Nanomechanical mapping of the osteochondral interface with contact resonance force microscopy and nanoindentation

The bone–cartilage, or osteochondral, interface resists remarkably high shear stresses and rarely fails, yet its mechanical characteristics are largely unknown. A complete understanding of this hierarchical system requires mechanical-property information at the length scales of both the interface and the connecting tissues. Here, we combined nanoindentation and atomic force microscopy (AFM) methods to investigate the multiscale mechanical properties across the osteochondral region. The nanoindentation modulus M ranged from that of the subchondral bone (M = 22.8 ± 1.8 GPa) to that of hyaline articular cartilage embedded in PMMA (M = 5.7 ± 1.0 GPa) across a narrow transition region <5 μm wide. Contact resonance force microscopy (CR-FM), which measures the frequency and quality factor of the AFM cantilever’s vibrational resonance in contact mode, was used to determine the relative storage modulus and loss tangent of the osteochondral interface. With better spatial resolution than nanoindentation, CR-FM measurements indicated an even narrower interface width of 2.3 ± 1.2 μm. Furthermore, CR-FM revealed a 24% increase in the viscoelastic loss tangent from the articular calcified cartilage into the PMMA-embedded hyaline articular cartilage. Quantitative backscattered electron imaging provided complementary measurement of mineral content. Our results provide insight into the multiscale functionality of the osteochondral interface that will advance understanding of disease states such as osteoarthritis and aid in the development of biomimetic interfaces.