Abstract Background Cycling is commonly prescribed for knee osteoarthritis, but previous literature on biomechanics during cycling and the effects of acute intervention on osteoarthritis patients does not exist. Due to their altered knee kinematics, osteoarthritis patients may be at greater risk of osteoarthritis progression or other knee injuries during cycling. This study investigated the effects of reduced foot progression (toe-in) angles on knee joint biomechanics in subjects with medial compartment knee osteoarthritis. Methods Thirteen osteoarthritis and 11 healthy subjects participated in this study. A motion analysis system and custom instrumented pedal was used to collect 5 pedal cycles of kinematic and kinetic data in 1 neutral and 2 toe-in conditions (5 and 10) at 60 RPM and 80 W. Findings For peak knee adduction angle, there was a 61% (2.7) and a 73% (3.2) decrease in the 5 and 10 toe-in conditions compared to neutral in the osteoarthritis group and a 77% (1.7) and 109% (2.4) decrease in the healthy group for the 5 and 10 conditions, respectively. This finding was not accompanied by a decrease in pain or peak knee abduction moment. A simple linear regression showed a positive correlation between Kelgren-Lawrence score and both peak knee adduction angle and abduction moment. Interpretation For individuals who cycle with increased knee adduction angles, decreasing the foot progression angle may be beneficial for reducing the risk of overuse knee injuries during cycling by resulting in a frontal plane knee alignment closer to a neutral position.