Lipofuscin accumulation in cultured rat myocardial cells is considered an index of intra-lysosomal oxidative reactions and was registered by autofluorescence measurements. Lipofuscinogenesis in secondary lysosomes is thought to be a consequence of Fenton reactions, and is much enhanced by oxidative stress obtained by culturing the cells in an atmosphere containing 40% oxygen. The influence of the synthetic antioxidant indenoindole (DHII), as compared to control cells, was a dose-responsive depression of lipofuscinogenesis to a degree of 19% and 17% with 20 μM DHII and to 25% and 23% with 40 μM DHII after 7 and 14 days in culture, respectively. This demonstrates a significant quenching of oxidative stress and suggests the therapeutic value of DHII and related antioxidants in protecting against oxygen radical-related diseases. It is also suggested that neonatal cardiac myocytes in culture are a suitable model system for the evaluation of oxygen radical-induced myocardial damage.