A dural strategy for 3D-printed SilMA hydrogels modification: Nanofiber reinforcement and PEO-induced porosity

Hydrogels have emerged as promising scaffolds for cartilage tissue engineering due to their structural mimicry of native articular cartilage extracellular matrix. However, conventional hydrogels typically exhibit only nanoscale porosity and poor mechanical properties, which limit nutrient delivery, metabolic waste exchange, and structural fidelity. To address these challenges, we developed an innovative cell-laden porous methacrylated silk fibroin (SilMA) hydrogel system with biomechanical reinforcement through 3D bioprinting strategy. The porous architecture was created through a water-in-water emulsification strategy employing poly(ethylene oxide) (PEO) as a sacrificial template. This pore-forming process resulted in remarkable structural modulation, achieving over 100% increase in average pore diameter and 75% enhancement in overall porosity compared to hydrogels without PEO application. However, this structural modification compromised the compressive modulus by approximately 50%. Therefore, homogenized electrospun silk fibroin nanofibers (NFs) were incorporated into the bioink to implement the mechanical properties and optimize surface topography. The introduction of NFs (1-2 wt%) not only recovered the compressive strength and modulus (near to SilMA hydrogels) but also improved the 3D printability PEO/SilMA hydrogels. Additionally, the hydrogel demonstrated excellent biocompatibility and markedly upregulated chondrogenic-related gene expression, including Collagen II, Aggrecan, and Sox9. Furthermore, the subcutaneous implantation experiments in NOD/SCID rats further confirmed the potential of PEO/NFs/SilMA hydrogels in promoting cartilage formation. Therefore, this study proposes a promising dual-strategy approach for cartilage tissue engineering, integrating NFs reinforcement and PEO-induced porosity.