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.

• Keywords
• NMR, XPS, adhesion-promoting peptide, alginate, cell adhesion, hESC, polysaccharide modification,
• MeSH
• Alginates chemistry   MeSH
• Amines chemistry   MeSH
• Biomimetics * methods   MeSH
• Cell Adhesion MeSH
• Cell Culture Techniques MeSH
• Cell Line MeSH
• Click Chemistry MeSH
• Magnetic Resonance Spectroscopy MeSH
• Molecular Structure MeSH
• Peptides chemistry   pharmacology   MeSH
• Tissue Engineering * methods   MeSH
• Tissue Scaffolds * MeSH
• Cell Survival drug effects   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Alginates MeSH
• Amines MeSH
• Peptides MeSH

Galectin-3 (Gal-3) is a β-galactoside-binding protein that influences various cell functions, including cell adhesion. We focused on the role of Gal-3 as an extracellular ligand mediating cell-matrix adhesion. We used human adipose tissue-derived stem cells and human umbilical vein endothelial cells that are promising for vascular tissue engineering. We found that these cells naturally contained Gal-3 on their surface and inside the cells. Moreover, they were able to associate with exogenous Gal-3 added to the culture medium. This association was reduced with a β-galactoside LacdiNAc (GalNAcβ1,4GlcNAc), a selective ligand of Gal-3, which binds to the carbohydrate recognition domain (CRD) in the Gal-3 molecule. This ligand was also able to detach Gal-3 newly associated with cells but not Gal-3 naturally present on cells. In addition, Gal-3 preadsorbed on plastic surfaces acted as an adhesion ligand for both cell types, and the cell adhesion was resistant to blocking with LacdiNAc. This result suggests that the adhesion was mediated by a binding site different from the CRD. The blocking of integrin adhesion receptors on cells with specific antibodies revealed that the cell adhesion to the preadsorbed Gal-3 was mediated, at least partially, by β1 and αV integrins-namely α5β1, αVβ3, and αVβ1 integrins.

• Keywords
• ADSC, HUVEC, carbohydrate, galectin, integrin,
• MeSH
• Cell Adhesion * MeSH
• Human Umbilical Vein Endothelial Cells cytology   physiology   MeSH
• Galectins metabolism   MeSH
• Integrins metabolism   MeSH
• Blood Proteins metabolism   MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Mesenchymal Stem Cells cytology   physiology   MeSH
• Cell-Matrix Junctions metabolism   MeSH
• Protein Binding MeSH
• Binding Sites MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Galectins MeSH
• Integrins MeSH
• Blood Proteins MeSH
• LGALS3 protein, human MeSH Browser