Decellularized bovine intervertebral disc as a natural scaffold for xenogenic cell studies

Low back pain that is associated with disc degeneration contributes to a huge economic burden in the worldwide healthcare system. Traditional methods, such as spinal fusion, have been adopted to relieve mechanical back pain, but this is compromised by decreased spinal motion. Tissue engineering has attracted much attention, and aims to correct the changes fundamentally occurring in the discs by a combination of cell biology, molecular biology and engineering. Synthetic materials including poly(l-lactic acid) or poly(glycolic acid) and biomolecules like hyaluronic acid or collagen have been adopted in the development of disc scaffolds for studying therapeutic approaches. Nevertheless, the complex biological and mechanical environment of the intervertebral disc (IVD) makes the synthesis of an artificial IVD with biomaterials a difficult task. Thus the aim of this study was to develop a natural disc scaffold for culturing disc cells for future development of biological disc constructs. We adopted a combination of currently used decellularization techniques to decellularize bovine IVD to create a complete endplate-to-endplate IVD scaffold. By altering the chemical and physical decellularization parameters, we reported the removal of up to 70% of the endogenous cells, and were able to preserve the glycosaminoglycan content, collagen fibril architecture and mechanical properties of the discs. The reintroduction of nucleus pulposus cells into the scaffold indicated a high survival rate over 7 days, with cell penetration. We have shown here that conventional methods used for decellularizing thin tissues can also be applied to large organs, such as IVD. Our findings suggest the potential of using decellularized IVD as a scaffold for IVD bioengineering and culturing of cells in the context of the IVD niche.