The relation between mechanical loading of cartilage and chondrocyte activity in vivo may be mediated by several physical transduction mechanisms including:​ cell deformation, hydrostatic pressure gradients, fluid flow, streaming currents, and physicochemical changes. We have developed an organ culture system designed to study chondrocyte biosynthetic response to such physical stimuli. This study focuses on the effects of static compression and physicochemical changes. Cartilage disks harvested from the reserve zone of the epiphyseal plate of 1–2-week-old calves were subjected to static compressive stresses of 0–3 MPa in unconfined compression and the incorporation of [³⁵S]sulfate and [³H]proline was measured during the 12-h loading period. Incorporation of both proline and sulfate decreased monotonically with increasing stress. Compressive loading also produces physicochemical changes including a decreased water content and increased matrix fixed-charge density, with a concomitant increase in interstitial counterion concentrations (e.g., K⁺, H⁺) and decreased coion concentrations (e.g., SO₄²⁻). We therefore examined the possibility that specific changes in interstitial mobile ion concentrations may be linked to chondrocyte response to static compression by measuring biosynthesis in the absence of mechanical compression while independently altering the SO₄²⁻, K⁺, and H⁺ composition of the bathing medium. We found that proline and sulfate incorporation were strongly dependent on pH, but independent of [SO₄²⁻]and [K⁺] in the range studied. These results suggest that compression-induced changes in local, interstitial pH may account for the observed biosynthetic response to static compression.