This study used musculoskeletal modelling to explore the relationship between cycling conditions (power output and cadence) and muscle activation and metabolic power. We hypothesized that the cadence that minimized the simulated average active muscle volume would be higher than the cadence that minimized the simulated metabolic power. We validated the simulation by comparing the predicted muscle activation and fascicle velocities with experimental electromyography and ultrasound images. We found strong correlations for averaged muscle activations and moderate to good correlations for fascicle dynamics. These correlations tended to weaken when analyzed at the individual participant level. Our study revealed a curvilinear relationship between the average active muscle volume and cadence, with the minimum active volume being aligned to the self-selected cadence. The simulated metabolic power was consistent with previous results and was minimized at lower cadences than that which minimized active muscle volume across power outputs. Although there are some limitations to the musculoskeletal modelling approach, the findings suggest that minimizing active muscle volume may be a more important factor than minimizing metabolic power for self-selected cycling cadence preferences. Further research is warranted to explore the potential of an active muscle volume-based objective function for control schemes across a wider range of cycling conditions.