Abstract
Silk fibroin (SF) was enzymatically crosslinked with tyramine-substituted silk fibroin (SF-TA) or gelatin (G-TA) to fabricate hybrid hydrogels with tunable gelation kinetics, mechanical properties and bioactivity. Horseradish peroxidase (HRP)/hydrogen peroxide (H2O2) mediated crosslinking of SF in physiological buffers results in slow gelation and limited mechanical properties. Moreover, SF lacks cell attachment sequences, leading to poor cell-material interactions. These shortcomings can limit the uses of enzymatically crosslinked silk hydrogels in injectable tissue fillings, 3D bioprinting or cell microencapsulation, where rapid gelation and high bioactivity are desired. Here SF/SF-TA and SF/G-TA composite hydrogels were characterized for hydrogel properties and the influence of conjugated cyclic arginine-glycine-aspartic acid (RGD) peptide or G-TA content on bioactivity was explored. Both SF-TA and G-TA significantly increased gelation kinetics, improved mechanical properties and delayed enzymatic degradation in a concentration-dependent manner. β-Sheet formation and hydrogel stiffening were accelerated by SF-TA content but delayed by G-TA. Both cyclic RGD and G-TA significantly improved morphology and metabolic activity of human mesenchymal stem cells (hMSCs) cultured on or encapsulated in composite hydrogels. The hydrogel formulations introduced in this study provide improved control of gel formation and properties, along with biocompatible systems that can be utilized in tissue engineering and cell delivery applications.
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