Apoptosis in Cultured Corneal Stromal Fibroblasts of Keratoconus Treated with Fas-Stimulating Antibody

Choi, Sang Il; Cho, Eun Young; Kim, Woo Jung

Abstract

PURPOSE: Fas/Fas ligand system plays an important role in modulating keratocyte apoptosis induced by mechanical corneal epithelial injury. It is also hypothesized that keratocyte apoptosis could be involved in the pathogenesis of keratoconus. The purpose of this study is to evaluate apoptosis in the first passage cultured corneal stromal fibroblasts of normal and keratoconus cornea treated with Fas-stimulating antibody.
METHODS
Apoptotic cell death was evaluated by trypan blue exclusion assay in the first passage cultured corneal stromal fibroblasts of keratoconus treated with Fas-stimulating antibody. The experiment was performed in comparison with normal corneal stromal fibroblasts as a control.
RESULTS
Apoptotic cells were found in the first passage cultured stromal fibroblasts of normal and keratoconus cornea treated with Fas-stimulating antibody. Apoptotic reaction of cultured stromal fibroblasts of keratoconus cornea treated with Fas-stimulating antibody was stronger than that of normal cultured corneal stromal fibroblasts treated with Fas-stimulating antibody. Apoptosis did not occur in cultured stromal fibroblasts of normal and keratoconus cornea treated with normal mouse control IgM. In Hoechst staining of cell suspension including apoptotic cells, characteristic findings such as cell shrinkage and chromatin condensation were observed.
CONCLUSIONS
This study showed the differential features of cell death in cultured corneal stromal fibroblasts of keratoconus compared with normal control. Thus, keratocyte apoptosis induced by Fas/Fas ligand system could be an important factor in the pathogenesis of keratoconus.