Glyco-functionalization of ECM mimics, influence in morphology and cell behaviour Authors

Abstract

Current tissue engineering strategies primarily focus on replicating mechanical properties of the extracellular matrix (ECM); however, several studies have underscored the critical role of the ECM biochemical cues in developing functional tissue substitutes. Among these, glycans are known to play a key role in regulating cell fate. In this study we developed ECM mimetics glyco-conjugated with two different glycans, -D-glucopyranose and -D-galactopyranose, to investigate their influence on morphology and cell behaviour. The ECM mimics were generated crosslinking glycosylated gelatin samples, functionalised with different glycans by reductive amination, and hyaluronic acid. The crosslinking was performed by previous functionalization of gelatin and hyaluronic acid with tyramine, to enable enzymatic crosslinking via horseradish peroxidase (HRP) and hydrogen peroxide (H₂O₂). The glyco-conjugated hydrogels exhibited markedly different morphologies, characterized by increased fibrous content, smaller pore sizes, and more wrinkled surfaces. Bone marrow-derived mesenchymal stem cells (BM-MSCs) seeded in hydrogels functionalized -D-glucose and -Dgalactose exhibited a more elongated morphology.