The objective of this study was to determine the equilibrium confined compression modulus of bovine articular cartilage as it varies with depth from the articular surface. Osteochondral samples were compressed by 8, 16, 24, and 32% of the cartilage thickness and allowed to equilibrate. Intratissue displacement within the cartilage was measured with use of fluorescently labeled chondrocyte nuclei as intrinsic, fiducial markers. Axial strain was then calculated in nine sequential 125 microns thick cartilage layers comprising the superficial 1,125 microns and in a 250 microns thick layer of cartilage adjacent to the cartilage-bone interface. Adjacent osteochondral cores were also tested in confined compression to determine the equilibrium stresses required to achieve the same levels of compression. Stress-strain data for each layer of each sample were fit to a finite deformation stress-strain relation to determine the equilibrium confined compression modulus in each tissue layer. The compressive modulus increased significantly with depth from the articular surface and ranged from 0.079 +/- 0.039 MPa in the superficial layer to 1.14 +/- 0.44 MPa in the ninth layer. The deepest layer 250 microns thick, had a modulus of 2.10 +/- 2.69 MPa. These moduli were markedly different from the apparent "homogeneous" modulus for full-thickness cartilage (0.38 +/- 0.12 MPa) and ranged from 21 to 560% of that value. The relatively low moduli and the compression-induced stiffening of the superficial layers suggest that these layers greatly affect the biomechanical behavior of cartilage, such as during confined compression testing. The delineation of the depth-dependent modulus provides a basis for detailed study of the relationship between the composition, structure, and function of cartilage in such processes as aging, repair, and degeneration.