Fibroblast Growth Factor 2 Enhances Early Stages of in Vitro Endothelial Repair by Microfilament Bundle Reorganization and Cell Elongation

Abstract

As endothelial cells convert from quiescent to migrating cells over 8 h along a wound edge, actin microfilaments undergo well-defined sequential changes characterized by an initial random distribution followed by a parallel and then a perpendicular orientation of microfilaments with respect to the wound edge. The latter is associated with subsequent cell migration. We tested the hypothesis that fibroblast growth factor 2 (FGF-2) can enhance the very early stages of wound repair even prior to migration and that FGF-2 enhancement of wound repair is associated with changes in the endothelial actin cytoskeleton. Using an in vitro two-sided wound model, the addition of FGF-2 at the time of wounding enhanced the extent of wound closure over 8 h. Treatment with FGF-2 was associated with significantly longer cells along the wound edge at 4 and 8 h after wounding. When treated with increasing concentrations of neutralizing FGF-2 antibody, the extent of wound closure decreased over 8 h and was associated with a decrease in cell length along the wound edge. Actin microfilaments were local ized using rhodamine phalloidin and viewed using laser confocal microscopy. At 4 h after wounding, FGF-2 treatment was associated with significantly more cells along the wound expressing perpendicular microfilaments compared to untreated cells, which suggested a more rapid transition of parallel to perpendicular microfilament distribution. Thus, FGF-2 affects the very early stages of wound repair prior to migration by enhancing wound closure due to the early appearance of perpendicular microfilaments and lengthening of cells along the wound edge.