Altered patellofemoral joint contact pressures are thought to contribute to patellofemoral joint symptoms. However, little is known about the relationship between tibiofemoral joint kinematics and patellofemoral joint contact pressures. The objective of this paper was to investigate the effect of tibiofemoral joint kinematics on patellofemoral joint pressures using an established in vitro robotic testing experimental setup. Eight cadaveric knee specimens were tested at 0 degrees, 30 degrees, 60 degrees, 90 degrees, and 120 degrees of flexion under an isolated quadriceps load of 400 N and a combined quadriceps/hamstrings load of 400 N/200 N. Tibiofemoral joint kinematics were measured by the robot and contact pressures by a TekScan pressure sensor. The isolated quadriceps loading caused anterior translation and internal rotation of the tibia up to 60 degrees of flexion and posterior translation and external rotation of the tibia beyond 60 degrees. The co-contraction of the hamstring muscles caused a posterior translation and external rotation of the tibia relative to the motion of the tibia under the quadriceps load. Correspondingly, the contact pressures were elevated significantly at all flexion angles. For example, at 60 degrees of flexion, the hamstrings co-contraction increased the posterior tibial translation by approximately 2.8 mm and external tibial rotation by approximately 3.6 degrees. The peak contact pressure increased from 1.4+/-0.8 to 1.7+/-1.0 MPa, a 15% increase. The elevated contact pressures after hamstrings co-contraction indicates an intrinsic relation between the tibiofemoral joint kinematics and the patellofemoral joint biomechanics. An increase in posterior tibial translation and external rotation is accompanied by an increase in contact pressure in the patellofemoral joint. These results imply that excessive strength conditioning with the hamstring muscles might not be beneficial to the patellofemoral joint. Knee pathology that causes an increase in tibial posterior translation and external rotation might contribute to degeneration of the patellofemoral joint. These results suggest that conservative treatment of posterior cruciate ligament injury should be reconsidered.