Investigation of nano-mechanical properties of annulus fibrosus using atomic force microscopy

We describe the use of atomic force microscopy (AFM) to investigate the nanomechanical properties of annulus fibrosus (AF)-the outer fibrous layer of an intervertebral disc (IVD) encapsulating the inner jelly-like mass known as the nucleus pulposus (NP). Disk disease, degenerated discs, slipped discs, and herniated discs are common terms often linked to back pain and are caused due to degeneration of IVD. Due to the variations in the structure and biochemical composition of the IVD, studies of macromechanical properties in the motion segment or AF may lack all significant nanomechanical responses or behaviors. Existing studies do not report the micro or nano level of mechanics of IVD components and whether the nanomechanics of this tissue mimic its macromechanical behavior is not known. Our studies used AFM to investigate the regional micromechanical properties of the AF that have been otherwise difficult due to small sample size of the tissue. Five different zones including peripheral and central were tested mechanically as well as biochemically. Qualitative biochemical staining and quantitative values of nanomechanical properties of different zones are compared and discussed in detail. The results of nanomechanical investigations described in this study not only reveal its mimic at macroscopic level, they represent an important step towards establishing a framework for testing and comparing tissue engineered IVD replacements with native tissues.