Harnessing Thrombospondin-1-Enabled Decellularized Nucleus Pulposus Matrices and Elastin-Like Recombinamers to Rebuild an Avascular Analogue of the Intervertebral Disc.

Abstract

With the degeneration of the intervertebral disc (IVD), the ingrowth of vascular and neural structures occurs. Both nerves and blood vessels engage in the development of inflammation and the onset of discogenic pain. The present study aimed to produce a hierarchical biomaterial capable of inhibiting angiogenesis by emulating the microenvironment of non-degenerated IVDs. To this end, we have incorporated an angiogenesis modulator-thrombospondin-1 (TSP-1) into a three-dimensional (3D) hydrogel network containing decellularized nucleus pulposus (dNPs) and azide-cyclooctyne modified elastin-like recombinamers (ELRs). Following the decellularization of nucleus pulposus (NPs) isolated from bovine tissues, pre-gels (pGs) were assembled based on the acid-pepsin extraction of soluble collagens found in the dNPs. Given the inherent affinity of these macromolecules to TSP-1, which was corroborated by immunohistochemical analysis and FT-IR spectroscopy, the pGs were supplemented with two concentrations of TSP-1. Angiogenesis was evaluated using the chick chorioallantoic membrane (CAM) in vivo model. Conjugation of TSP-1 with the pGs resulted in a synergistic suppression of blood vessel formation. Complexation with the ELRs improved the viscoelastic moduli and the structural stability of the hydrogels, which maintained their hydration and osmolarity properties due to the presence of the dNPs. When placed in direct contact with human primary fibroblasts, the materials displayed high cytocompatibility and tunable degradation rates. Our findings indicate that TSP-1-enabled dNP-derived pGs inhibit angiogenesis in vivo, while the presence of the ELRs aids in improving the mechanical properties of the hydrogels, thus providing a platform for rebuilding an avascular analogue of the healthy IVD.