Abstract
Adult angiogenesis, associated with pathologic conditions, is often accompanied by the formation of a fibrinous exudate. This temporary matrix consists mainly of fibrin but is intermingled with plasma proteins and collagen fibers. The formation of capillary structures in a fibrinous matrix in vivo was mimicked by an in vitro model, in which human microvascular endothelial cells (hMVECs) seeded on top of a fibrin-10% collagen matrix form capillarylike tubular structures after stimulation with basic fibroblast growth factor/tumor necrosis factor alpha (bFGF/TNF-alpha) or vascular endothelial growth factor (VEGF)/TNF-alpha. In the fibrin-collagen matrix the metalloproteinase inhibitor BB94 inhibited tubule formation by 70% to 80%. Simultaneous inhibition of plasmin and metalloproteinases by aprotinin and BB94 caused a nearly complete inhibition of tubule formation. Adenoviral transduction of tissue inhibitor of metalloproteinases 1 (TIMP-1) and TIMP-3 into endothelial cells revealed that TIMP-3 markedly inhibited angiogenesis, whereas TIMP-1 had only a minor effect. Immunohistochemical analysis showed the presence of matrix metalloproteinase 1 (MMP-1), MMP-2, and membrane-type 1 (MT1)-MMP, whereas MMP-9 was absent. The endothelial production of these MMPs was confirmed by antigen assays and real-time polymerase chain reaction (PCR). MT1-MMP mRNA was markedly increased in endothelial cells under conditions that induced tubular structures. The presence of MMP-1, MMP-2, and MT1-MMP was also demonstrated in vivo in the newly formed vessels of a recanalized arterial mural thrombus. These data suggest that MMPs, in particular MT-MMPs, play a pivotal role in the formation of capillarylike tubular structures in a collagen-containing fibrin matrix in vitro and may be involved in angiogenesis in a fibrinous exudate in vivo.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenoviridae / genetics
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Aprotinin / pharmacology
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Cells, Cultured / cytology
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Cells, Cultured / drug effects
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Collagen
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Culture Media / pharmacology
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Defective Viruses / genetics
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Drug Synergism
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Endothelial Growth Factors / pharmacology
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Endothelium, Vascular / cytology
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Endothelium, Vascular / drug effects
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Fibrin
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Fibroblast Growth Factor 2 / pharmacology
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Genetic Vectors / genetics
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Heart Diseases / pathology
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Humans
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Intercellular Signaling Peptides and Proteins / pharmacology
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Lymphokines / pharmacology
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Matrix Metalloproteinase 1 / physiology*
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Matrix Metalloproteinase 2 / physiology*
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Matrix Metalloproteinases, Membrane-Associated
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Metalloendopeptidases / physiology*
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Neovascularization, Pathologic / enzymology*
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Neovascularization, Pathologic / physiopathology
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Neovascularization, Physiologic / physiology*
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Phenylalanine / analogs & derivatives*
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Phenylalanine / pharmacology
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Protease Inhibitors / pharmacology
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Thiophenes / pharmacology
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Thrombosis / pathology
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Tissue Inhibitor of Metalloproteinase-1 / genetics
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Tissue Inhibitor of Metalloproteinase-1 / physiology
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Tissue Inhibitor of Metalloproteinase-3 / genetics
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Tissue Inhibitor of Metalloproteinase-3 / physiology
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Transduction, Genetic
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Tumor Necrosis Factor-alpha / pharmacology
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Vascular Endothelial Growth Factor A
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Vascular Endothelial Growth Factors
Substances
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Culture Media
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Endothelial Growth Factors
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Intercellular Signaling Peptides and Proteins
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Lymphokines
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Protease Inhibitors
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Thiophenes
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Tissue Inhibitor of Metalloproteinase-1
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Tissue Inhibitor of Metalloproteinase-3
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Tumor Necrosis Factor-alpha
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Vascular Endothelial Growth Factor A
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Vascular Endothelial Growth Factors
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Fibroblast Growth Factor 2
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Phenylalanine
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Fibrin
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Collagen
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Aprotinin
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batimastat
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Matrix Metalloproteinases, Membrane-Associated
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Metalloendopeptidases
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Matrix Metalloproteinase 2
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Matrix Metalloproteinase 1