Dual Modification of Collagen Hydrogels Using UVA-Riboflavin Crosslinking and Silk Fibroin Incorporation to Enhance Mechanical Properties, Stability, and Printability.
Vijayan Nithyashri Muthu NM, Babu Diya Pillai DP, Caner Beste B, Shyam Rohin R et al.
Type I collagen is a biomaterial of preferred choice due to its biocompatibility, biodegradability, and ability to support cell adhesion and proliferation. However, the use of collagen hydrogels in tissue engineering (TE) is limited due to poor mechanical properties and low stability. This study introduces a novel dual-modification strategy to enhance collagen properties by combining UVA-mediated riboflavin/sodium persulfate (RF/SPS) photocrosslinking with the incorporation of unsonicated silk fibroin (SF), an approach not previously reported. Collagen hydrogels were fabricated in four variations: uncrosslinked collagen (UXL-C), UVA-mediated RF/SPS photocrosslinked collagen (XL-C), SF incorporated UVA-RF/SPS photocrosslinked collagen using two different collagen:SF ratios (XL-CS 1:1 w/w) and (XL-CS 1:2 w/w). Mechanical testing and stability results showed that combining UVA-RF/SPS photocrosslinking and SF incorporation significantly enhanced (p < 0.05) the compressive modulus and increased mass-retention of collagen hydrogels suggesting that the addition of unsonicated SF improves the mechanical properties and stability of photochemically crosslinked collagen hydrogels. All hydrogel compositions exhibit shear-thinning behavior demonstrating applicability for extrusion-based 3D printing via Freeform Reversible Embedding of Suspended Hydrogels (FRESH)-based methodology. 3D printed SF-incorporated collagen constructs showed improved shape fidelity compared to constructs printed with collagen-only inks. In vitro cell culture studies revealed that all hydrogel formulations supported cell viability and metabolic activity over 7 days of culture. Together, these findings demonstrate that combining UVA-mediated RF/SPS photocrosslinking and unsonicated SF incorporation is a viable strategy to improve the mechanical properties, stability, and printability of collagen hydrogels for biomedical and TE applications.