Changes in cell spreading and cytoskeletal organization are induced by adhesion to a fibronectin-fibrin matrix

Plasma fibronectin (pFN) cross-linked to fibrin during the injury response provides a provisional matrix required for cells to begin tissue repair. Using a synthetic matrix of pFN and fibrin as a substrate for cell adhesion and spreading, we have determined that pFN covalently cross-linked to fibrin into a complex multimer is functionally distinct from pFN immobilized onto a plastic surface. NIH-3T3 cells on a FN-fibrin matrix reach 50% of the maximal cell area of cells spread on FN-coated plastic. They neither attach nor spread on cross-linked fibrin alone. Cells on pFN-fibrin matricesform few prominent stress fibers and exhibit clear differences in membrane ruffling and filopodial extension when stained with rhodamine- labeled phalloidin. Interestingly, these differences are enhanced by upregulation of protein kinase C. These data suggest that cell-FN interactions can be modified by the molecular context of the protein within the extracellular matrix resulting in distinct cell morphology and cytoskeletal organization.