Effect of transfection time on the survival of epigastric skin flaps pretreated with adenovirus encoding the VEGF gene.
An experimental study was conducted to investigate the effect of time of adenovirus-mediated vascular endothelial growth factor (VEGF) gene therapy on the viability of epigastric skin flaps. Eighty-four male Sprague-Dawley rats were used. Skin flaps measuring 8 x 8 cm were marked on the ventral abdominal wall. The upper border of the flap was 1 cm above the costal margin, and the lower border was at the pubis and the inguinal fold. The lateral borders of the flap corresponded to the location of the distinct conversion of the thin ventral skin to the thick dorsal skin. Seven sites in the predicted area of necrosis on the outlined skin flaps were chosen for subdermal injections. All injections were administered by an individual who was blinded to the different treatment groups. The rats received either saline (control group I, N = 28) or adenovirus encoding green fluorescent protein (Ad-GFP; group II, N = 28) or Ad-VEGF (group III, N = 28). The epigastric island skin flaps based solely on the right inferior epigastric vessels were elevated either on the same day of injection (day 0 = 12 hours after transfection, N = 7) or on day 3 (N = 7), day 7 (N = 7), or day 14 (N = 7) after subdermal gene therapy. Flaps were sutured back to their native configuration. Flap viability was evaluated on day 7 after surgery. Sections of the flaps were examined histologically after undergoing hematoxylin-eosin staining. There was a significant reduction in mean percentage of necrotic flap area by 56%, 67%, 70%, and 54% in flaps transfected with Ad-VEGF, 12 hours, 3 days, 7 days, and 14 days before flap elevation, respectively ( < 0.05). There was no evidence that the mean percentage of skin necrosis in the Ad-GFP group was different than in the control group ( = 0.26). There was evidence of mild inflammation in flaps pretreated with Ad-GFP and Ad-VEGF compared with the control group. The authors demonstrated that adenovirus-mediated gene therapy of the abdominal skin after subdermal injections was technically feasible. This was demonstrated by the visualization of GFP expression in control experiments using a fluorescence microscope. In this study, adenovirus-mediated VEGF gene therapy promoted epigastric flap survival, which was not related to the time of transfection. These findings raise the possibility that pretreatment with VEGF gene therapy using an adenovirus vector may be applicable in patients at risk for plastic surgery.
Department of Plastic and Reconstructive Surgery, The Cleveland Clinic Foundation, OH 44195, USA.
Raffi
R
Gurunluoglu
Kagan
K
Ozer
Blazenka
B
Skugor
Przemyslaw
P
Lubiatowski
Kevin
K
Carnevale
Maria
M
Siemionow
D034861:Abdominal Wall; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016228:Endothelial Growth Factors / Q000235:genetics*; D019074:Epigastric Arteries; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D008164:Luminescent Proteins; D008222:Lymphokines / Q000235:genetics*; D008297:Male; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D009336:Necrosis; D018919:Neovascularization, Physiologic / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013524:Surgical Flaps / Q000098:blood supply* / Q000473:pathology; D013997:Time Factors; D014022:Tissue Survival; D014162:Transfection / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016228:Endothelial Growth Factors; D007202:Indicators and Reagents; D036341:Intercellular Signaling Peptides and Proteins; D008164:Luminescent Proteins; D008222:Lymphokines; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D049452:Green Fluorescent Proteins