Efficient adventitial gene delivery to rabbit carotid artery with cationic polymer-plasmid complexes.
Different lipids and cationic polymers were tested in vitro for their ability to transfect rabbit aortic smooth muscle cells and human endothelial cells with lacZ marker gene. Toxicity of the complexes was evaluated with MTT assay. Selected plasmid-polymer complexes with different charge ratios were then tested for in vivo gene transfer efficiency using adventitial gene transfer by placing a silastic gene delivery reservoir (collar) around the carotid artery. Transfection efficiency was determined by X-gal staining 3 days after the gene transfer. Based on in vitro experiments, fractured polyamidoamine dendrimers and polyethylenimines (PEI) were selected for in vivo experiments. Fractured dendrimers (generation 6, +/- charge ratio of 3) had the highest in vivo gene transfer efficiency (4.4% +/- 1.7). PEI with molecular size of 25 kDa (+/- charge ratio 4) was also effective (2.8% +/- 1.8) in this model. PEI of 800 kDa showed a constant but modest gene transfer efficiency (1.8% +/- 0.1) with all charge ratios. A low level gene transfer was also detected with naked DNA (0.5% +/- 0.3). No signs of inflammation were seen in any of the study groups. We show here that in vitro cell culture experiments can be used to identify efficient in vivo gene transfer methods for arterial gene therapy, but the charge ratios for each complex must be optimized in vivo. It is concluded that fractured dendrimer and PEI are efficient gene delivery vehicles and can be used for arterial gene therapy via adventitial gene delivery route.
Turunen
M P
MP
AI Virtanen Institute, University of Kuopio, Finland.
Hiltunen
M O
MO
Ruponen
M
M
Virkamäki
L
L
Szoka
F C
FC
Urtti
A
A
Ylä-Herttuala
S
S
D000818:Animals; D002339:Carotid Arteries / Q000033:anatomy & histology; D002412:Cations; D002478:Cells, Cultured; D004730:Endothelium, Vascular; D005822:Genetic Vectors; D008297:Male; D009131:Muscle, Smooth, Vascular; D010957:Plasmids; D011108:Polymers; D011817:Rabbits; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics*
D002412:Cations; D011108:Polymers; D001616:beta-Galactosidase