Decellularized CNS-Derived ECM Hydrogels as Dual-Action Scaffolds for Angiogenesis and Neurogenesis

Abstract Neurodegenerative disorders of the central nervous system (CNS) remain a significant clinical challenge due to the limited regenerative capacity of neural tissues. Herein, bioactive scaffolds were developed from decellularized extracellular matrices (dECMs) derived from four CNS tissues: brain, spinal cord, spinal meninges, and optic nerve. The scaffolds were comprehensively characterized with respect to biochemical composition, mechanical properties, and biological performance. Optimized decellularization protocols efficiently removed cellular components while preserving key matrix constituents, including collagens and glycosaminoglycans (GAGs). The resulting dECM scaffolds exhibited distinct tissue-specific mechanical profiles; meninges (dM) and optic nerve (dO) scaffolds showed enhanced elasticity (>64% strain), whereas brain-derived (dB) scaffolds displayed higher stiffness in the lyophilized state. All scaffolds were highly porous (>60%) and hydrophilic, with swelling ratios exceeding 1000%. 3D culture studies using bEnd.3 endothelial and SH-SY5Y neuronal cells demonstrated favorable cytocompatibility (>65% viability) and tissue-dependent cellular responses. dM and dO scaffolds promoted slightly improved endothelial cell adhesion relative to dB and spinal cord (dC), while neuronal cell proliferation was more pronounced in dB, dC, and dO scaffolds. In addition, in ovo chick chorioallantoic membrane (CAM) assays confirmed the pro-angiogenic activity of all scaffold types. Collectively, these findings demonstrate that CNS-derived dECM scaffolds can support both angiogenic and neurogenic processes. This work provides the first comparative assessment of multiple CNS-derived dECMs in terms of their dual biofunctional potential and underscores the importance of tissue-specific ECM composition in guiding cell behavior for neurovascular tissue engineering applications.