Interstitial flow promotes vascular fibroblast, myofibroblast, and smooth muscle cell motility in 3-D collagen I via upregulation of MMP-1.

Neointima formation often occurs in regions where the endothelium has been damaged and the transmural interstitial flow is elevated. Vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts (FBs/MFBs) contribute to intimal thickening by migrating from the media and adventitia into the site of injury. In this study, for the first time, the direct effects of interstitial flow on SMC and FB/MFB migration were investigated in an in vitro three-dimensional system. Collagen I gels were used to mimic three-dimensional extracellular matrix (ECM) for rat aortic SMCs and FBs/MFBs. Exposure to interstitial flow induced by 1 cmH(2)O pressure differential (shear stress, approximately 0.05 dyn/cm(2); flow velocity, approximately 0.5 microm/s; and Darcy permeability, approximately 10(-11) cm(2)) substantially enhanced cell motility. Matrix metalloproteinase (MMP) inhibitor (GM-6001) abolished flow-induced migration augmentation, which suggested that the enhanced motility was MMP dependent. The upregulation of MMP-1 played a critical role for the flow-enhanced motility, which was further confirmed by silencing MMP-1 gene expression. Longer exposures to higher flows suppressed the number of migrated cells, although MMP-1 gene expression remained high. This suppression was a result of both flow-induced tissue inhibitor of metalloproteinase-1 upregulation and increased apoptotic and necrotic cell death. Interstitial flow did not affect MMP-2 gene expression or activity in the collagen I gel for any cell type. Our findings shed light on the mechanism by which vascular SMCs and FBs/MFBs contribute to intimal thickening in regions of vascular injury where interstitial flow is elevated.

[1]  A. Remuzzi,et al.  Shear stress downregulation of platelet-derived growth factor receptor-beta and matrix metalloprotease-2 is associated with inhibition of smooth muscle cell invasion and migration. , 2000, Circulation.

[2]  Eben Alsberg,et al.  FRET measurements of cell-traction forces and nano-scale clustering of adhesion ligands varied by substrate stiffness. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Jean-Baptiste Michel,et al.  Topological Determinants and Consequences of Adventitial Responses to Arterial Wall Injury , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[4]  S. Chien,et al.  Tumor cell cycle arrest induced by shear stress: Roles of integrins and Smad , 2008, Proceedings of the National Academy of Sciences.

[5]  M. Humbert,et al.  Cellular and molecular pathobiology of pulmonary arterial hypertension. , 2004, Journal of the American College of Cardiology.

[6]  P. Lin,et al.  Chlorotyrosine promotes human aortic smooth muscle cell migration through increasing superoxide anion production and ERK1/2 activation. , 2008, Atherosclerosis.

[7]  G. Karakiulakis,et al.  Cell Type-specific Effect of Hypoxia and Platelet-derived Growth Factor-BB on Extracellular Matrix Turnover and Its Consequences for Lung Remodeling* , 2007, Journal of Biological Chemistry.

[8]  P. Serruys,et al.  Proliferation and extracellular matrix synthesis of smooth muscle cells cultured from human coronary atherosclerotic and restenotic lesions. , 1994, Journal of the American College of Cardiology.

[9]  D. Lauffenburger,et al.  Migration of tumor cells in 3D matrices is governed by matrix stiffness along with cell-matrix adhesion and proteolysis. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[10]  R. Wilensky,et al.  Effect of atherosclerosis on transmural convection an arterial ultrastructure. Implications for local intravascular drug delivery. , 1997, Arteriosclerosis, thrombosis, and vascular biology.

[11]  M. Swartz,et al.  Fibroblast alignment under interstitial fluid flow using a novel 3-D tissue culture model. , 2003, American journal of physiology. Heart and circulatory physiology.

[12]  J. Tarbell,et al.  Flow through internal elastic lamina affects shear stress on smooth muscle cells (3D simulations). , 2002, American journal of physiology. Heart and circulatory physiology.

[13]  J. Levick.  Flow through interstitium and other fibrous matrices. , 1987, Quarterly journal of experimental physiology.

[14]  Z. Galis,et al.  This Review Is Part of a Thematic Series on Matrix Metalloproteinases, Which Includes the following Articles: Matrix Metalloproteinase Inhibition after Myocardial Infarction: a New Approach to Prevent Heart Failure? Matrix Metalloproteinases in Vascular Remodeling and Atherogenesis: the Good, the Ba , 2022 .

[15]  R. Kaunas,et al.  Fluid shear stress modulates endothelial cell invasion into three-dimensional collagen matrices. , 2008, American journal of physiology. Heart and circulatory physiology.

[16]  A. Zalewski,et al.  Role of matrix metalloproteinases and their tissue inhibitors in the regulation of coronary cell migration. , 1999, Arteriosclerosis, thrombosis, and vascular biology.

[17]  Saroja Ramanujan,et al.  Diffusion and convection in collagen gels: implications for transport in the tumor interstitium. , 2002, Biophysical journal.

[18]  P. Weissberg,et al.  Migration of adventitial myofibroblasts following vascular balloon injury: insights from in vivo gene transfer to rat carotid arteries. , 2003, Cardiovascular research.

[19]  D. Rumschitzki,et al.  Transport in rat vessel walls. I. Hydraulic conductivities of the aorta, pulmonary artery, and inferior vena cava with intact and denuded endothelia. , 2006, American journal of physiology. Heart and circulatory physiology.

[20]  P. Carmeliet,et al.  Temporal and Topographic Matrix Metalloproteinase Expression after Vascular Injury in Mice , 1999, Thrombosis and Haemostasis.

[21]  M. Ringuette,et al.  Collagen I but not Matrigel matrices provide an MMP-dependent barrier to ovarian cancer cell penetration , 2008, BMC Cancer.

[22]  C. Brinckerhoff,et al.  Induction of collagenase and stromelysin gene expression by mechanical injury in a vascular smooth muscle‐derived cell line , 1993, Journal of cellular physiology.

[23]  K. Preissner,et al.  The FASEB Journal • Research Communication Calpain counteracts mechanosensitive apoptosis of vascular smooth muscle cells in vitro and in vivo , 2022 .

[24]  Douglas Lauffenburger,et al.  Interstitial fluid flow intensity modulates endothelial sprouting in restricted Src-activated cell clusters during capillary morphogenesis. , 2009, Tissue engineering. Part A.

[25]  K Walsh,et al.  Evidence for the rapid onset of apoptosis in medial smooth muscle cells after balloon injury. , 1997, Circulation.

[26]  A. Newby,et al.  Matrix metalloproteinases regulate migration, proliferation, and death of vascular smooth muscle cells by degrading matrix and non-matrix substrates. , 2006, Cardiovascular research.

[27]  J. Wilcox,et al.  Identification of a potential role for the adventitia in vascular lesion formation after balloon overstretch injury of porcine coronary arteries. , 1996, Circulation.

[28]  A. Chiavegato,et al.  Contribution of Adventitial Fibroblasts to Neointima Formation and Vascular Remodeling: From Innocent Bystander to Active Participant , 2001, Circulation research.

[29]  L. Griffith,et al.  Capturing complex 3D tissue physiology in vitro , 2006, Nature Reviews Molecular Cell Biology.

[30]  John A. Pedersen,et al.  Effects of extracellular fiber architecture on cell membrane shear stress in a 3D fibrous matrix. , 2007, Journal of biomechanics.

[31]  J. Tarbell.  Mass transport in arteries and the localization of atherosclerosis. , 2003, Annual review of biomedical engineering.

[32]  M. Reidy,et al.  Matrix metalloproteinases of vascular wall cells are increased in balloon-injured rat carotid artery. , 1994, Journal of vascular surgery.

[33]  John M. Tarbell,et al.  Effect of Fluid Flow on Smooth Muscle Cells in a 3-Dimensional Collagen Gel Model , 2000, Arteriosclerosis, thrombosis, and vascular biology.

[34]  James F Glockner,et al.  Increased shear stress with upregulation of VEGF-A and its receptors and MMP-2, MMP-9, and TIMP-1 in venous stenosis of hemodialysis grafts. , 2008, American journal of physiology. Heart and circulatory physiology.

[35]  M. Swartz,et al.  Interstitial flow and its effects in soft tissues. , 2007, Annual review of biomedical engineering.

[36]  J. Hoff,et al.  Zymographic techniques for the analysis of matrix metalloproteinases and their inhibitors , 2005 .

[37]  J. Raffetto,et al.  Matrix metalloproteinases and their inhibitors in vascular remodeling and vascular disease. , 2008, Biochemical pharmacology.

[38]  A. Zalewski,et al.  Adventitial myofibroblasts contribute to neointimal formation in injured porcine coronary arteries. , 1996, Circulation.

[39]  John M Tarbell,et al.  Effects of fluid shear stress on adventitial fibroblast migration: implications for flow-mediated mechanisms of arterialization and intimal hyperplasia. , 2007, American journal of physiology. Heart and circulatory physiology.

[40]  M. Reidy,et al.  Smooth muscle cell migration and matrix metalloproteinase expression after arterial injury in the rat. , 1994, Circulation research.

[41]  E. Schiffrin,et al.  Apoptosis in vasculature of spontaneously hypertensive rats: effect of an angiotensin converting enzyme inhibitor and a calcium channel antagonist. , 1998, American journal of hypertension.

[42]  Y. Fung,et al.  Cellular and molecular mechanisms of pulmonary vascular remodeling: role in the development of pulmonary hypertension. , 2004, Microvascular research.

[43]  A. Newby,et al.  Dual role of matrix metalloproteinases (matrixins) in intimal thickening and atherosclerotic plaque rupture. , 2005, Physiological reviews.

[44]  Shu Chien,et al.  Biochemistry and biomechanics of cell motility. , 2005, Annual review of biomedical engineering.

[45]  John M Tarbell,et al.  Shear stress inhibits smooth muscle cell migration via nitric oxide-mediated downregulation of matrix metalloproteinase-2 activity. , 2005, American journal of physiology. Heart and circulatory physiology.

[46]  T. Henry,et al.  Pathophysiology of coronary artery restenosis. , 2002, Reviews in cardiovascular medicine.

[47]  W. Hornebeck,et al.  Collagen zymography as a sensitive and specific technique for the determination of subpicogram levels of interstitial collagenase. , 1998, Analytical biochemistry.

[48]  M. Lever,et al.  Filtration through damaged and undamaged rabbit thoracic aorta. , 1984, The American journal of physiology.

[49]  A. Tedgui,et al.  Apoptosis in the vasculature: mechanisms and functional importance , 2000, British journal of pharmacology.

[50]  J. Tarbell,et al.  Interstitial flow through the internal elastic lamina affects shear stress on arterial smooth muscle cells. , 2000, American journal of physiology. Heart and circulatory physiology.

[51]  M. Morimatsu,et al.  Role of Rho-Associated Kinase in Neointima Formation After Vascular Injury , 2001, Circulation.

[52]  G. A. Limb,et al.  Human corneal epithelial cells require MMP-1 for HGF-mediated migration on collagen I. , 2003, Investigative ophthalmology & visual science.

[53]  J. W. Von den Hoff,et al.  Zymographic techniques for the analysis of matrix metalloproteinases and their inhibitors. , 2005, BioTechniques.

[54]  B. Shepherd,et al.  Laminar shear stress stimulates vascular smooth muscle cell apoptosis via the Akt pathway , 2008, Journal of cellular physiology.

[55]  J. Tarbell,et al.  Modeling interstitial flow in an artery wall allows estimation of wall shear stress on smooth muscle cells. , 1995, Journal of biomechanical engineering.