title: Polyelectrolyte multilayer films allow seeded human progenitor-derived endothelial cells to remain functional under shear stress in vitro.
abstract: There is considerable interest in making multilayer films for various applications, among which are cell contacting biomaterials, allowing new opportunities to prepare functionalized biomaterials. In this study we have explored the capability of poly(sodium-4-styrene sulfonate)/poly(allylamine hydrochloride) polyelectrolyte multilayer films (PMFs) as functional coatings for human progenitor-derived endothelial cells (PDECs), since the latter are a potential source of endothelial-type cells to be used in bioartificial vascular substitutes. We performed investigations with PDECs derived from peripheral blood and characterized as endothelial cells. After forming a confluent monolayer on PMFs they were exposed to laminar pulsatile physiological shear stress. We investigated whether PDECs were able to withstand shear stress and to respond at the mRNA (microarray analysis) and protein levels (thrombomodulin and tissue factor functional activity), in comparison with collagen I and fibrin glue used as controls. After shear stress the PDECs remained spread on the substrates, with a resulting increase in the number of expressed genes. Considering the functional significance of our findings for the regulation of coagulation and fibrinolytic factors, mRNA tissue plasminogen activator and thrombomodulin, profibrinolytic and thrombin inhibiting respectively, were overexpressed in PDECs after 6h shear stress. von Willebrand factor showed down-regulation, while tissue factor was up-regulated. We can speculate that PMFs could favour anti-thrombogenic activity by PDECs because activated protein C generation, measuring thrombomodulin activity, was particularly high on PMFs, but unchanged after 6h shear stress. Thus, PMFs could represent suitable coatings able to provide functional surfaces for endothelialization with PDECs.
authors.affiliation: Inserm U577, Bordeaux and Université Victor Segalen Bordeaux 2 F-33076, France.
authors.forename: N B
authors.initials: NB
authors.lastname: Thébaud
authors.forename: R
authors.initials: R
authors.lastname: Bareille
authors.forename: R
authors.initials: R
authors.lastname: Daculsi
authors.forename: Ch
authors.initials: Ch
authors.lastname: Bourget
authors.forename: M
authors.initials: M
authors.lastname: Rémy
authors.forename: H
authors.initials: H
authors.lastname: Kerdjoudj
authors.forename: P
authors.initials: P
authors.lastname: Menu
authors.forename: L
authors.initials: L
authors.lastname: Bordenave
mesh_terms: D000818:Animals; D024042:Collagen Type I / Q000494:pharmacology; D004573:Electrolytes / Q000494:pharmacology*; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D013234:Stem Cells / Q000166:cytology*; D013314:Stress, Mechanical; D018180:Thrombomodulin / Q000235:genetics / Q000378:metabolism; D013925:Thromboplastin / Q000235:genetics / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D004573:Electrolytes; D018180:Thrombomodulin; D013925:Thromboplastin