title: Dynamic Reorganization and Enzymatic Remodeling of Type IV Collagen at Cell-Biomaterial Interface.
abstract: Vascular basement membrane remodeling involves assembly and degradation of its main constituents, type IV collagen (Col IV) and laminin, which is critical during development, angiogenesis, and tissue repair. Remodeling can also occur at cell-biomaterials interface altering significantly the biocompatibility of implants. Here we describe the fate of adsorbed Col IV in contact with endothelial cells adhering on positively charged NH2 or hydrophobic CH3 substrata, both based on self-assembly monolayers (SAMs) and studied alone or mixed in different proportions. AFM studies revealed distinct pattern of adsorbed Col IV, varying from single molecular deposition on pure NH2 to network-like assembly on mixed SAMs, turning to big globular aggregates on bare CH3. Human umbilical endothelial cells (HUVECs) interact better with Col IV adsorbed as single molecules on NH2 surface and readily rearrange it in fibril-like pattern that coincide with secreted fibronectin fibrils. The cells show flattened morphology and well-developed focal adhesion complexes that are rich on phosphorylated FAK while expressing markedly low pericellular proteolytic activity. Conversely, on hydrophobic CH3 substrata HUVECs showed abrogated spreading and FAK phosphorylation, combined with less reorganization of the aggregated Col IV and significantly increased proteolytic activity. The later involves both MMP-2 and MMP-9, as measured by zymography and FITC-Col IV release. The mixed SAMs support intermediate remodeling activity. Taken together these results show that chemical functionalization combined with Col IV preadsorption provides a tool for guiding the endothelial cells behavior and pericellular proteolytic activity, events that strongly affect the fate of cardiovascular implants.
authors: Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain.
authors: N M
authors: NM
authors: Coelho
authors: Center for Biomaterials and Tissue Engineering, Universidad Politécnica de Valencia, Valencia, Spain; School of Engineering, University of Glasgow, Glasgow, United Kingdom.
authors: V
authors: V
authors: Llopis-Hernández
authors: School of Engineering, University of Glasgow, Glasgow, United Kingdom.
authors: M
authors: M
authors: Salmerón-Sánchez
authors: Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain; Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain. Electronic address: altankov@abv.bg.
authors: G
authors: G
authors: Altankov
mesh_terms: D001672:Biocompatible Materials; D024141:Collagen Type IV / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D013499:Surface Properties
chemical_list: D001672:Biocompatible Materials; D024141:Collagen Type IV
keywords: Adsorption; CH(3) and NH(2) groups; Pericellular proteolysis; Remodeling; Reorganization; Self-assembly monolayers; Substratum chemistry; Type IV collagen