A dynamic model of calcific nodule destabilization in response to monocyte- and oxidized lipid-induced matrix metalloproteinases.
Vulnerable plaque remains clinically undetectable, and there is no accepted in vitro model. We characterize the calcific nodules produced by calcifying vascular cells (CVC) in ApoE-null mice, demonstrating increased destabilization of cultured nodules in the presence of oxidized low-density lipoprotein (oxLDL) and monocytes under pulsatile shear stress. CVC implanted in the subcutaneous space of hyperlipidemic mice produced nodules revealing features of calcific atherosclerotic plaque including a fibrous cap, cholesterol clefts, thin shoulder, lipids, and calcium mineral deposits. CVC nodules seeded in the pulsatile flow channel (τ(avg) = 23 dyn/cm(2), ∂τ/∂t = 71 dyn·cm(-2)·s(-1)) underwent deformation and destabilization. Computational fluid dynamics revealed distinct shear force profiles on the nodules. Presence of oxLDL or monocytic THP-1 cells significantly increased the numbers of nodules destabilized from the substrate. Both oxLDL and THP-1 increased matrix metalloproteinase (MMP) activity in CVC. The MMP inhibitor GM6001 significantly reversed oxLDL- and THP-1-induced nodule destabilization, whereas overexpression of MMP-9 increased destabilization. These findings demonstrate that CVC-derived nodules resembled calcific atherosclerotic plaque and were destabilized in the presence of active lipids and monocytes via induction of MMPs.
Dept. of Biomedical Engineering, University of Southern California, Los Angeles, CA 90089, USA.
Rongsong
R
Li
David
D
Mittelstein
Juhyun
J
Lee
Karen
K
Fang
Rohit
R
Majumdar
Yin
Y
Tintut
Linda L
LL
Demer
Tzung K
TK
Hsiai
D000818:Animals; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D002478:Cells, Cultured; D004151:Dipeptides / Q000494:pharmacology; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006952:Hyperlipoproteinemia Type III / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D007150:Immunohistochemistry; D008077:Lipoproteins, LDL / Q000378:metabolism* / Q000494:pharmacology; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D051379:Mice; D018345:Mice, Knockout; D009000:Monocytes / Q000166:cytology / Q000378:metabolism*; D010084:Oxidation-Reduction; D058226:Plaque, Atherosclerotic / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D012212:Rheology; D013314:Stress, Mechanical; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000494:pharmacology; D061205:Vascular Calcification / Q000150:complications / Q000378:metabolism* / Q000473:pathology
D001057:Apolipoproteins E; D004151:Dipeptides; D008077:Lipoproteins, LDL; D061965:Matrix Metalloproteinase Inhibitors; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D019715:Tissue Inhibitor of Metalloproteinase-1; C099252:oxidized low density lipoprotein; D020780:Matrix Metalloproteinase 9