Injectable decellularized nucleus pulposus-based cell delivery system for differentiation of adipose-derived stem cells and nucleus pulposus regeneration

Abstract

Stem cell-based tissue engineering is a promising treatment for intervertebral disc (IVD) degeneration. A bio-scaffold that can maintain the function of transplanted cells and possesses favorable mechanical properties is needed in tissue engineering. Decellularized nucleus pulposus (dNP) has the potential to be a suitable bio-scaffold because it mimics the native nucleus pulposus (NP) composition. However, matrix loss during decellularization and difficulty in transplantation limit the clinical application of dNP scaffolds. In this study, we fabricated an injectable dNP-based cell delivery system (NPCS) and evaluated its properties by assessing the microstructure, biochemical composition, water content, biosafety, biostability, and mechanical properties. We also investigated the stimulatory effects of the bio-scaffold on the NP-like differentiation of adipose-derived stem cells (ADSCs) in vitro and the regenerative effects of the NPCS on degenerated NP in an in vivo animal model. The results showed that approximately 68% and 43% of the collagen and sGAG, respectively, remained in the NPCS after 30 days. The NPCS also showed mechanical properties similar to those of fresh NP. In addition, the NPCS was biocompatible and able to induce NP-like differentiation and extracellular matrix (ECM) synthesis in ADSCs. The disc height index (almost 81%) and the MRI index (349.05 ± 38.48) of the NPCS-treated NP were significantly higher than those of the degenerated NP after 16 weeks. The NPCS also partly restored the ECM content and the structure of degenerated NP in vivo. Our NPCS has good biological and mechanical properties and has the ability to promote the regeneration of degenerated NP. Statement of SignificanceNucleus pulposus (NP) degeneration is usually the origin of intervertebral disc degeneration. Stem cell-based tissue engineering is a promising treatment for NP regeneration. Bio-scaffolds which have favorable biological and mechanical properties are needed in tissue engineering. Decellularized NP (dNP) scaffold is a potential choice for tissue engineering, but the difficulty in balancing complete decellularization and retaining ECM limits its usage. Instead of choosing different decellularization protocols, we complementing the sGAG lost during decellularization by cross-linking via genipin and fabricating an injectable dNP-based cell delivery system (NPCS) which has similar components as the native NP. We also investigated the biological and mechanical properties of the NPCS in vitro and verified its regenerative effects on degenerated IVDs in an animal model.