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.

• Klíčová slova
• NMR, XPS, adhesion-promoting peptide, alginate, cell adhesion, hESC, polysaccharide modification,
• MeSH
• algináty chemie   MeSH
• aminy chemie   MeSH
• biomimetika * metody   MeSH
• buněčná adheze MeSH
• buněčné kultury MeSH
• buněčné linie MeSH
• magnetická rezonanční spektroskopie MeSH
• molekulární struktura MeSH
• peptidy chemie   farmakologie   MeSH
• syntetická chemie okamžité shody MeSH
• tkáňové inženýrství * metody   MeSH
• tkáňové podpůrné struktury * MeSH
• viabilita buněk účinky léků   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• algináty MeSH
• aminy MeSH
• peptidy 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.

• Klíčová slova
• ADSC, HUVEC, carbohydrate, galectin, integrin,
• MeSH
• buněčná adheze * MeSH
• endoteliální buňky pupečníkové žíly (lidské) cytologie   fyziologie   MeSH
• galektiny metabolismus   MeSH
• integriny metabolismus   MeSH
• krevní proteiny metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   fyziologie   MeSH
• spoje buňka-matrix metabolismus   MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• galektiny MeSH
• integriny MeSH
• krevní proteiny MeSH
• LGALS3 protein, human MeSH Prohlížeč