Although tissue engineering has attracted increasing attention for the treatment of degenerative intervertebral disc disease, the biochemical properties, structural organization, and mechanical characteristics of annulus fibrosus tissue have restricted progress. Differentiation of annulus fibrosus-derived stem cells (AFSCs) can be regulated by the elasticity of substrates such as poly(ether carbonate urethane)urea (PECUU). Decellularized annulus fibrosus matrix (DAFM) has good biocompatibility and biodegradability, making it suitable for cell adhesion, proliferation, and differentiation. In this study, we used a coaxial electrospinning method to synthesize DAFM/PECUU-blended fibrous scaffolds with elasticities approximating that of native inner and outer annulus fibrosus tissue. AFSCs cultured on DAFM/PECUU-blended fibrous scaffolds exhibited increased collagen type I gene expression with increasing elasticity of the scaffold material; notably, collagen type II and aggrecan gene expression exhibited the opposite trend. Regarding extracellular matrix secretion, collagen type I content gradually increased with substrate elasticity, while collagen type II and aggrecan contents decreased. In vivo evaluations employing magnetic resonance imaging, hematoxylin and eosin staining, and immunohistochemistry indicated that DAFM/PECUU-blended fibrous scaffolds could effectively repair defects of annulus fibrosus tissue. Our findings provide a theoretical and practical basis for the development of bionic annulus fibrosus tissue that closely mimics the biological properties, mechanical function, and matrix composition of native tissue.