title: Fibronectin Deposition Participates in Extracellular Matrix Assembly and Vascular Morphogenesis.
abstract: The extracellular matrix (ECM) has been demonstrated to facilitate angiogenesis. In particular, fibronectin has been documented to activate endothelial cells, resulting in their transition from a quiescent state to an active state in which the cells exhibit enhanced migration and proliferation. The goal of this study is to examine the role of polymerized fibronectin during vascular tubulogenesis using a 3 dimensional (3D) cell-derived de-cellularized matrix. A fibronectin-rich 3D de-cellularized ECM was used as a scaffold to study vascular morphogenesis of endothelial cells (ECs). Confocal analyses of several matrix proteins reveal high intra- and extra-cellular deposition of fibronectin in formed vascular structures. Using a small peptide inhibitor of fibronectin polymerization, we demonstrate that inhibition of fibronectin fibrillogenesis in ECs cultured atop de-cellularized ECM resulted in decreased vascular morphogenesis. Further, immunofluorescence and ultrastructural analyses reveal decreased expression of stromal matrix proteins in the absence of polymerized fibronectin with high co-localization of matrix proteins found in association with polymerized fibronectin. Evaluating vascular kinetics, live cell imaging showed that migration, migration velocity, and mean square displacement, are disrupted in structures grown in the absence of polymerized fibronectin. Additionally, vascular organization failed to occur in the absence of a polymerized fibronectin matrix. Consistent with these observations, we tested vascular morphogenesis following the disruption of EC adhesion to polymerized fibronectin, demonstrating that block of integrins α5β1 and αvβ3, abrogated vascular morphogenesis. Overall, fibronectin deposition in a 3D cell-derived de-cellularized ECM appears to be imperative for matrix assembly and vascular morphogenesis.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, 21218, United States of America.|Johns Hopkins Physical Sciences-Oncology Center, Johns Hopkins University, Baltimore, Maryland, 21218, United States of America.|Department of Biomedical Sciences, Georgia Philadelphia College of Osteopathic Medicine, Suwanee, Georgia, 30024, United States of America.
authors.forename: Abigail
authors.initials: A
authors.lastname: Hielscher
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, 21218, United States of America.
authors.forename: Kim
authors.initials: K
authors.lastname: Ellis
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, 21218, United States of America.
authors.forename: Connie
authors.initials: C
authors.lastname: Qiu
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, 21218, United States of America.
authors.forename: Josh
authors.initials: J
authors.lastname: Porterfield
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, 21218, United States of America.|Johns Hopkins Physical Sciences-Oncology Center, Johns Hopkins University, Baltimore, Maryland, 21218, United States of America.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, Maryland, 21218, United States of America.
authors.forename: Sharon
authors.initials: S
authors.lastname: Gerecht
mesh_terms: D000906:Antibodies / Q000276:immunology; D002460:Cell Line; D002465:Cell Movement; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000276:immunology / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D060888:Real-Time Polymerase Chain Reaction; D019030:Receptors, Vitronectin / Q000276:immunology / Q000378:metabolism; D059008:Time-Lapse Imaging
chemical_list: D000906:Antibodies; D005353:Fibronectins; D039302:Integrin alphaVbeta3; D019030:Receptors, Vitronectin; C094340:integrin alphavbeta1; D003094:Collagen; D020782:Matrix Metalloproteinases
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