Strain in the anteromedial fibers of the anterior cruciate ligament [ACL(am)] was studied in six cadaver knees. ACL(am) strain was measured in five knees during the application of isometric quadriceps forces alone and simultaneously applied isometric quadriceps and hamstrings forces at 10 degrees increments from 0 degrees to 90 degrees of knee flexion. ACL(am) strain during muscle loading was measured with respect to the ACL(am) strain measured with the knee in its resting position (neutral or near neutral position). A sixth knee was used to investigate the reproducibility of the resting position and quadriceps-induced ACL(am) strains. The strains induced in the ACL(am) by the quadriceps were significantly greater than 0 at knee flexion angles from 0 to 40 degrees and not significantly different from 0 for 50 to 90 degrees. The ACL(am) strains induced by simultaneously applied hamstrings and quadriceps forces were not significantly different from 0 at any of the knee flexion angles tested. Simultaneously applied hamstrings and quadriceps forces significantly reduced ACL(am) strain at 10, 20, and 90 degrees of knee flexion compared to the ACL(am) strain induced by quadriceps forces alone. The hamstrings are potentially capable of both significantly reducing and negating quadriceps-induced ACL(am) strain at 10 and 20 degrees of knee flexion.