Generation of an azide-modified extracellular matrix by adipose-derived stem cells using metabolic glycoengineering

Svenja Nellinger,Alexander Southan,Valentin Wittmann,Silke Keller,Petra J Kluger,Ann-Cathrin Volz

doi:10.1515/cdbme-2019-0099

Abstract

Abstract Natural extracellular matrix (ECM) represents an ideal biomaterial for tissue engineering and regenerative medicine approaches. For further functionalization, there is a need for specific addressable functional groups within this biomaterial. Metabolic glycoengineering (MGE) provides a technique to incorporate modified monosaccharide derivatives into the ECM during their assembly, which was shown by us earlier for the production of a modified fibroblast-derived dermal ECM. In this study, adipose-derived stem cells (ASCs) were treated with the azide-modified monosaccharide derivate 1,3,4,6-tetra-O-acetyl-N-azidoacetylgalactosamine (Ac4GalNAz). Toxicity and viability assays after 24 h and 72 h incubation revealed high biocompatibility of Ac4GalNAz in contact with ASCs. The successful incorporation of the functional azide groups into the glycocalyx and the ECM of the ASCs was proven by conjugation with a fluorescent dye via a copper-catalyzed click reaction. Thus, Ac4GalNAz in combination with ASCs was confirmed to achieve an azidemodified ECM as a multifunctional biomaterial for further applications.

Highlights

The extracellular matrix (ECM) represents the natural environment of cells in an organism, wherein it is synthesized and assembled by tissue-specific cells
We employed metabolic glycoengineering (MGE) with the azide-modified monosaccharide derivate 1,3,4,6-tetra-Oacetyl-N-azidoacetylgalactosamine (Ac4GalNAz) to generate a human fibroblast-derived dermal ECM containing azide groups [11] that can be addressed by copper-catalyzed azidealkyne cycloaddition [12, 13]
We aimed to prove whether the glycocalyx and ECM of adipose-derived stem cells (ASCs) can be modified with azide groups using Ac4GalNAz in MGE (Figure 1)

Summary

Introduction

The extracellular matrix (ECM) represents the natural environment of cells in an organism, wherein it is synthesized and assembled by tissue-specific cells. Chemical modification of ECM compounds is challenging as it might affect matrix integrity To address these issues, metabolic glycoengineering (MGE) represents a promising tool. Metabolic glycoengineering (MGE) represents a promising tool This method is based on the incorporation of chemically modified monosaccharide derivatives into the natural intra- and extracellular oligosaccharide structures of the cell by its natural metabolic pathways [7, 8]. We employed MGE with the azide-modified monosaccharide derivate 1,3,4,6-tetra-Oacetyl-N-azidoacetylgalactosamine (Ac4GalNAz) to generate a human fibroblast-derived dermal ECM containing azide groups [11] that can be addressed by copper-catalyzed azidealkyne cycloaddition [12, 13]. Gutmann et al reported a similar approach using the corresponding azide-modified glucosamine derivative [14]

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Methods

Conclusion