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

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.

Eclectic characterisation of chemically modified cell-derived matrices obtained by metabolic glycoengineering and re-assessment of commonly used methods

“…the outstanding bioactive properties) with abiotic functional groups which are a powerful tool for (bio) conjugation. 10,[17][18][19][20] Characterisation of the successful azide modication of cell-derived ECM via MGE was successfully completed by covalently linking clickECM azides to an alkynecoupled uorophore via Huisgen 1,3-dipolar azide-alkyne cycloaddition 10,18-20 (ESI Fig. 1 †).…”

“…[13][14][15][16][17] Using the same monosaccharide but adipose-derived stem cells (ASCs) instead of broblasts, Nellinger et al demonstrated that this method can be used to generate azide-modied tissue-specic ECM. 18 As an alternative to the above mentioned Ac 4 -GalNAz, Gutmann et al demonstrated the successful modication of NIH-3T3 broblasts-derived ECM using the glucosamine derivate 2-azidoacetylamino-2-deoxy-(1,3,4,6)tetra-O-acetyl-D-glucopyranoside (Ac 4 GlcNAz). 19,20 Despite the successful modication of ECM with azides via MGE and their demonstrated applicability as surface coatings or bioconjugation platforms, 10,17,19 it is not yet understood if MGE changes properties of azide-modied ECM (e.g.…”

Azide-Functional Extracellular Matrix Coatings as a Bioactive Platform for Bioconjugation

“…Biomaterials, in general, attempt to mimic these multifactorial aspects of ECM function; however, synthetic materials and single ECM components fail to achieve the molecular complexity and organization of the ECM in vivo. , Therefore, the use of complex CDMs as biomaterialsin addition to decellularization of whole tissues and organs , has recently gained attention because they promise to preserve, at least to a large extent, the complexity of native tissue matrices. ,,, However, these biological matrices still lack specific addressable functional groups, which are often required for their use as a biomaterial. , …”

“…Later, Gutmann et al used a similar method to successfully modify the ECM of NIH3T3 fibroblasts using the glucosamine derivate 2-azidoacetylamino-2-deoxy-(1,3,4,6)-tetra- O -acetyl- d -glucopyranoside (Ac 4 GlcNAz) . More recently, Nellinger et al modified the ECM from adipose-derived stem cells (ASCs) with azides through MGE using the above-mentioned Ac 4 GalNAz …”

An Advanced ‘clickECM’ That Can be Modified by the Inverse‐Electron‐Demand Diels‐Alder Reaction

“…Previously, we [9] and others [10] reported the preparation of a functionalized ECM by MGE. For example, we demonstrated the incorporation of azide functionalities into the ECM of human fibroblasts [9a] and ASCs, [9b] which can be addressed via bioorthogonal copper‐catalyzed azide‐alkyne cycloaddition (CuAAC) [11] . We demonstrated different possible applications of the azide‐modified ‘clickECM’ and showed, for example, the positive effect of a clickECM coating on the fibroblast culture and demonstrated the clickECM as a bioconjugation platform using biotin‐streptavidin interaction [12] …”