Rotator cuff tears frequently occur and can lead to pain and decreased shoulder function. Repair of the torn tendon back to bone is often successful in relieving pain, but failure of the repair commonly occurs. Post-operative activity level is an important treatment component that has received minimal attention for the shoulder, but may have the potential to enhance tendon to bone healing. The objective of this study was to investigate the effect of short and long durations of various activity levels on the healing supraspinatus tendon to bone insertion site. Rotator cuff tears were surgically created in Sprague–Dawley rats by detaching the supraspinatus tendon from its insertion on the humerus and these tears were immediately repaired back to the insertion site. The post-operative activity level was controlled through shoulder immobilization (IM), cage activity (CA), or moderate exercise (EX) for durations of 4 or 16 weeks. The healing tissue was evaluated utilizing biomechanical testing and a quantitative polarized light microscopy method. We found that activity level had no effect on the elastic properties (stiffness, modulus) of the insertion site at four weeks post injury and repair, and a decreased activity level had a positive effect on these properties at 16 weeks . Furthermore, a decreased activity level had the greatest positive effect on these properties over time . The angular deviation of the collagen, a measure of disorganization, was decreased with a decrease in activity level at 4 weeks , but was similar between groups at 16 weeks . It appears from this study that decreasing the activity level by immobilizing the shoulder improves tendon to bone healing, which progresses by first increasing the organization of the collagen and then increasing the mechanical properties. Future studies in this area will investigate the effect of passive motion and remobilization on both tendon to bone healing and shoulder function.