Hyaluronan composite bioink preserves nucleus pulposus cell phenotype in a stiffness-dependent manner

Intervertebral disc degeneration is a major cause of neck and back pain, representing a significant global socioeconomic burden. The polysaccharide hyaluronan is key to maintaining disc physiology and mediating disc disease through its structural and biological roles in the nucleus pulposus, a component of the intervertebral disc highly susceptible to degeneration. In this study, we introduce a novel composite bioink designed for extrusion bioprinting of structures resembling the nucleus pulposus. Our bioink combines levels of hyaluronic acid and collagen that approach physiological concentrations and effectively mimics the disc's hydrated and mechanically resilient environment. We modulated the composite's mechanical properties through the tyramination of hyaluronic acid and subsequent photocrosslinking, influencing morphology and gene expression of embedded bovine nucleus pulposus cells. This allows us to replicate a range of properties from healthy to degenerated human nucleus pulposus, which would be challenging to achieve with traditional cell culture and in vivo models. Our results show that modulating hyaluronan physico-chemical properties influenced embedded cell phenotype. The outcomes of this study inform the future design of biomaterials for the modelling of disc disease and regeneration, and present a versatile platform that can be readily integrated with other biofabricated components to form engineered intervertebral disc-like structures.