In vitro analysis of transforming growth factor-beta1 inhibition in novel transgenic SBE-luciferase mice.
BACKGROUND
Transforming growth factor beta1 (TGF-beta1) expression correlates with scarring. A novel transgenic mouse model with a Smad2/3-responsive luciferase reporter construct (SBE-luc) has been developed. We hypothesized that bioluminescence in SBE-luc dermal fibroblasts could be measured to assess TGF-beta1 inhibition.


METHODS
Cultured dermal fibroblasts from SBE-luc mice were treated simultaneously with TGF-beta1 and increasing doses of either neutralizing antibody to TGF-beta (NA-TGFbeta) or SB-431542, a novel TGF-beta receptor kinase inhibitor. Fibroblasts were measured for luciferase activity. SBE-luc fibroblasts underwent Western blot analysis for collagen type I production.


RESULTS
TGF-beta1 produced maximal luciferase activity in SBE-luc fibroblasts at 0.1 ng/mL (P < 0.05). NA-TGFbeta and SB-431542 inhibited luciferase activity in a dose-dependent fashion, with complete inhibition achieved by 0.1 microg/mL and 1 microM, respectively (P < 0.05). NA-TGFbeta and SB-431542 inhibited collagen type I production.


CONCLUSIONS
Our in vitro results provide validation for further in vivo real-time imaging studies using the SBE-luc mouse as a novel wound-healing model.
Satterwhite
Thomas S
TS
Section of Plastic Surgery, Department of Veterans Affairs and Division of Plastic Surgery, Stanford University Medical Center, Stanford, CA 94305, USA.
Chong
Alphonsus K
AK
Luo
Jian
J
Pham
Hung
H
Costa
Melinda
M
Longaker
Michael T
MT
Wyss-Coray
Tony
T
Chang
James
J
D000818:Animals; D001549:Benzamides / Q000494:pharmacology*; D002478:Cells, Cultured; D002921:Cicatrix / Q000209:etiology; D004149:Dioxoles / Q000494:pharmacology*; D005347:Fibroblasts / Q000502:physiology*; D008156:Luciferases; D051379:Mice; D008822:Mice, Transgenic; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors* / Q000502:physiology*; D014945:Wound Healing
C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D004149:Dioxoles; D053773:Transforming Growth Factor beta1; D008156:Luciferases