The periodontal ligament (PDL) functions both in tension and in compression. The presence of an extensive vascular network inside the tissue suggests a significant contribution of the fluid phase to the mechanical response. This study examined the load response of bovine PDL under different pore pressure levels. A custom-made pressure chamber was constructed. Rod-shaped specimens comprising portions of dentine, bone, and intervening layer of PDL were extracted from bovine mandibular molars. The dentine ends of the specimens were secured to the actuator while the bone ends were affixed to the load cell. The entire assemblage was surrounded by the pressure chamber, which was then filled with saline. Specimens loaded at 1.0 Hz sinusoidal displacement were subjected to four different environmental fluid pressures (i.e., pressures of 0.0–1.0 MPa). The video images recorded during the tests were analyzed to determine whether or not fluid exchange between the PDL and the surrounding medium took place during mechanical loading. A value for the tissue’s apparent Poisson ratio was also determined. The following observations were made: (1) fluid was squeezed out and pumped into the ligament during the compressive and tensile loading phases, (2) the PDL was highly compressible, and (3) the pore pressure had no influence on the mechanical response of the PDL. The present tests emphasized the biphasic structure of PDL tissue, which should be considered as a porous solid matrix through which fluid can freely flow.