Inhibiting scar formation in rat wounds by adenovirus-mediated overexpression of truncated TGF-beta receptor II.
The purpose of this study was to explore the possibility of inhibiting wound scarring by blocking TGFbeta signaling of wound cells by means of a gene therapy approach. Normal dermal fibroblasts were infected in vitro either with recombinant adenovirus encoding a truncated TGFbeta receptor II (Ad-tTGF-betaRII) or with [beta]-galactosidase adenovirus (Ad-beta-gal). TGF-beta1 gene expression in infected fibroblasts was analyzed by Northern blot. In vivo, 1x10(9) plaque-forming units of Ad-tTGF-betaRII were intradermally injected into the dorsal skin of 10-day-old newborn Sprague-Dawley rats (n = 10). For gene therapy, 1x10(9) plaque-forming units of Ad-tTGF-betaRII viruses were injected intradermally at the right side dorsal skin of another set of same aged Sprague-Dawley rats as the experimental group (n = 15). In the control group, 1x10(9) plaque-forming units of Ad-beta-gal (n = 11) or the same volume of saline (n = 4) was injected at the left side skin of the same rats. A 5-mm-long full-thickness incisional wound was created at the injection sites of each rat 2 days after injection. Wound tissues were harvested at day 3 (n = 2), day 7 (n = 2), and day 14 (n = 11) after wounding for histological analysis. Scar area of wound tissues harvested at day 14 was quantitatively analyzed. The results showed that TGF-beta1 gene expression was markedly down-regulated in Ad-tTGF-betaRII infected fibroblasts compared with Ad-beta-gal infected cells. In vivo, adenovirus-mediated transgene expression in rat skin reached a peak level at day 2 after injection and the expression gradually decreased afterward. Inhibited inflammatory reaction was also observed in the treated wounds with significantly reduced inflammatory cells (p < 0.05). Moreover, in all 11 rats, the experimental wound at day 14 had much less scarring than its control wound of the same rat, with an average of 49 percent reduction of the scar area (p < 0.05). Furthermore, more panniculus muscles were repaired in the experimental wounds (nine of 11) than in the control wounds (two of 11) (p < 0.05). These results indicate that gene therapy by targeting wound TGF-beta can effectively inhibit wound scarring and may potentially be applied to clinical scar treatment.
Liu
Wei
W
Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Second Medical University, Shanghai Institute of Plastic and Reconstructive Surgery, Shanghai Tissue Engineering Center, P.R. China. liuwei_2000@yahoo.com
Chua
Chekhau
C
Wu
Xiaoli
X
Wang
Danru
D
Ying
Demin
D
Cui
Lei
L
Cao
Yilin
Y
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D015152:Blotting, Northern; D002478:Cells, Cultured; D002921:Cicatrix / Q000378:metabolism* / Q000517:prevention & control*; D016895:Culture Media, Serum-Free; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D011509:Proteoglycans / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014945:Wound Healing / Q000502:physiology*
D016895:Culture Media, Serum-Free; D011509:Proteoglycans; D018125:Receptors, Transforming Growth Factor beta; C071736:betaglycan