The objective of this study was to assess mechano-biological response of articular cartilage when subjected to a single impact stress. Mature bovine cartilage explants were impacted with peak stresses ranging from 10 to 60 MPa at a stress rate of 350 MPa/s. Water loss, matrix axial deformation, dynamic impact modulus (DIM), and cell viability were measured immediately after impaction. The water loss through the articular surface (AS) was small and ranged from 1% to 6% with increasing peak stress. The corresponding axial strains ranged from 2.5% to 25%, respectively, while the DIM was 455.9±111.9 MPa. Chondrocyte death started at the articular surface and increased in depth to a maximum of 6% (70 μm) of the cartilage thickness at the highest stress. We found that the volumetric (axial) strain was more than twice the amount of water loss at the highest peak stress. Furthermore, specimens impacted such that the interstitial water was forced through the deep zone (DZ) had less water loss, a higher DIM, and no cell death. These findings appear to be due to matrix compaction in the superficial region causing higher compressive strains to occur at the surface rather than in the deeper zones.