In Vitro Analysis of Transforming Growth Factor-β1 Inhibition in Novel Transgenic SBE-Luciferase Mice


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Abstract

Background:Transforming growth factor β1 (TGF-β1) 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-β1 inhibition.Materials and Methods:Cultured dermal fibroblasts from SBE-luc mice were treated simultaneously with TGF-β1 and increasing doses of either neutralizing antibody to TGF-β (NA-TGFβ) or SB-431542, a novel TGF-β receptor kinase inhibitor. Fibroblasts were measured for luciferase activity. SBE-luc fibroblasts underwent Western blot analysis for collagen type I production.Results:TGF-β1 produced maximal luciferase activity in SBE-luc fibroblasts at 0.1 ng/mL (P < 0.05). NA-TGFβ and SB-431542 inhibited luciferase activity in a dose-dependent fashion, with complete inhibition achieved by 0.1 μg/mL and 1 μM, respectively (P < 0.05). NA-TGFβ 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.

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