Brief exposure to inflammation may have significant consequences on cell biomechanical properties, regardless of the recovery microenvironment to non-inflammatory levels, in vitro. In this study, we chose to measure the gene expression of inflammatory cytokine IL-1b as a mediator of the inflammatory stimulation experiment. IL-1b has been shown to be upregulated in NP cells during disc degeneration in vivo and upregulated in response to LPS or TNF-a treatment in vitro. The influence of inflammation on the osmotic behavior of NP cells at a wide osmotic range mimicking the in situ osmolarity of the intervertebral disc during physiological activity was examined, where osmotic loads in the range of 450–550 mOsm/L or 100–200 mOsm/L have been reported in normal vs. degenerated discs, respectively. Step osmotic loading resulted in transient cell-volume responses followed by sustained equilibrium. The steady-state volume response of cells in each treatment group demonstrated a linear behavior over the applied osmolarities consistent with the Boyle-van’t Hoff relation for a perfect osmometer which is expected when regulatory volume changes are negligible. Loading with solutions at physiologic temperature and composition could help elucidate the effects of inflammation on active cellular response in future studies. Untreated NP cells exhibited an average hydraulic permeability similar to that previously reported for articular chondrocytes, but the intracellular water content was lower than previously reported in chondrocytes . Cellular expression of Aqp-1, the water channel isoform reported to be most abundant in the human IVD, was found to decrease with inflammatory stimulation in NP cells. Consistent with our findings, Aqp-1 has been shown to be downregulated in murine epithelial cells in response to inflammatory treatment by LPS. Overall, increases in Aqp-1 expression are associated with increases in membrane water channels, which can accelerate the cell swelling rate by providing a greater number of conduits for water transport. Aqp-1 expression at the cellular level was significantly lower in LPS and TNF-a treated and recovery groups compared to untreated controls. These findings suggest that the observed permeability differences in inflammatory-stimulated cells are not directly correlated with Aqp-1 cellular expression in NP cells. Alternatively, other aquaporin isoforms, not investigated in the current study, may be contributing to the permeability increases. Interestingly, we did not observe significant differences in Apq-1 gene expression between untreated and LPS treated cells at both time points, suggesting that inflammatory stimulation regulates Aqp-1 transcription more than gene expression in NP cells. We analyzed and quantified Aqp-1 expression using IHC on a large number of cells to statistically confirm the decreases in Aqp-1 expression due to inflammation.