In order to evaluate the contribution of the knee ligaments to restrain joint motions, knowledge about their structural properties is required. Due to the variable relative insertion orientation of the ligaments during knee motion, however, different fiber bundles are recruited, each with their specific mechanical properties. Hence, the structural properties vary as a function of knee motion. For this reason, a relationship between the structural tensile properties and the relative insertion orientation is required in order to define the role of the ligaments in knee mechanics. In the present study, this relationship is determined by performing a series of tensile tests in which the relative orientations of the insertion sites of human knee bone-ligament-bone preparations were varied systematically. The experimentally obtained stiffness was significantly affected by the relative orientation of the insertion sites, but more profoundly for the anterior and posterior cruciate ligaments (ACL and PCL) as compared to the medial and lateral collateral ligaments (MCL and LCL). The average decreases in stiffness per 5 degrees tilt of the insertion sites were estimated at -11.6 +/- 3.5 N mm-1 (ACL), -20.9 +/- 2.7 N mm-1 (PCL), -2.6 +/- 0.9 N mm-1 (MCL) and -3.7 +/- 0.3 N mm-1 (LCL). For the PCL and the MCL these changes in stiffness with tilt were rather insensitive to the side of the femoral insertion site which was lifted. The ACL and the LCL, conversely, displayed significant differences in stiffness changes between the different tilt directions.(ABSTRACT TRUNCATED AT 250 WORDS)