title: Modulation of matrix metalloprotease-2 levels by mechanical loading of three-dimensional mesenchymal stem cell constructs: impact on in vitro tube formation.
abstract: Angiogenesis is essential to tissue reconstitution, is sensitive to mechanical stresses, and currently represents one of the major challenges in tissue engineering. The pro-angiogenic matrix metalloprotease-2 (MMP-2) is upregulated in mechanically loaded mesenchymal stem cells (MSCs). Therefore, MMP-2 may provide a regulating link between angiogenesis and the surrounding mechanical conditions. This study aimed to modulate MMP-2 levels by mechanical loading of MSCs embedded in a three-dimensional matrix as well as to investigate the mechanism of MMP-2 regulation along with its contribution to angiogenesis stimulation. MMP-2-inducing conditions (30% compression, 1&#x2009;Hz, 72&#x2009;h) were defined after varying loading parameters. Addition of the Golgi-disturbing agent Brefeldin A suppressed this mechanical upregulation of MMP-2. Analysis of enzymatic activities demonstrated an enhancement of pro-MMP-2, mature MMP-2, and tissue inhibitor of metalloproteases-2. Further, mechano-regulation of MMP-14 and mature MMP-2 was dependent upon the activity of furin, a proprotein processing endoprotease. Angiogenesis was stimulated by conditioned media from MSCs loaded at inducing conditions. This augmentation of angiogenesis was hindered by inhibition of pro-MMP-2 and mature MMP-2. In conclusion, mechanical stimulation of MSCs in a three-dimensional matrix induces pro-MMP-2 secretion and MMP-2 activation, potentially via the activation complex consisting of MMP-2/-14/tissue inhibitor of metalloproteases-2. Mechano-regulated pro-MMP-2 and mature MMP-2 seem to contribute to angiogenesis stimulation. Thus, an application of these loading parameters could augment vascularization of tissue-engineered constructs based on the described MMP-2-dependent mechanism.
authors.affiliation: Julius Wolff Institute and Center for Musculoskeletal Surgery, Charité—Universitätsmedizin , Berlin, Germany.
authors.forename: Juliane D
authors.initials: JD
authors.lastname: Glaeser
authors.forename: Sven
authors.initials: S
authors.lastname: Geissler
authors.forename: Andrea
authors.initials: A
authors.lastname: Ode
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Schipp
authors.forename: Georg
authors.initials: G
authors.lastname: Matziolis
authors.forename: William R
authors.initials: WR
authors.lastname: Taylor
authors.forename: Petra
authors.initials: P
authors.lastname: Knaus
authors.forename: Carsten
authors.initials: C
authors.lastname: Perka
authors.forename: Georg N
authors.initials: GN
authors.lastname: Duda
authors.forename: Grit
authors.initials: G
authors.lastname: Kasper
mesh_terms: D001696:Biomechanical Phenomena / Q000502:physiology*; D019149:Bioreactors; D049109:Cell Proliferation; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D018919:Neovascularization, Physiologic / Q000502:physiology*; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D020778:Matrix Metalloproteinase 2