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  • Fibronectin Adsorption on Electrospun Synthetic Vascular Grafts Attracts Endothelial Progenitor Cells and Promotes Endothelialization in Dynamic In Vitro Culture.

Fibronectin Adsorption on Electrospun Synthetic Vascular Grafts Attracts Endothelial Progenitor Cells and Promotes Endothelialization in Dynamic In Vitro Culture.

Cells (2020-03-27)
Ruben Daum, Dmitri Visser, Constanze Wild, Larysa Kutuzova, Maria Schneider, Günter Lorenz, Martin Weiss, Svenja Hinderer, Ulrich A Stock, Martina Seifert, Katja Schenke-Layland
ABSTRACT

Appropriate mechanical properties and fast endothelialization of synthetic grafts are key to ensure long-term functionality of implants. We used a newly developed biostable polyurethane elastomer (TPCU) to engineer electrospun vascular scaffolds with promising mechanical properties (E-modulus: 4.8 ± 0.6 MPa, burst pressure: 3326 ± 78 mmHg), which were biofunctionalized with fibronectin (FN) and decorin (DCN). Neither uncoated nor biofunctionalized TPCU scaffolds induced major adverse immune responses except for minor signs of polymorph nuclear cell activation. The in vivo endothelial progenitor cell homing potential of the biofunctionalized scaffolds was simulated in vitro by attracting endothelial colony-forming cells (ECFCs). Although DCN coating did attract ECFCs in combination with FN (FN + DCN), DCN-coated TPCU scaffolds showed a cell-repellent effect in the absence of FN. In a tissue-engineering approach, the electrospun and biofunctionalized tubular grafts were cultured with primary-isolated vascular endothelial cells in a custom-made bioreactor under dynamic conditions with the aim to engineer an advanced therapy medicinal product. Both FN and FN + DCN functionalization supported the formation of a confluent and functional endothelial layer.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Fibronectin human plasma, lyophilized powder, BioReagent, suitable for cell culture, ≥95% (SDS-PAGE)
Sigma-Aldrich
Human Serum, from human male AB plasma, USA origin, sterile-filtered
Sigma-Aldrich
Anti-Fibronectin antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Thiazolyl Blue Tetrazolium Bromide, 98%
Sigma-Aldrich
Paraformaldehyde, reagent grade, crystalline
Sigma-Aldrich
1,1,1,3,3,3-Hexafluoro-2-propanol, for synthesis