Restricted accessResearch articleFirst published online 2010-10
Modulation of Matrix Metalloprotease-2 Levels by Mechanical Loading of Three-Dimensional Mesenchymal Stem Cell Constructs: Impact on In Vitro Tube Formation
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 Hz, 72 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.
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References
1.
GriffithL.G., NaughtonG.Tissue engineering—current challenges and expanding opportunities. Science, 295:1009. 2002.
2.
AdamsR.H., AlitaloK.Molecular regulation of angiogenesis and lymphangiogenesis. Nat Rev Mol Cell Biol, 8:464. 2007.
Von Offenberg SweeneyN., CumminsP.M., CotterE.J., FitzpatrickP.A., BirneyY.A., RedmondE.M., CahillP.A.Cyclic strain-mediated regulation of vascular endothelial cell migration and tube formation. Biochem Biophys Res Commun, 329:573. 2005.
5.
MorrowD., CullenJ.P., CahillP.A., RedmondE.M.Cyclic strain regulates the Notch/CBF-1 signaling pathway in endothelial cells: role in angiogenic activity. Arterioscler Thromb Vasc Biol, 27:1289. 2007.
6.
ShuklaA., DunnA.R., MosesM.A., Van VlietK.J.Endothelial cells as mechanical transducers: enzymatic activity and network formation under cyclic strain. Mech Chem Biosyst, 1:279. 2004.
7.
IchiokaS., ShibataM., KosakiK., SatoY., HariiK., KamiyaA.Effects of shear stress on wound-healing angiogenesis in the rabbit ear chamber. J Surg Res, 72:29. 1997.
8.
SarmientoA., LattaL.L., TarrR.R.The effects of function in fracture healing and stability. Instr Course Lect, 33:83. 1984.
9.
BrownM.D., HudlickaO.Modulation of physiological angiogenesis in skeletal muscle by mechanical forces: involvement of VEGF and metalloproteinases. Angiogenesis, 6:1. 2003.
10.
PietramaggioriG., LiuP., SchererS.S., KaipainenA., PrsaM.J., MayerH., NewalderJ., AlperovichM., MentzerS.J., KonerdingM.A., HuangS., IngberD.E., OrgillD.P.Tensile forces stimulate vascular remodeling and epidermal cell proliferation in living skin. Ann Surg, 246:896. 2007.
GhajarC.M., BlevinsK.S., HughesC.C., GeorgeS.C., PutnamA.J.Mesenchymal stem cells enhance angiogenesis in mechanically viable prevascularized tissues via early matrix metalloproteinase upregulation. Tissue Eng, 12:2875. 2006.
13.
MoioliE.K., ClarkP.A., ChenM., DennisJ.E., EricksonH.P., GersonS.L., MaoJ.J.Synergistic actions of hematopoietic and mesenchymal stem/progenitor cells in vascularizing bioengineered tissues. PLoS One, 3:e3922. 2008.
14.
GruberR., KandlerB., HolzmannP., Vogele-KadletzM., LosertU., FischerM.B., WatzekG.Bone marrow stromal cells can provide a local environment that favors migration and formation of tubular structures of endothelial cells. Tissue Eng, 11:896. 2005.
15.
MarkowiczM., KoellenspergerE., NeussS., SteffensG.C.M., PalluaN.Enhancing the vascularization of three-dimensional scaffolds: new strategies in tissue regeneration and tissue engineering. Top Tissue Eng, 2:1. 2005.
16.
KasperG., DankertN., TuischerJ., HoeftM., GaberT., GlaeserJ.D., ZanderD., TschirschmannM., ThompsonM., MatziolisG., DudaG.N.Mesenchymal stem cells regulate angiogenesis according to their mechanical environment. Stem Cells, 25:903. 2007.
17.
Page-McCawA., EwaldA.J., WerbZ.Matrix metalloproteinases and the regulation of tissue remodelling. Nat Rev Mol Cell Biol, 8:221. 2007.
18.
SternlichtM.D., WerbZ.How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol, 17:463. 2001.
19.
KasperG., GlaeserJ.D., GeisslerS., OdeA., TuischerJ., MatziolisG., PerkaC., DudaG.N.Matrix metalloprotease activity is an essential link between mechanical stimulus and mesenchymal stem cell behavior. Stem Cells, 25:1985. 2007.
20.
RiesC., EgeaV., KarowM., KolbH., JochumM., NethP.MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines. Blood, 109:4055. 2007.
21.
SeliktarD., NeremR.M., GalisZ.S.The role of matrix metalloproteinase-2 in the remodeling of cell-seeded vascular constructs subjected to cyclic strain. Ann Biomed Eng, 29:923. 2001.
22.
ButlerG.S., ButlerM.J., AtkinsonS.J., WillH., TamuraT., Schade van WestrumS., CrabbeT., ClementsJ., d'OrthoM.P., MurphyG.The TIMP2 membrane type 1 metalloproteinase “receptor” regulates the concentration and efficient activation of progelatinase A. A kinetic study. J Biol Chem, 273:871. 1998.
23.
WangZ., JuttermannR., SolowayP.D.TIMP-2 is required for efficient activation of proMMP-2 in vivo. J Biol Chem, 275:26411. 2000.
24.
StronginA.Y., CollierI., BannikovG., MarmerB.L., GrantG.A., GoldbergG.I.Mechanism of cell surface activation of 72-kDa type IV collagenase. Isolation of the activated form of the membrane metalloprotease. J Biol Chem, 270:5331. 1995.
25.
SounniN.E., JanssenM., FoidartJ.M., NoelA.Membrane type-1 matrix metalloproteinase and TIMP-2 in tumor angiogenesis. Matrix Biol, 22:55. 2003.
26.
YanaI., WeissS.J.Regulation of membrane type-1 matrix metalloproteinase activation by proprotein convertases. Mol Biol Cell, 11:2387. 2000.
27.
DominiciM., Le BlancK., MuellerI., Slaper-CortenbachI., MariniF., KrauseD., DeansR., KeatingA., ProckopD., HorwitzE.Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315. 2006.
28.
PittengerM.F., MackayA.M., BeckS.C., JaiswalR.K., DouglasR., MoscaJ.D., MoormanM.A., SimonettiD.W., CraigS., MarshakD.R.Multilineage potential of adult human mesenchymal stem cells. Science, 284:143. 1999.
29.
AdesE.W., CandalF.J., SwerlickR.A., GeorgeV.G., SummersS., BosseD.C., LawleyT.J.HMEC-1: establishment of an immortalized human microvascular endothelial cell line. J Invest Dermatol, 99:683. 1992.
30.
MatziolisG., TuischerJ., KasperG., ThompsonM., BartmeyerB., KrockerD., PerkaC., DudaG.Simulation of cell differentiation in fracture healing: mechanically loaded composite scaffolds in a novel bioreactor system. Tissue Eng, 12:201. 2006.
31.
DuongH., WuB., TawilB.Modulation of 3D fibrin matrix stiffness by intrinsic fibrinogen-thrombin compositions and by extrinsic cellular activity. Tissue Eng Part A, 15:1865. 2009.
32.
CholewinskiE., DietrichM., FlanaganT.C., Schmitz-RodeT., JockenhoevelS.Tranexamic acid—an alternative to aprotinin in fibrin-based cardiovascular tissue engineering. Tissue Eng Part A, 15:3645. 2009.
33.
BaumgartnerL., ArnholdS., BrixiusK., AddicksK., BlochW.Human mesenchymal stem cells: influence of oxygen pressure on proliferation and chondrogenic differentiation in fibrin glue in vitro. J Biomed Mater Res A, 93:930. 2010.
34.
CatelasI., SeseN., WuB.M., DunnJ.C., HelgersonS., TawilB.Human mesenchymal stem cell proliferation and osteogenic differentiation in fibrin gels in vitro. Tissue Eng, 12:2385. 2006.
35.
FujiwaraT., OdaK., YokotaS., TakatsukiA., IkeharaY.Brefeldin A causes disassembly of the Golgi complex and accumulation of secretory proteins in the endoplasmic reticulum. J Biol Chem, 263:18545. 1988.
36.
LyonsA.B., ParishC.R.Determination of lymphocyte division by flow cytometry. J Immunol Methods, 171:131. 1994.
37.
BergerT.G., FeuersteinB., StrasserE., HirschU., SchreinerD., SchulerG., Schuler-ThurnerB.Large-scale generation of mature monocyte-derived dendritic cells for clinical application in cell factories. J Immunol Methods, 268:131. 2002.
38.
GoodshipA.E., KenwrightJ.The influence of induced micromovement upon the healing of experimental tibial fractures. J Bone Joint Surg Br, 67:650. 1985.
39.
IvanenkoY.P., GrassoR., LacquanitiF.Influence of leg muscle vibration on human walking. J Neurophysiol, 84:1737. 2000.
40.
TurnerC.H., ForwoodM.R., OtterM.W.Mechanotransduction in bone: do bone cells act as sensors of fluid flow?FASEB J, 8:875. 1994.
41.
BrownL.F., LanirN., McDonaghJ., TognazziK., DvorakA.M., DvorakH.F.Fibroblast migration in fibrin gel matrices. Am J Pathol, 142:273. 1993.
42.
SehgalI., ThompsonT.C.Novel regulation of type IV collagenase (matrix metalloproteinase-9 and -2) activities by transforming growth factor-beta1 in human prostate cancer cell lines. Mol Biol Cell, 10:407. 1999.
GuoC., PiacentiniL.Type I collagen-induced MMP-2 activation coincides with up-regulation of membrane type 1-matrix metalloproteinase and TIMP-2 in cardiac fibroblasts. J Biol Chem, 278:46699. 2003.
45.
MottJ.D., WerbZ.Regulation of matrix biology by matrix metalloproteinases. Curr Opin Cell Biol, 16:558. 2004.
46.
BelottiD., PaganoniP., ManentiL., GarofaloA., MarchiniS., TarabolettiG., GiavazziR.Matrix metalloproteinases (MMP9 and MMP2) induce the release of vascular endothelial growth factor (VEGF) by ovarian carcinoma cells: implications for ascites formation. Cancer Res, 63:5224. 2003.
47.
XuJ., RodriguezD., PetitclercE., KimJ.J., HangaiM., MoonY.S., DavisG.E., BrooksP.C.Proteolytic exposure of a cryptic site within collagen type IV is required for angiogenesis and tumor growth in vivo. J Cell Biol, 154:1069. 2001.
48.
EnensteinJ., WalehN.S., KramerR.H.Basic FGF and TGF-beta differentially modulate integrin expression of human microvascular endothelial cells. Exp Cell Res, 203:499. 1992.
49.
ChakrabortiS., MandalM., DasS., MandalA., ChakrabortiT.Regulation of matrix metalloproteinases: an overview. Mol Cell Biochem, 253:269. 2003.
50.
LiA., DubeyS., VarneyM.L., DaveB.J., SinghR.K.IL-8 directly enhanced endothelial cell survival, proliferation, and matrix metalloproteinases production and regulated angiogenesis. J Immunol, 170:3369. 2003.
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