Optimization of ex vivo pressure mediated delivery of antisense oligodeoxynucleotides to ICAM-1 reduces reperfusion injury in rat cardiac allografts.
BACKGROUND
Our purpose was to optimize hyperbaric pressure as a vector for ex vivo transfection of antisense oligodeoxynucleotides (AS-ODN) to intercellular adhesion molecule-1 to limit reperfusion injury (RI) in cardiac allografts. We investigated the effects of increased pressure, incubation time, and AS-ODN concentrations on transfection efficiency and toxicity.


RESULTS
PVG (RT1c) donor hearts were heterotopically transplanted to ACI (RT1a) recipients. Donor hearts were harvested and the various groups were treated at: (1) different pressure (1-9 atm) for 45 min with 80 micromol/liter AS-ODN; (2) different incubation times (15 min to 6 hr) at 5 atm with 80 micromol/liter AS-ODN; 3) different AS-ODN concentrations (80-240 micromol/liter) at 5 atm for 45 min. Hearts were procured 24 or 72 hr after transplantation. Transfection efficiency was determined with fluorescein-labeled AS-ODN. The degree of RI was determined with biochemical and histological analysis. Increasing pressure from ambient (1 atm) pressure to pressures as high as 9 atm leads to a increase in transfection efficiency from 1.7+/-.5 to 62+/-3.9% and a reduction in RI. Increased incubation time up to 45 min increased transfection efficiency and reduced RI, but longer incubation times induced significant toxicity to the allograft. Increased AS-ODN concentrations improved transfection and reduced RI.


CONCLUSIONS
Hyperbaric pressure is a safe and effective vector for the ex vivo delivery of AS-ICAM-1-ODN to rodent cardiac allografts and results in a reduction in reperfusion injury.
Feeley
B T
BT
Department of Cardiothroacic Surgery, Stanford University School of Medicine, California 94305, USA.
Poston
R S
RS
Park
A K
AK
Ennen
M P
MP
Hoyt
E G
EG
Vriens
P W
PW
Robbins
R C
RC
D000818:Animals; D015316:Genetic Therapy; D016027:Heart Transplantation; D006931:Hyperbaric Oxygenation; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics*; D008297:Male; D016376:Oligonucleotides, Antisense / Q000627:therapeutic use / Q000633:toxicity*; D011312:Pressure; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011912:Rats, Inbred ACI; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000517:prevention & control* / Q000628:therapy*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors; D014162:Transfection
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D018799:Intercellular Adhesion Molecule-1