Fibronectin plays an important role in wound healing and cell differentiation. The present study measures fibronectin levels in aqueous humor from injured corneas. In response to transcorneal freezing treatments, a thickened scar-like matrix is deposited on the posterior surface of rabbit corneas. This posterior collagenous layer (PCL) is visible grossly and has a uniform diameter. Aqueous humor from eyes with established posterior corneal scarring has significantly less fibronectin compared with control eye aqueous humor. Tissue culture experiments show that corneal endothelial cells from control eyes and PCL-associated cells from injured eyes produce similar amounts of fibronectin. The marked decline in fibronectin levels in aqueous humor in scarred corneas supports the hypothesis that fibronectin plays a role in the pathological events which result from corneal injury.