Bioorthogonal Modification of Cell Derived Matrices by Metabolic Glycoengineering

Cell-derived matrices (CDMs) emerged as an attractive biomaterial in regenerative medicine. Here we present a strategy for site-specific decoration of CDMs with bioactive molecules deploying metabolic glycoengineering. NIH 3T3 fibroblasts were cultured in the presence of a tetra acetylated azide bearing monosaccharide to metabolically incorporate the synthetic sugar into the glycan structure of extracellular matrix proteins. Glycoengineered CDMs were isolated and analyzed for fibronectin as one of the most abundant ECM species by western blotting and fluorescence labeling of the azide-monosaccharides deploying bioorthogonal chemistries. Glycoengineered CDMs were biocompatible for incorporated and reseeded NIH 3T3 fibroblasts, respectively. Successful modification of glycoengineered CDMs was demonstrated by a therapeutic peptide inhibitor against myostatin. This study details a site-specific, effective, and biocompatible strategy for the decoration of glycan structures within CDMs for future tissue engineering application.