Enzymatically Crosslinked Hybrid Hydrogels from Ultrasound-Decellularized Spinal Cord Meninges and Hydrophilic Silk...

Hydrogels combining the biochemical complexity of the native extracellular matrix (ECM) with the tunable properties of protein-based biomaterials are promising for neural tissue engineering. In this study, decellularized spinal cord meninges (dSCM) were combined with water-soluble hydrophilic silk fibroin (hSF) and enzymatically crosslinked using a horseradish peroxidase/H2O2 system to develop composite hydrogels. A detergent-free, sonication-assisted decellularization method effectively removed cellular components while preserving matrix integrity, reducing residual double-stranded DNA to below 50 ng mg-1 dry weight and retaining key ECM constituents, including collagen and glycosaminoglycans. Hydrogels prepared at different dSCM:hSF ratios showed composition-dependent structural and mechanical behavior, with the 1:0.5 and 1:1 formulations exhibiting the most favorable compressive stiffness and viscoelastic performance. Structural, thermal, and morphological analyses further indicated that hSF incorporation improved matrix stability and contributed to more controlled swelling and degradation behavior. Biological evaluation showed that the 1:0.5 formulation promoted neovascularization in the chorioallantoic membrane assay without evident adverse inflammatory response. In addition, SH-SY5Y cells maintained high viability and showed increased expression of the neuronal-associated markers β-III tubulin and MAP2 over time. Overall, these findings suggest that dSCM:hSF hydrogels provide a promising platform for neural tissue engineering.