TY - JOUR
T1 - Extracellular matrix production and regulation in micropatterned endothelial cells
AU - Anderson, Deirdre E.J.
AU - Hinds, Monica T.
N1 - Funding Information:
The authors gratefully acknowledge funding from the American Heart Association grant 09BGIA2260384 . This work was supported by NIH grant OD011092 for the operation of the Oregon National Primate Research Center.
PY - 2012/10/12
Y1 - 2012/10/12
N2 - 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.
AB - 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.
KW - Collagen
KW - Endothelial cells
KW - Fibronectin
KW - Micropattern
KW - TGF-β
KW - TNF
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U2 - 10.1016/j.bbrc.2012.09.034
DO - 10.1016/j.bbrc.2012.09.034
M3 - Article
C2 - 22995321
AN - SCOPUS:84867657188
SN - 0006-291X
VL - 427
SP - 159
EP - 164
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -