Our previous experimental work with tissue plasminogen activator (TPA) suggested the possibility of the clearance of vitreous hemorrhage by repetitive injections of low-dose TPA. We therefore investigated in rabbits the effect of both repeated injections of TPA and the change of the integrity of the vitreous body on the clearance of vitreous hemorrhage. Vitreous hemorrhage was produced by intravitreal injection of 0.05 ml of autologous whole blood in the pigmented rabbit eyes with intact vitreous or gas-compressed vitreous. Three intravitreal injections of 3-g TPA (total dose of 9 microgram), separated by 7-day intervals, were performed. The endpoint for vitreous hemorrhage clearance was defined as clear visualization of the posterior central retina of the rabbits. Regardless of whether gas compression vitrectomy was performed, repeated injections of low-dose TPA resulted in rapid clearance of fresh vitreous hemorrhage in approximately two to three weeks after the last TPA injection. No evidence of retinal toxicity was seen in all experimental groups. Repetitive injections of low-dose TPA may be effective in the treatment of fresh vitreous hemorrhage.