CORNEAL ENDOTHELIAL WOUND REPAIR IN NORMAL AND MITOTICALLY INHIBITED CULTURES

Background: The aim of the present study was to compare the morphology , proliferative activity and cytoskeletal organization of bovine corne al 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 mu g/ml) followed by a touch wound. Control cultures were maintained wit hout 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 prese nce of 5-FU, a high degree of polymegathism and delay in actin and vin culin redistribution to the cell borders after wound closure was obser ved. Vimentin and connexin 43 immunolabeling revealed only slight diff erences between 5-FU-treated and control cultures. Conclusion: Signifi cant changes in cell geometry and cytoskeletal or anization in the ami totic 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.