J Mohay1, B J McLaughlin. 1. Department of Ophthalmology & Visual Sciences, Kentucky Lions Eye Research Institute, University of Lousville School of Medicine 40292, USA.
Abstract
BACKGROUND: The aim of the present study was to compare the morphology, proliferative activity and cytoskeletal organization of bovine corneal endothelial cells during wound healing under normal and mitotically inhibited conditions. METHODS: Cell cultures were grown to confluency and incubated with the mitotic inhibitor 5-fluorouracil (5-FU; 2.5 micrograms/ml) followed by a touch wound. Control cultures were maintained without 5-FU. Mitotic activity, F-actin, vinculin, vimentin and connexin 43 localization were evaluated before, during and after wound closure. RESULTS: 5-FU inhibited irreversibly the mitotic activity of corneal endothelial cells during the whole wound healing process. In the presence of 5-FU, a high degree of polymegathism and delay in actin and vinculin redistribution to the cell borders after wound closure was observed. Vimentin and connexin 43 immunolabeling revealed only slight differences between 5-FU-treated and control cultures. CONCLUSIONS: Significant changes in cell geometry and cytoskeletal organization in the amitotic corneal endothelium became manifested only after wounding. These changes may influence cell-cell and cell-matrix interactions as well as functional restoration of the monolayer after wound closure.
BACKGROUND: The aim of the present study was to compare the morphology, proliferative activity and cytoskeletal organization of bovine corneal endothelial cells during wound healing under normal and mitotically inhibited conditions. METHODS: Cell cultures were grown to confluency and incubated with the mitotic inhibitor 5-fluorouracil (5-FU; 2.5 micrograms/ml) followed by a touch wound. Control cultures were maintained without 5-FU. Mitotic activity, F-actin, vinculin, vimentin and connexin 43 localization were evaluated before, during and after wound closure. RESULTS:5-FU inhibited irreversibly the mitotic activity of corneal endothelial cells during the whole wound healing process. In the presence of 5-FU, a high degree of polymegathism and delay in actin and vinculin redistribution to the cell borders after wound closure was observed. Vimentin and connexin 43 immunolabeling revealed only slight differences between 5-FU-treated and control cultures. CONCLUSIONS: Significant changes in cell geometry and cytoskeletal organization in the amitotic corneal endothelium became manifested only after wounding. These changes may influence cell-cell and cell-matrix interactions as well as functional restoration of the monolayer after wound closure.