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Surface functionalization of polyurethane scaffolds mimicking the myocardial microenvironment to support cardiac primitive cells.

PloS one (2018-07-07)
Monica Boffito, Franca Di Meglio, Pamela Mozetic, Sara Maria Giannitelli, Irene Carmagnola, Clotilde Castaldo, Daria Nurzynska, Anna Maria Sacco, Rita Miraglia, Stefania Montagnani, Nicoletta Vitale, Mara Brancaccio, Guido Tarone, Francesco Basoli, Alberto Rainer, Marcella Trombetta, Gianluca Ciardelli, Valeria Chiono
ABSTRACT

Scaffolds populated with human cardiac progenitor cells (CPCs) represent a therapeutic opportunity for heart regeneration after myocardial infarction. In this work, square-grid scaffolds are prepared by melt-extrusion additive manufacturing from a polyurethane (PU), further subjected to plasma treatment for acrylic acid surface grafting/polymerization and finally grafted with laminin-1 (PU-LN1) or gelatin (PU-G) by carbodiimide chemistry. LN1 is a cardiac niche extracellular matrix component and plays a key role in heart formation during embryogenesis, while G is a low-cost cell-adhesion protein, here used as a control functionalizing molecule. X-ray photoelectron spectroscopy analysis shows nitrogen percentage increase after functionalization. O1s and C1s core-level spectra and static contact angle measurements show changes associated with successful functionalization. ELISA assay confirms LN1 surface grafting. PU-G and PU-LN1 scaffolds both improve CPC adhesion, but LN1 functionalization is superior in promoting proliferation, protection from apoptosis and expression of differentiation markers for cardiomyocytes, endothelial and smooth muscle cells. PU-LN1 and PU scaffolds are biodegraded into non-cytotoxic residues. Scaffolds subcutaneously implanted in mice evoke weak inflammation and integrate with the host tissue, evidencing a significant blood vessel density around the scaffolds. PU-LN1 scaffolds show their superiority in driving CPC behavior, evidencing their promising role in myocardial regenerative medicine.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
4-Nitro-3-(trifluoromethyl)phenol, 99% (GC)
Sigma-Aldrich
L-Lysine ethyl ester dihydrochloride, ≥99.0% (AT)
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Anti-Rabbit IgG (whole molecule)–Peroxidase antibody produced in goat, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine (TMB) Liquid Substrate System for ELISA, peroxidase substrate
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N-Acetyl-D-lactosamine, ≥98%
Sigma-Aldrich
N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide, ≥97.0% (T)