title: Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions.
abstract: Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility.
authors.affiliation: Department of Bioengineering, University of California San Diego, La Jolla, California 92093, USA.|Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, Baltimore, Maryland 21218, USA.|Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Stephanie I
authors.initials: SI
authors.lastname: Fraley
authors.affiliation: Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, Baltimore, Maryland 21218, USA.|Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Pei-Hsun
authors.initials: PH
authors.lastname: Wu
authors.affiliation: Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, Baltimore, Maryland 21218, USA.|Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Lijuan
authors.initials: L
authors.lastname: He
authors.affiliation: Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins University, Baltimore, Maryland 21218, USA.|Departments of Medicine and Cell Biology and Physiology and BRIGHT Institute, Washington University School of Medicine, St. Louis, Missouri 63110, USA.|Department of Pathology, Geisel School of Medicine at Dartmouth, Lebanon, New Hampshire 03756, USA.
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Ranjini
authors.initials: R
authors.lastname: Krisnamurthy
authors.affiliation: Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins University, Baltimore, Maryland 21218, USA.|Departments of Medicine and Cell Biology and Physiology and BRIGHT Institute, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Longmore
authors.affiliation: Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, Baltimore, Maryland 21218, USA.|Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Denis
authors.initials: D
authors.lastname: Wirtz
mesh_terms: D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002465:Cell Movement; D048430:Cell Shape; D003470:Culture Media; D005109:Extracellular Matrix / Q000201:enzymology; D024022:Fibrillar Collagens / Q000737:chemistry / Q000648:ultrastructure; D019204:GTP-Binding Proteins; D006801:Humans; D053511:Matrix Metalloproteinase 14 / Q000502:physiology*; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D020836:Protein Structure, Quaternary; D011503:Transglutaminases
chemical_list: D003470:Culture Media; D024022:Fibrillar Collagens; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14; D019204:GTP-Binding Proteins