This study was conducted to evaluate the effect of applied load on the magnitude, direction, and point of tibial intersection of the in situ forces of the anteromedial (AM) and posterolateral (PL) bands of the human anterior cruciate ligament (ACL) at 30 degrees and 90 degrees of knee flexion. An Instron was used to apply a 100 N anterior shear force to 11 human cadaver knees, 6 at 30 degrees of knee flexion and 5 at 90 degrees of knee flexion. A Universal Force Sensor (UFS) recorded the resultant 6 degree-of freedom (DOF) forces/moments. Each specimen then underwent serial removal of the AM and PL bands. With the knee limited to 1 DOF (anteroposterior), tests were performed before and after each structure was removed. Because the path was identical in each test, the principle of superposition was applied. Thus, the difference between the resultant forces could be attributed to the force carried by the structure just removed. The magnitudes of force in the ACL at 30 degrees and 90 degrees of knee flexion were 114.1 +/- 7.4 N and 90.8 +/- 8.3 N, respectively (P < 0.05). At 30 degrees, the AM and PL bundles carried 95% and 4% of the total ACL force, respectively. At 90 degrees, the AM and PL bands carried 85% and 13%, respectively (P < 0.05). The direction of the in situ force in the whole ACL as well as its two bands correlated with the anatomic orientation of the ligament.(ABSTRACT TRUNCATED AT 250 WORDS)