Mechanical properties of bovine articular cartilage under microscale indentation loading from atomic force microscopy

Atomic force microscopy (AFM) techniques have been increasingly used for investigating the mechanical properties of articular cartilage. According to the previous studies reporting the microscale Young's modulus under AFM indentation tests, the Hertz contact model has been employed with a sharp conical tip indenter. However, the non-linear microscale behaviour of articular cartilage could not be resolved by the standardized Hertz analysis using small and sharp atomic force microscope tips. Therefore, the objective of this study was to evaluate the microscale Young's modulus of articular cartilage more accurately through a non-Hertzian approach with a spherical tip of 5 microm diameter, and to characterize its microscale mechanical behaviour. This methodology adopted in the present study was proved by the consistent values between the microscale (2 per cent, about 9.3 kPa; 3 per cent, about 17.5kPa) and macroscale (2 per cent, about 8.3kPa; 3 per cent, about 18.3kPa) Young's moduli for 2 per cent and 3 per cent agarose gel (n = 100). Therefore, the microscale Young's modulus evaluated in this study is representative of more accurate measurements of cartilage stiffness at the 600 nm deformation level and corresponds to approximately 30.9 kPa (n = 100). Furthermore, on this level of the microscale deformation, articular cartilage showed depth-dependent and frequency-independent behaviour under AFM indentation loading. These findings reveal the microscale mechanical behaviour of articular cartilage more accurately and can be employed further to design microscale structures of chondrocyte-seeded scaffolds and tissue-engineered cartilage by evaluating their microscale properties.