Chronic wound healing states are often associated with aging, and despite the increased number of aged patients with nonhealing wounds, controversy still exists concerning the effects of age on wound repair. Our previous work showed that in both venous ulcers in humans and acute wounds in aged animals, fibronectin, an early component in granulation tissue, is deficient compared to normal skin and acute wounds in healthy young animals, respectively. In the present study, we have determined the protease responsible for fibronectin degradation by analyzing tissue taken from the margins of chronic venous ulcers and standardized acute cutaneous wounds collected from a large cohort of" Health status"-defined aged human subjects (screened as per the SENIEUR protocol). When tissue samples were subjected to fibronectin zymography, the main protease involved in the breakdown of fibronectin in both venous ulcers and acute wounds of elderly subjects was found to be a serine protease with a molecular weight of approximately 30 kd. This protease was identified as neutrophil elastase by immunoblotting. In tissue biopsies, elastase was localized to granulocytes by immunocytochemical techniques and shown to be present in greater quantities in venous ulcers and Day-7 and-14 healing acute wounds of healthy aged subjects relative to those of young subjects. The highest quantities were found in acute wounds of elderly women. Our results suggest that the process of aging in healthy human subjects is associated with an up-regulation of elastase during acute wound healing and that an abnormality in down-regulation of this protease could be partially responsible for the transition to chronic wound healing states in the aged.