Depth-dependent anisotropy of the micromechanical properties of the extracellular and pericellular matrices of articular cartilage evaluated via atomic force microscopy.

The extracellular matrix (ECM) of articular cartilage is structurally and mechanically inhomogeneous and anisotropic, exhibiting variations in composition, collagen fiber architecture, and pericellular matrix (PCM) morphology among the different zones (superficial, middle, and deep). Joint loading exposes chondrocytes to a complex biomechanical environment, as the microscale mechanical environment of the chondrocyte depends on the relative properties of its PCM and local ECM. ECM anisotropy and chondrocyte deformation are influenced by the split-line direction, the preferred collagen fiber orientation parallel to the articular surface. While previous studies have demonstrated that cartilage macroscale properties vary with depth and the direction of loading relative to the split-line direction, the potential anisotropic behavior of the ECM and PCM at the microscale has yet to be examined. The goal of this study was to characterize the depth and directional dependence of the microscale biomechanical properties of porcine cartilage ECM and PCM in situ. Cartilage was cryosectioned to generate samples oriented parallel and perpendicular to the split-line direction and normal to the articular surface. Atomic force microscopy (AFM)-based stiffness mapping was utilized to measure ECM and PCM microscale elastic properties in all three directions within each zone. Distinct anisotropy in ECM elastic moduli was observed in the superficial and deep zones, while the middle zone exhibited subtle anisotropy. PCM elastic moduli exhibited zonal uniformity with depth and directional dependence when pooled across the zones. These findings provide new evidence for mechanical inhomogeneity and anisotropy at the microscale in articular cartilage.