Background. Many sports and physical activities can result in lower limb injures. Pedaling is an effective exercise for lower extremity rehabilitation, but incorrect technique may cause further damage. To some extent, previous experiments have been susceptible to bias in the sample recruited for the study. Alternatively, methods used to simulation activities can enable parametric studies without the influence of noise. In addition, models can facilitate the study of all muscles in the absence of the effects of fatigue. This study investigated the effects of crank length on muscle behavior during pedaling. Methods. Six muscles (soleus, tibialis anterior, vastus medialis, vastus lateralis, gastrocnemius, and rectus femoris), divided into three groups (ankle muscle group, knee muscle group, and biarticular muscle group), were examined under three cycling crank lengths (100 mm, 125 mm, and 150 mm) in the present study. In addition, the relationship between crank length and muscle biological force was analyzed with the AnyBody Modeling System™, a human simulation modeling software based on the Hill-type model. Findings. Based on inverse kinematic analysis, the results indicate that muscle activity and muscle force decrease in varying degrees with increases in crank length. The maximum and minimum muscular forces were attained in the tibialis anterior and vastus lateralis, respectively. Interpretation. Studying the relationship between muscle and joint behavior with crank length can help rehabilitation and treating joint disorders. This study provides the pedal length distribution areas for patients in the early stages of rehabilitation.