title: Extracellular matrix production and regulation in micropatterned endothelial cells.
abstract: Production and maintenance of extracellular matrix (ECM) is an essential aspect of endothelial cell (EC) function. ECM surfaces composed of collagen type IV and laminin support an atheroprotective endothelium, while fibronectin may encourage an atheroprone endothelium through inflammation or wound repair signaling. ECs maintain this underlying structure through regulation of protein production and degradation, yet the role of cytoskeletal alignment on this regulation is unknown. To examine the regulation and production of ECM by ECs with an atheroprotective phenotype, ECs were micropatterned onto lanes, which created an elongated EC morphology similar to that seen with unidirectional fluid shear stress application. Collagen IV and fibronectin protein production were measured as were gene expression of collagen IV, fibronectin, laminin, MMP2, MMP9, TIMP1, TIMP2, and TGF-β1. ECs were also treated with TNF to simulate an injury model. Micropattern-induced elongation led to significant increases in collagen IV and fibronectin protein production, and collagen IV, laminin, and TGF-β1 gene expression, but no significant changes in the MMP or TIMP genes. TNF treatment significantly increased collagen IV gene and protein production. These results suggest that the increase in ECM synthesis in micropattern-elongated ECs is likely regulated with TGF-β1, and this increase in ECM could be relevant to the atheroprotection needed for maintenance of a healthy endothelium in vivo.
authors.affiliation: Department of Biomedical Engineering, Oregon Health & Science University, CH13B, 3303 SW Bond Avenue, Portland, OR 97239, USA.
authors.forename: Deirdre E J
authors.initials: DE
authors.lastname: Anderson
authors.forename: Monica T
authors.initials: MT
authors.lastname: Hinds
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000096:biosynthesis / Q000235:genetics; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005353:Fibronectins / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D007797:Laminin / Q000096:biosynthesis / Q000235:genetics; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis / Q000235:genetics; D010215:Papio; D014176:Protein Biosynthesis; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis / Q000235:genetics; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000096:biosynthesis / Q000235:genetics; D053773:Transforming Growth Factor beta1 / Q000096:biosynthesis / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D024141:Collagen Type IV; D005353:Fibronectins; D007797:Laminin; D019715:Tissue Inhibitor of Metalloproteinase-1; D053773:Transforming Growth Factor beta1; D014409:Tumor Necrosis Factor-alpha; D019716:Tissue Inhibitor of Metalloproteinase-2; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9