The vitreous is a gel-like connective tissue that undergoes liquefaction during aging and pathological processes. We isolated and characterized a degradative enzyme from the vitreous of different species and identified it to be matrix metalloproteinase-2 (MMP-2). This enzyme was found in a latent form and may be associated with endogenous inhibitors. Vitreous isolated from both non-diabetic and diabetic patients contained MMP-2 in the same concentrations. However, the diabetic samples had an additional gelatinase activity at 92 kDa which may be associated with a compromised vasculature. These results suggest that the normal human vitreous contains an endogenous MMP and the appearance of an additional activity is associated with pathologic conditions. Curr. Eye Res. 13: 639–647, 1994.