Bisphosphonate-mediated gene vector delivery from the metal surfaces of stents.
The clinical use of metallic expandable intravascular stents has resulted in improved therapeutic outcomes for coronary artery disease. However, arterial reobstruction after stenting, in-stent restenosis, remains an important problem. Gene therapy to treat in-stent restenosis by using gene vector delivery from the metallic stent surfaces has never been demonstrated. The present studies investigated the hypothesis that metal-bisphosphonate binding can enable site-specific gene vector delivery from metal surfaces. Polyallylamine bisphosphonate (PAA-BP) was synthesized by using Michael addition methodology. Exposure to aqueous solutions of PAA-BP resulted in the formation of a monomolecular bisphosphonate layer on metal alloy surfaces (steel, nitinol, and cobalt-chromium), as demonstrated by x-ray photoelectron spectroscopy. Surface-bound PAA-BP enabled adenoviral (Ad) tethering due to covalent thiol-binding of either anti-Ad antibody or a recombinant Ad-receptor protein, D1. In arterial smooth muscle cell cultures, alloy samples configured with surface-tethered Ad were demonstrated to achieve site-specific transduction with a reporter gene, (GFP). Rat carotid stent angioplasties using metal stents exposed to aqueous PAA-BP and derivatized with anti-knob antibody or D1 resulted in extensive localized Ad-GFP expression in the arterial wall. In a separate study with a model therapeutic vector, Ad-inducible nitric oxide synthase (iNOS) attached to the bisphosphonate-treated metal stent surface via D1, significant inhibition of restenosis was demonstrated (neointimal/media ratio 1.68 +/- 0.27 and 3.4 +/- 0.35; Ad-iNOS vs. control, P < 0.01). It is concluded that effective gene vector delivery from metallic stent surfaces can be achieved by using this approach.
Fishbein
Ilia
I
Division of Cardiology, The Children's Hospital of Philadelphia, Department of Chemical, Philadelphia, PA 19104, USA.
Alferiev
Ivan S
IS
Nyanguile
Origene
O
Gaster
Richard
R
Vohs
John M
JM
Wong
Gordon S
GS
Felderman
Howard
H
Chen
I-Wei
IW
Choi
Hoon
H
Wilensky
Robert L
RL
Levy
Robert J
RJ
D000256:Adenoviridae / Q000378:metabolism; D017130:Angioplasty / Q000379:methods*; D000818:Animals; D002478:Cells, Cultured; D003324:Coronary Artery Disease / Q000150:complications / Q000601:surgery*; D004164:Diphosphonates / Q000378:metabolism*; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000378:metabolism / Q000627:therapeutic use*; D006083:Graft Occlusion, Vascular / Q000209:etiology / Q000628:therapy*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008297:Male; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D011073:Polyamines / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013057:Spectrum Analysis; D015607:Stents; D014161:Transduction, Genetic / Q000379:methods
D004164:Diphosphonates; D011073:Polyamines; D049452:Green Fluorescent Proteins; C063994:polyallylamine; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat