Immobilization of plasmid DNA on an anti-DNA antibody modified coronary stent for intravascular site-specific gene therapy

摘要

Background The aim of the present study was to investigate the incorporation of plasmid DNA (pDNA) onto a coronary stent by chemo-immunoconjugation for achieving site-specific gene delivery. Methods Anti-DNA immunoglobulin M antibody was chemically linked onto collagen-coated stent by using N-succinimidyl-3-(2-pyridyldithiol)-propionate as cross-linker. pDNA was tethered on the antibody-immobilized stent by highly specific antigen-antibody affinity interaction. Radioactive-labeled antibody and pDNA were used to evaluate binding capacity and stability. A reporter plasmid pEGFP was tethered on the antibody-immobilized stents that was assessed in cell culture and in rabbit carotid model. Results The amount of antibody chemically linked on the stents was 15-fold higher than that of the control and its retention time was also significantly longer. The pEGFP-tethered stents had no detrimental effects on cell growth. In cell culture studies, numerous green fluorescent protein (GFP)-transfected cells were only found on the stent, which demonstrated high localization and efficiency of gene delivery. The overall GFP transfection efficiency in treated rabbit carotid arteries was 2.8 +/- 0.7% of the total cells. However, the rate of neointima transfection was 7.0 +/- 0.8% of total cells in this region. importantly, no distal spreading of the vector was detected by polymerase chain reaction, either in distal organs or in the downstream segments of the stented arteries. Conclusions For the first time, our group reports the successful use of anti-DNA antibody-immobilized metal stent as plasmid gene delivery system that possess high efficiency and site- specificity in vitro and in vivo. Copyright (c) 2008 John Wiley & Sons, Ltd.