The effects of mechanical stimuli on TMJ disc cells have yet to be investigated. This study examined for the first time the effect of constant and intermittent hydrostatic pressure (HP) on TMJ disc cells. Guided by studies on articular chondrocytes, the chosen amplitude was 10 MPa, the frequency was 1 Hz for intermittent HP, and the duration was 4 h. A one-time application of the HP stimulus was applied in 2-D and 3-D for gene expression studies. A duty cycle of 2 days on, 1 day off for 1 week of HP stimulus was used for biochemical content studies. In monolayer, the intermittent HP regimen increased collagen II expression, while constant HP increased collagen I expression when compared to the non-loaded control. However, the overall expression of collagen I was much higher than collagen II in both constant and intermittent HP. The expression results correlated well with gross morphology, histology, and biochemical content. At Week 1, the intermittent HP group had a lower content of collagen, 7.5 +/- 0.2 microg/construct, than the non-loaded control group, 18.2 +/- 4.0 microg/construct. The constant HP group showed the highest amount of collagen, 24.5 +/- 1.6 microg/construct. These data show that constant HP at 10 MPa for 4 h produces more collagen I than do the non-loaded control or intermittent HP at 10 MPa and 1 Hz.