Objective. To investigate the in vivo cartilage contact biomechanics of the tibiofemoral joint following anterior cruciate ligament (ACL) injury.Methods. Eight patients with an isolated ACL injury in 1 knee, with the contralateral side intact, participated in the study. Both knees were imaged using a specific magnetic resonance sequence to create 3-dimensional models of knee bone and cartilage. Next, each patient performed a lunge motion from 0°to 90°of flexion as images were recorded with a dual fluoroscopic system. The three-dimensional knee models and fluoroscopic images were used to reproduce the in vivo knee position at each flexion angle. With this series of knee models, the location of the tibiofemoral cartilage contact, size of the contact area, cartilage thickness at the contact area, and magnitude of the cartilage contact deformation were compared between intact and ACLdeficient knees.Results. Rupture of the ACL changed the cartilage contact biomechanics between 0°and 60°of flexion in the medial compartment of the knee. Compared with the contralateral knee, the location of peak cartilage contact deformation on the tibial plateaus was more posterior and lateral, the contact area was smaller, the average cartilage thickness at the tibial cartilage contact area was thinner, and the resultant magnitude of cartilage contact deformation was increased. Similar changes were observed in the lateral compartment, with increased cartilage contact deformation from 0°to 30°of knee flexion in the presence of ACL deficiency.Conclusion. ACL deficiency alters the in vivo cartilage contact biomechanics by shifting the contact location to smaller regions of thinner cartilage and by increasing the magnitude of the cartilage contact deformation.