Cytoskeleton in TFG-β- and bFGF-modulated endothelial monolayer repair

Abstract

Endothelial cell proliferation and migration in vitro is depressed by transforming growth factor beta (TFG-β) and enhanced by basic fibroblast growth factor (bFGF) treatment. This study examines interactions between cytoskeletal changes and cell proliferation in regenerating endothelial monolayers treated with bFGF, TFG-β, and both factors. As previously described by others, monolayer regeneration is enhanced by bFGF and reduced by TFG-β. Endothelial cell morphology is altered by TFG-β treatment. Cells lose their cobblestone appearance and assume a pleomorphic shape. Actin microfilament staining is modified in both intact and regenerating TFG-β-treated monolayers as well. There is a loss of dense peripheral band staining and an enhancement in staining intensity of cytoplasmic stress fibers. No such alterations are seen in bFGF-treated cultures. Cell proliferation at the wound edge, as indicated by bromodeoxyuridine incorporation, is inhibited by TGF-β. Although monolayer repair is modulated by growth factor treatment, centrosome reorientation and microtubule staining patterns are not altered by either factor. Thus these factors appear to have effects on a mechanism(s) other than centrosome reorientation which may be involved in repair of denuded endothelial monolayers.