A non‐invasive methodology (based on video microscopy, optimized digital image correlation and thin plate spline smoothing technique) has been developed to determine the intrinsic tissue stiffness (H_a) and the intrinsic fixed charge density (c₀^F) distribution for hydrated soft tissues such as articular cartilage. Using this technique, the depth‐dependent inhomogeneous parameters H_a(z) and c₀^F(z) were determined for young bovine cartilage and incorporated into a triphasic mixture model. This model was then used to predict the mechanical and electrochemical events (stress, strain, fluid/osmotic pressure, and electrical potentials) inside the tissue specimen under a confined compression stress relaxation test. The integration of experimental measurements with theoretical analyses can help to understand the unique material behaviors of articular cartilage. Coupled with biological assays of cell‐scale biosynthesis, there is also a great potential in the future to study chondrocyte mechanotransduction in situ with a new level of specificity.