The aim of the study was to examine whether self-selected walking speed during downhill treadmill walking by older adults would result in muscle injury and changes in physiological responses during level walking. Twenty-six participants (age: 67 ± 4 yrs; height: 1.69 ± 0.09 m; body mass: 74.9 ± 13.1kg) were assigned to level (n = 11, 30 min, 0%) or downhill walking (n=15, 30 min, -10%) at a self-selected walking speed. Self-selected walking speed and exercise intensity were similar for both groups (level: 4.2±0.4 km·hr-1, 42±6% VO2max; downhill: 4.6±0.6 km·hr-1, 44±15% VO2max). After 48-hours, downhill walking had reduced maximal voluntary isometric force of the m. quadriceps femoris (-15%, P<0.001), indicative of muscle injury, but no changes were observed for walking economy, minute ventilation, heart rate and respiratory exchange ratio during level walking. For older adults, downhill walking at a selfselected walking speed causes muscle injury without any detrimental effect on walking economy. Regular downhill walking at a self-selected walking speed by older adults is an eccentric endurance activity that may have the potential to improve cardiovascular fitness and muscle strength.