title: Membrane-type matrix metalloproteinase-mediated angiogenesis in a fibrin-collagen matrix.
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.
authors.affiliation: Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
authors.forename: Annemie
authors.initials: A
authors.lastname: Collen
authors.forename: Roeland
authors.initials: R
authors.lastname: Hanemaaijer
authors.forename: Florea
authors.initials: F
authors.lastname: Lupu
authors.forename: Paul H A
authors.initials: PH
authors.lastname: Quax
authors.forename: Natascha
authors.initials: N
authors.lastname: van Lent
authors.forename: Jos
authors.initials: J
authors.lastname: Grimbergen
authors.forename: Erna
authors.initials: E
authors.lastname: Peters
authors.forename: Pieter
authors.initials: P
authors.lastname: Koolwijk
authors.forename: Victor W M
authors.initials: VW
authors.lastname: van Hinsbergh
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D007611:Aprotinin / Q000494:pharmacology; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects; D003094:Collagen; D003470:Culture Media / Q000494:pharmacology; D003673:Defective Viruses / Q000235:genetics; D004357:Drug Synergism; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D005337:Fibrin; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005822:Genetic Vectors / Q000235:genetics; D006331:Heart Diseases / Q000473:pathology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D008222:Lymphokines / Q000494:pharmacology; D020781:Matrix Metalloproteinase 1 / Q000502:physiology*; D020778:Matrix Metalloproteinase 2 / Q000502:physiology*; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000502:physiology*; D009389:Neovascularization, Pathologic / Q000201:enzymology* / Q000503:physiopathology; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D013876:Thiophenes / Q000494:pharmacology; D013927:Thrombosis / Q000473:pathology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics / Q000502:physiology; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics / Q000502:physiology; D014161:Transduction, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D003470:Culture Media; D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D011480:Protease Inhibitors; D013876:Thiophenes; D019715:Tissue Inhibitor of Metalloproteinase-1; D019717:Tissue Inhibitor of Metalloproteinase-3; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D016222:Fibroblast Growth Factor 2; D010649:Phenylalanine; D005337:Fibrin; D003094:Collagen; D007611:Aprotinin; C080985:batimastat; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1