Dynamic reorganization and enzymatic remodeling of type IV collagen at cell-biomaterial interface

Coelho, N.M., Llopis-Hernandez, V. , Salmeron-Sanchez, M. and Altankov, G. (2016) Dynamic reorganization and enzymatic remodeling of type IV collagen at cell-biomaterial interface. In: Christov, C. Z. (ed.) Insights into Enzyme Mechanisms and Functions from Experimental and Computational Methods. Series: Advances in protein chemistry and structural biology (105). Elsevier, pp. 81-104. ISBN 9780128048252 (doi:10.1016/bs.apcsb.2016.06.001)

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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.

Item Type:Book Sections
Status:Published
Glasgow Author(s) Enlighten ID:Salmeron-Sanchez, Professor Manuel and Llopis-Hernandez, Dr Virginia
Authors: Coelho, N.M., Llopis-Hernandez, V., Salmeron-Sanchez, M., and Altankov, G.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
College of Science and Engineering > School of Engineering > Biomedical Engineering
Journal Name:Advances in protein chemistry and structural biology
Publisher:Elsevier
ISSN:1876-1623
ISBN:9780128048252
Published Online:14 July 2016

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