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Background:Q-Factor (QF), or the inter-pedal width, in cycling is similar to step-width in gait. Although increased step-width has been shown to reduce peak knee abduction moment (KAbM), no studies have examined the biomechanical effects of increased QF in cycling at different work-rates in healthy participants.Methods:A total of 16 healthy participants (8 males, 8 females, age:22.4 ± 2.6 years, body mass index:22.78 ± 1.43 kg/m2, mean ± SD) partic-ipated. A motion capture system and customized instrumented pedals were used to collect 3-dimensional kinematic (240 Hz) and pedal reaction force (PRF) (1200 Hz) data in 12 testing conditions:4 QF conditions—Q1 (15.0 cm), Q2 (19.2 cm), Q3 (23.4 cm), and Q4 (27.6 cm)—under 3 workrate conditions—80 watts (W), 120 W, and 160 W. A 3 × 4 (QF × workrate) repeated measures of analysis of variance were performed to analyze differences among conditions (p<0.05).Results:Increased QF increased peak KAbM by 47％, 56％, and 56％from Q1 to Q4 at each respective workrate. Mediolateral PRF increased from Q1 to Q4 at each respective workrate. Frontal-plane knee angle and range of motion decreased with increased QF. No changes were observed for peak vertical PRF, knee extension moment, sagittal plane peak knee joint angles, or range of motion.Conclusion:Increased QF increased peak KAbM, suggesting increased medial compartment loading of the knee. QF modulation may influence frontal-plane joint loading when using stationary cycling for exercise or rehabilitation purposes.