Keratoconus is an ocular disorder in which the central cornea becomes thin, conical and frequently scarred. We are exploring the possibility that this condition is induced and maintained by proteases that exist in the corneal matrix in an activated form. In this study, the activities of the proteases secreted in vitro and in vivo by keratocytes of normal, clear keratoconic, scarred keratoconic and traumatically scarred corneas have been compared and partially characterised. Data obtained by assaying acyl transferase activity showed that the matrix metalloproteinases account for a minimum of 95% of the total protease secreted by cultured keratocytes. Their summated specific activity was consistently and significantly higher in the culture medium of keratoconic keratocytes than in the medium of other keratocyte cultures. Analysis of the individual protease activities secreted by these corneal keratocytes in vitro and in vivo by SDS-gelatin polyacrylamide gel electrophoresis showed that a gelatinase of molecular weight 65,000 is the major protease secreted by normal keratocytes. Whereas clear keratoconic and traumatically scarred corneal keratocytes secrete an additional activity of molecular weight 61,000, scarred keratoconic corneal keratocytes generally produced little or none of this gelatinase activity. Both activities may be ascribed to gelatinase A, and although the 61,000 molecular weight form may be a significant feature of keratoconus, neither appears to be active as secreted.