The spatial and bulk water equilibrium partition and fluid content were determined for normal adult bovine articular cartilage as a function of pH, temperature, and geometric confinement. Water partition averaged 60 ± 7% at neutral pH and 37°C and increased with decreasing pH and increasing temperature without a concomitant change in fluid content. The variation in water partition appeared to be a result of local conformation changes in the collagen fibril ultrastructure causing a transfer between free and trapped water volume. Removal of the lateral and subchondral bone geometric constraints caused an increase in both the water partition and fluid content. However, this partiton variation could be accounted for solely from a change in free fluid volume. These results suggest that in articular cartilage the proteoglycan-collagen interaction may be an important mechanism for controlling the partition of water between a freely exchangeable space and a space allowing no fluid exchange.