ser (RGDS), and to study its effect of thrombolysis in vitro.

METHODS

Urokinase and RGDS were combined to the surface of SonoVue by direct conjugation method. According to the different ratio of urokinase and RGDS when mixed, they were divided into three groups whose urokinase/RGDS were 1:1, 2:1 and 1:2, respectively. To measure the binding rate of microbubbles and urokinase as well as RGDS by flow cytometry. To detect the effect of thrombolysis by thrombolytic experiment in vitro. To detect the targeting effects by experiment in vivo.

RESULTS

The results of flow cytometry detection showed that the binding rates of urokinase and RGDS of three groups were, respectively, 73.4+/-11.0% and 67.1+/-10.9%, 8.8+/-7.2% and 7.8+/-6.9%, 49.7+/-21.3% and 45.9+/-21.7% after standing for 2h. In vitro thrombolysis experiment indicated that the urokinase had activity in the prepared contrast agent which the binding rates of urokinase and RGDS were the highest. And it was injected intravenous, the contrast agent aggregated on the surface of the thrombus of the rabbit femoral arterial. The thrombus emitted fluorescence.

CONCLUSIONS

The binding rate of the targeted contrast agent prepared by 1:1 of urokinase/RGDS was the highest. It had thrombolysis ability in vitro, and it had thrombo-targeting effect in vivo.