The objective of this study was to determine the biomechanical effect of graft tensioning during reconstruction of the anterior cruciate ligament. We evaluated the magnitude of the tensioning force (22 or 67 N), the flexion angle at which the tension was applied (extension or 30° of flexion), and the direction of application of the tensioning force (proximal, distal, or distal with a posterior force simultaneously applied to the tibia) on 10 fresh cadaveric knees. The anterior cruciate ligament was reconstructed using a bone-patellar tendon-bone graft. The graft was then temporarily fixed during the application of each of 12 combinations of tensioning variables listed above. After each fixation, graft force and joint motion were measured during anterior tibial loads. Tensioning direction and the flexion angle significantly affected graft force and joint motion, while the magnitude of the graft tensioning did not. Graft forces were greater when the tensioning was applied at 30° of flexion. Compared with distal tensioning with and without posterior tibial force, graft forces with proximal tensioning were greater in extension and lower in flexion. The position of the tibia relative to the femur was posterior and externally rotated, compared with normal, for all combinations of tensioning variables in both unloaded and anterior load states.