[Inhibiting scar formation in rat cutaneous wounds by blocking TGF-beta signaling].
OBJECTIVE
TGF-beta plays a key role in wound scarring. This study explored the possibility of using gene therapy to inhibit wound scarring by blocking TGF-beta signaling.


METHODS
In vitro, human normal dermal fibroblasts were infected with recombinant adenoviruses of truncated TGF-beta receptor II (tTGF-betaRII, 100 pfu/cell) and beta-galactosidase (beta-gal, 100 pfu/cell), and their effects on regulating TGF-beta1 gene expression were analyzed by Northern blot. For gene therapy, beta-gal and tTGF-betaRII viruses (1 x 10(9) pfu)were injected intradermally at left and right side of dorsal skin of newborn Sprague-Dawley rats (n = 15) respectively. A full-thickness incisional wound (0.5 cm long) was created at the injection sites of each rat 2 days post-injection. An incisional wound was similarly created in the middle part of the dorsal skin of tTGF-betaRII transgenic mice (n = 5) and control mice (n = 5). Wound tissues of rats and mice were harvested at various time points post-wounding for histological and immunohistochemical analysis. Scar area in tissue section was measured by Image-Pro Plus software.


RESULTS
Over-expression of tTGF-betaRII markedly reduced TGF-beta1 gene expression in dermal fibroblasts. Adenovirus mediated gene expression in skin reached a peak level 2 - 3 days post-injection, and decreased gradually at 5 - 7 days. Two weeks post-wounding, histology and quantitative analysis demonstrated that relative scar area in the wounds of transgenic mice and control mice were 136,969.8 +/- 66,339 and 474,641.6 +/- 227,396 respectively, the scar area of transgenic wounds was 29 percent of control area (P < 0.05). In all rats, wounds transfected with tTGF-betaRII gene healed with much less scarring (relative scar area 128,311.2 +/- 36,764.6) than control wounds (251,189.1 +/- 62,544.7) of the same rat, with a 45% reduction of scar area in average (P < 0.001). In addition, the tTGF-betaRII expression also decreased inflammation and TGF-beta1 production in treated wounds, and promoted the repair of panniculus muscle in treated wounds.


CONCLUSIONS
Adenovirus mediated over-expression of tTGF-betaRII can block TGF-beta signaling and inhibit wound scarring, and thus can serve as a gene therapy strategy to control wound scarring.
Liu
Wei
W
Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Second Medical University, Shanghai 200011, China.
Chua
Chek Hau
CH
Wu
Xiao-li
XL
Wang
Dan-ru
DR
Yin
De-min
DM
Cui
Lei
L
Cao
Yi-lin
YL
Longaker
Michael T
MT
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002921:Cicatrix / Q000628:therapy*; D004195:Disease Models, Animal; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D051381:Rats; D017207:Rats, Sprague-Dawley; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics / Q000627:therapeutic use*; D015398:Signal Transduction / Q000502:physiology; D012871:Skin Diseases / Q000378:metabolism / Q000628:therapy*; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000378:metabolism; D053773:Transforming Growth Factor beta1; D014945:Wound Healing / Q000502:physiology*
D018125:Receptors, Transforming Growth Factor beta; C508960:TGFB1 protein, human; C508961:Tgfb1 protein, mouse; C508962:Tgfb1 protein, rat; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1