Direct and Indirect Biomimetic Peptide Modification of Alginate: Efficiency, Side Reactions, and Cell Response.

Anna Golunova,Josef Jaroš,Ognen Pop-Georgievski,Václav Chochola,Zuzana Mikšovská,Aleš Hampl,Nadiia Velychkivska,Vladimír Proks

doi:10.3390/ijms22115731

Abstract

In the fast-developing field of tissue engineering there is a constant demand for new materials as scaffolds for cell seeding, which can better mimic a natural extracellular matrix as well as control cell behavior. Among other materials, polysaccharides are widely used for this purpose. One of the main candidates for scaffold fabrication is alginate. However, it lacks sites for cell adhesion. That is why one of the steps toward the development of suitable scaffolds for cells is the introduction of the biofunctionality to the alginate structure. In this work we focused on bone-sialoprotein derived peptide (TYRAY) conjugation to the molecule of alginate. Here the comparison study on four different approaches of peptide conjugation was performed including traditional and novel modification methods, based on 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxy succinimide (EDC/NHS), 4-(4,6-dimethoxy-1,3,5-triazine-2-yl)-4-methylmorpholinium chloride (DMTMM), thiol-Michael addition and Cu-catalyzed azide–alkyne cycloaddition reactions. It was shown that the combination of the alginate amidation with the use of and subsequent Cu-catalyzed azide–alkyne cycloaddition led to efficient peptide conjugation, which was proven with both NMR and XPS methods. Moreover, the cell culture experiment proved the positive effect of peptide presence on the adhesion of human embryonic stem cells.

Highlights

Tissue engineering is a fast-developing area of research, which requires new materials as scaffolds for cell seeding
It was shown that the combination of the alginate amidation with the use of and subsequent Cu-catalyzed azide–alkyne cycloaddition led to efficient peptide conjugation, which was proven with both NMR and X-Ray Photoelectron Spectroscopy (XPS) methods
Even though human embryonic stem cells (hESC) are very sensitive to the quality and adhesion properties of the microenvironment, our results show that the modified alginates allow functional cell– material interaction for further control of cell fate

Summary

Introduction

Tissue engineering is a fast-developing area of research, which requires new materials as scaffolds for cell seeding. These materials should be biocompatible and give cells seeded on them mechanical support and provide them a diversity of biological signals to mimic natural tissues. Due to their intrinsic biocompatibility, natural polysaccharides are widely used as materials for scaffolds. Alginate is one of the most used for scaffold fabrication. The alginate molecule does not contain sites for cell attachment to provide any cell-specific receptor interactions needed for controlling and amending possible cell responses

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Methods

Results

Conclusion