Cardiac-targeting transfection of tissue-type plasminogen activator gene to prevent the graft thrombosis and vascular anastomotic restenosis after coronary bypass.

Aim: To observe the tissue-type plasminogen activator gene (t-PA) plasmid packaged with albumin nanoparticles crosslinked to albumin ultrasound microbubbles for targeting transfection to myocardium to prevent the graft thrombosis and vascular anastomotic restenosis after coronary bypass. Methods: A dog model of coronary bypass using the autoallergic saphenous vein as the graft was made. A highly expressive t-PA gene plasmid packaged with albumin nanoparticles crosslinked to albumin ultrasound microbubbles was constructed. Targeting myocardial transfection was performed with this gene vector under the aid of therapeutic ultrasound(1 MHz, 1.5 w/cm2, 6 minutes, intravenously) after the bypass. The expression of t-PA in myocardium was detected with a multiclonal antibody to t-PA by the indirect immunohistochemical method. Venous blood t-PA and D-dimer contents were tested before and 1, 2 and 4 weeks after the operation. The effects of this gene vector on thrombosis of the grafts and the coronary intimal hyperplasia around the anastomotic stoma were observed using a routine pathological examination, a morphometry for intimal thickness and area and the immuno-histochemical stain with a monoclonal antibody to PCNA for estimating the intimal SMC proliferation. Results: The effective expression of t-PA protein by myocardium was obtained, followed by the persistent raises of blood t-PA and D-dimer 1, 2 and 4 weeks after the transfection. Thrombosis of the grafts was successfully restrained. The expression of PCNA by coronary intimal vSMCs and intimal hyperplasia were remarkablely reduced. Conclusion: This t-PA gene targeting vector could be used to prevent the dog thrombosis, which provided the experimental identification for prevention on human thrombotic diseases. (C) 2014 Elsevier Ltd. All rights reserved.