title: Anchoring of self-assembled plasmid DNA/anti-DNA antibody/cationic lipid micelles on bisphosphonate-modified stent for cardiovascular gene delivery.
abstract: OBJECTIVE
  To investigate the anchoring of plasmid DNA/anti-DNA antibody/cationic lipid tri-complex (DAC micelles) onto bisphosphonate-modified 316 L coronary stents for cardiovascular site-specific gene delivery.
  
  
  METHODS
  Stents were first modified with polyallylamine bisphosphonate (PAA-BP), thereby enabling the retention of a PAA-BP molecular monolayer that permits the anchoring (via vector-binding molecules) of DAC micelles. DAC micelles were then chemically linked onto the PAA-BP-modified stents by using N-succinimidyl-3-(2-pyridyldithiol)-propionate (SPDP) as a crosslinker. Rhodamine-labeled DNA was used to assess the anchoring of DAC micelles, and radioactive-labeled antibody was used to evaluate binding capacity and stability. DAC micelles (encoding green fluorescent protein) were tethered onto the PAA-BP-modified stents, which were assessed in cell culture. The presence of a PAA-BP molecular monolayer on the steel surface was confirmed by X-ray photoelectron spectroscopy and atomic force microscope analysis.
  
  
  RESULTS
  The anchoring of DAC micelles was generally uniform and devoid of large-scale patches of defects. Isotopic quantification confirmed that the amount of antibody chemically linked on the stents was 17-fold higher than that of the physical adsorbed control stents and its retention time was also significantly longer. In cell culture, numerous green fluorescent protein-positive cells were found on the PAA-BP modified stents, which demonstrated high localization and efficiency of gene delivery.
  
  
  CONCLUSIONS
  The DAC micelle-immobilized PAA-BP-modified stents were successful as a gene delivery system. Gene delivery using DAC micelle-tethered stent-based PAA-BP functionalization should be suitable for a wide array of single or multiple therapeutic gene strategies, and could be used on cardiovascular metallic implants for achieving efficient gene therapy.
authors.affiliation: Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, People's Republic of China.
authors.forename: Guilei
authors.initials: G
authors.lastname: Ma
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Ilia
authors.initials: I
authors.lastname: Fishbein
authors.forename: Mei
authors.initials: M
authors.lastname: Yu
authors.forename: Linhua
authors.initials: L
authors.lastname: Zhang
authors.forename: Ivan S
authors.initials: IS
authors.lastname: Alferiev
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Cunxian
authors.initials: C
authors.lastname: Song
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Levy
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000974:Antibodies, Antinuclear / Q000737:chemistry / Q000378:metabolism; D002317:Cardiovascular Agents / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004164:Diphosphonates / Q000737:chemistry; D016503:Drug Delivery Systems / Q000295:instrumentation / Q000379:methods*; D054855:Drug-Eluting Stents; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008422:Materials Testing; D008823:Micelles; D008955:Models, Cardiovascular; D010957:Plasmids / Q000737:chemistry; D011073:Polyamines / Q000737:chemistry; D051381:Rats
chemical_list: D000974:Antibodies, Antinuclear; D002317:Cardiovascular Agents; D004164:Diphosphonates; D008823:Micelles; D011073:Polyamines; D049452:Green Fluorescent Proteins; C063994:polyallylamine; D004247:DNA
keywords: gene delivery; micelles; plasmid DNA; polyallylamine bisphosphonate; stent