The purpose of this study was to investigate the effect of spinal instrumentation on the intradiscal pressure (IDP) within the fixed motion segment. In vitro biomechanical testing was performed in six single functional spinal units of fresh calf lumbar spines using a pressure needle transducer. Various loads were applied by a materials testing system device. In addition to intact spine (control), anterior spinal instrumentation (ASI) and pedicle screw fixation (PS) constructs, as well as destabilized spine were tested. Relative to the control, the destabilized spine tended to have an increased IDP; by 15% in axial compression and by 9-36% in flexion-extension. Compared to the control, PS decreased the IDP by 23% in axial loading and 51% in extension loading and increased it by 60% in flexion for each loading. ASI decreased the IDP by 32% in flexion and 1% in extension. Lateral bending produced symmetrical changes of IDP in the control and destabilized spine, but no change in the PS construct. The IDP of the ASI construct was decreased by 77% in ipsilateral bending and increased by 22% in contralateral bending. These results demonstrated that eccentric loading from the spinal instruments increased IDP and significant disc pressure may still exist despite an increase in motion segment stiffness after lumbar stabilization.