title: Microcapillary-like structures prompted by phospholipase A2 activation in endothelial cells and pericytes co-cultures on a polyhydroxymethylsiloxane thin film.
abstract: A thin film of poly(hydroxymethylsiloxane) (PHMS) has been deposited on glass dishes and tested as artificial support material for vascularization from mixed cultures of endothelial cells (EC) and pericytes (PC). The EC/PC co-cultures adhered massively on PHMS, with the formation of net-like microcapillary structures. Such evidence was not found on control glass substrates in the same co-culture conditions neither on PHMS for EC and PC in monocultures. The physicochemical characterization of PHMS and control glass surface by time-of-flight secondary ion mass spectrometry, X-ray photoelectron spectroscopy, water contact angle and atomic force microscopy, pointed to the main role of the polymer hydrophobilicy to explain the observed cellular behavior. Moreover, enhanced intercellular cross-talk was evidenced by the up-regulation and activation of cytoplasmic and Ca(2+)-independent phospholipase A(2) (cPLA(2) and iPLA(2)) expression and cPLA(2) phosphorylation, leading to the cell proliferation and microcapillary formation on the PHMS surface, as evidenced by confocal microscopy analyses. Co-cultures, established with growth-arrested PCs by treatment with mitomycin C, showed an increase in EC proliferation on PHMS. AACOCF(3) or co-transfection with cPLA(2) and iPLA(2)siRNA reduced cell proliferation. The results highlight the major role played by EC/PC cross-talk as well as the hydrophobic character of the substrate surface, to promote microcapillary formation. Our findings suggest an attractive strategy for vascular tissue engineering and provide new details on the interplay of artificial substrates and capillary formation.
authors.affiliation: Department of Clinical and Molecular Biomedicine, University of Catania, viale Andrea Doria 5, 95125 Catania, Italy.
authors.forename: Carmelina Daniela
authors.initials: CD
authors.lastname: Anfuso
authors.forename: Carla
authors.initials: C
authors.lastname: Motta
authors.forename: Cristina
authors.initials: C
authors.lastname: Satriano
authors.forename: Salvatore
authors.initials: S
authors.lastname: Gennaro
authors.forename: Giovanni
authors.initials: G
authors.lastname: Marletta
authors.forename: Giovanni
authors.initials: G
authors.lastname: Giurdanella
authors.forename: Mario
authors.initials: M
authors.lastname: Alberghina
authors.forename: Gabriella
authors.initials: G
authors.lastname: Lupo
mesh_terms: D000818:Animals; D040761:Biomimetic Materials / Q000737:chemistry / Q000494:pharmacology*; D002196:Capillaries / Q000166:cytology*; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D003593:Cytoplasm / Q000201:enzymology; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005898:Glass / Q000737:chemistry; D020286:Pericytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D054467:Phospholipases A2 / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D012833:Siloxanes / Q000737:chemistry / Q000494:pharmacology*; D017155:Wettability
chemical_list: D012333:RNA, Messenger; D012833:Siloxanes; C515054:poly(hydroxymethylsiloxane); D054467:Phospholipases A2