The stiffness of motion segments, together with muscle actions, stabilizes the spinal column. The objective of this study was to compare the experimentally measured load-displacement behavior of porcine lumbar motion segments in vitro with physiological axial compressive preloads of 0, 200 and 400 N equilibrated in a physiological fluid environment, for small displacements about the neutral posture. These preloads are hypothesized to increase stiffness, hysteresis and linearity of the load-displacement behavior. At each preload, displacements in each of six degrees of freedom (+/-0.3 mm AP and lateral translations, +/-0.2 mm axial translation, +/-1 degrees lateral bending and +/-0.8 degrees flexion/extension and torsional rotations) were imposed. The resulting forces and moments were recorded. Tests were repeated after removal of posterior elements. Using least squares, the forces at the vertebral body center were related to the displacements by a symmetric 6 x 6 stiffness matrix. Six diagonal and two off-diagonal load-displacement relationships were examined for differences in stiffness, linearity and hysteresis in each testing condition. Mean values of the diagonal terms of the stiffness matrix for intact porcine motion segments increased significantly by an average factor of 2.2 and 2.9 with 200 and 400 N axial compression respectively (p<0.001). Increases for isolated disc specimens averaged 4.6 and 6.9 times with 200 and 400 N preload (p<0.001). Changes in hysteresis correlated with the changes in stiffness. The load-displacement relationships were progressively more linear with increasing preload (R(2)=0.82, 0.97 and 0.98 at 0, 200 and 400 N axial compression respectively). Motion segment and disc load-displacement behaviors were stiffer, more linear and had greater hysteresis with axial compressive preloads.