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- Title
Ex vivo transfer of Smad7 decreases damage to the corneal endothelium after penetrating keratoplasty.
- Authors
Toshinari Funaki; Nobuyuki Ebihara; Akira Murakami; Atsuhito Nakao
- Abstract
Abstract Purpose To determine whether transient gene transfer and expression of the intracellular antagonist of transforming growth factor β (TGF-β), Smad7, to corneal endothelial cells decreases corneal endothelial cell damage after penetrating keratoplasty in a rabbit model. Methods Rabbit corneas were transfected ex vivo with replication-deficient adenoviruses encoding Flagtagged Smad7, Flag-tagged Smad3, or LacZ (termed AdCMV-Smad7, AdCMV-Smad3, AdCMV-LacZ) and then transplanted to normal rabbits. Expression of the exogenous Smads and phosphorylation of endogenous Smad2 in the transplanted corneal endothelium were examined by immunoblotting and immunohistochemistry with anti-Flag or anti-phosphorylated Smad2 antibodies. Cellular density and morphological changes in the corneal endothelium of the transplanted cornea were evaluated by scanning electron microscopy after transplantation of the Smad-transfected corneas. Results Transplanted AdCMV-Smad7-transfected corneas significantly inhibited the decrease in cellular density and accelerated wound healing at the host-graft junction when compared with transplanted AdCMV-LacZ-transfected corneas. Transplanted AdCMV-Smad3-transfected corneas showed decreased cellular density and delayed wound healing at the host-graft junction. Conclusions Ex vivo gene transfer of Smad7 to corneal endothelial cells inhibits the decrease in cellular density and accelerates wound healing after penetrating keratoplasty in rabbits. Thus, modulation of Smad7 expression in corneal endothelial cells may decrease corneal endothelial cell damage after penetrating keratoplasty.
- Subjects
CORNEA diseases; ENDOTHELIUM; CORNEA surgery; WOUND healing; GENETIC transformation; ELECTRON microscopy; CYTOKINES
- Publication
Japanese Journal of Ophthalmology, 2008, Vol 52, Issue 3, p204
- ISSN
0021-5155
- Publication type
Article
- DOI
10.1007/s10384-007-0526-2