Mechanical compression of cartilage explants induces multiple time-dependent gene expression patterns and involves intracellular calcium and cyclic AMP

Fitzgerald, Jonathan B.<sup>1</sup>; Jin, Moonsoo<sup>2,5</sup>; Dean, Delphine<sup>3</sup>; Wood, David J.<sup>4</sup>; Zheng, Ming H.<sup>4</sup>; Grodzinsky, Alan J.<sup>1,2,3</sup>

Affiliations

¹Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

²Center for Biomedical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

³Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

⁴Department of Orthopaedic Surgery, QEII Medical Centre, University of Western Australia, Nedlands, Western Australia 6009, Australia

⁵Present address: The CBR Institute for Biomedical Research, Inc., Harvard Medical School, Boston, MA 02115

Abstract

Chondrocytes are influenced by mechanical forces to remodel cartilage extracellular matrix. Previous studies have demonstrated the effects of mechanical forces on changes in biosynthesis and mRNA levels of particular extracellular matrix molecules, and have identified certain signaling pathways that may be involved. However, the broad extent and kinetics of mechano-regulation of gene transcription has not been studied in depth. We applied static compressive strains to bovine cartilage explants for periods between 1 and 24 h and measured the response of 28 genes using real time PCR. Compression time courses were also performed in the presence of an intracellular calcium chelator or an inhibitor of cyclic AMP-activated protein kinase A. Cluster analysis of the data revealed four main expression patterns: two groups containing either transiently up-regulated or duration-enhanced expression profiles could each be subdivided into genes that did or did not require intracellular calcium release and cyclic AMP-activated protein kinase A for their mechano-regulation. Transcription levels for aggrecan, type II collagen, and link protein were up-regulated ∼2–3-fold during the first 8 h of 50% compression and subsequently down-regulated to levels below that of free-swelling controls by 24 h. Transcription levels of matrix metalloproteinases-3, -9, and -13, aggrecanase-1, and the matrix protease regulator cyclooxygenase-2 increased with the duration of 50% compression 2–16-fold by 24 h. Thus, transcription of proteins involved in matrix remodeling and catabolism dominated over anabolic matrix proteins as the duration of static compression increased. Immediate early genes c-fos and c-jun were dramatically up-regulated 6–30-fold, respectively, during the first 8 h of 50% compression and remained up-regulated after 24 h.

Links

DOI: 10.1074/jbc.M400437200

PubMed: 14960571

WoS: 000221164500017

History

Received: 2004-01-14

Revised: 2004-05-7

Cited Works (16)

Year	Entry
1999	Herberhold C, Faber S, Stammberger T, Steinlechner M, Putz R, Englmeier K, Reiser M, Eckstein F. In situ measurement of articular cartilage deformation in intact femoropatellar joints under static loading. J Biomech. December 1999;32(12):1287-1295.
2002	Davisson T, Kunig S, Chen A, Sah R, Ratcliffe A. Static and dynamic compression modulate matrix metabolism in tissue engineered cartilage. J Orthop Res. July 2002;20(4):842-848.
2000	Grodzinsky AJ, Levenston ME, Jin M, Frank EH. Cartilage tissue remodeling in response to mechanical forces. Annu Rev Biomed Eng. 2000;2:691-713.
1994	Guilak F, Meyer BC, Ratcliffe A, Mow VC. The effects of matrix compression on proteoglycan metabolism in articular cartilage explants. Osteoarthritis Cartilage. June 1994;2(2):91-101.
1994	Kim Y-J, Sah RLY, Grodzinsky AJ, Plaas AHK, Sandy JD. Mechanical regulation of cartilage biosynthetic behavior: physical stimuli. Arch Biochem Biophys. May 1994;311(1):1-12.
2002	Thibault M, Poole AR, Buschmann MD. Cyclic compression of cartilage/bone explants in vitro leads to physical weakening, mechanical breakdown of collagen and release of matrix fragments. J Orthop Res. November 2002;20(6):1265-1273.
2001	Bonassar LJ, Grodzinsky AJ, Frank EH, Davila SG, Bhaktav NR, Trippel SB. The effect of dynamic compression on the response of articular cartilage to insulin-like growth factor-i. J Orthop Res. January 2001;19(1):11-17.
2000	Bonassar LJ, Grodzinsky AJ, Srinivasan A, Davila SG, Trippel SB. Mechanical and physicochemical regulation of the action of insulin-like growth factor-I on articular cartilage. Arch Biochem Biophys. July 1, 2000;379(1):57-63.
2000	Mauck RL, Soltz MA, Wang CCB, Wong DD, Chao P-HG, Valhmu WB, Hung CT, Ateshian GA. Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels. J Biomech Eng. June 2000;122(3):252-260.
2001	Jin M, Frank EH, Quinn TM, Hunziker EB, Grodzinsky AJ. Tissue shear deformation stimulates proteoglycan and protein biosynthesis in bovine cartilage explants. Arch Biochem Biophys. November 1, 2001;395(1):41-48.
1986	Hodge WA, Fijan RS, Carlson KL, Burgess RG, Harris WH, Mann RW. Contact pressures in the human hip joint measured in vivo. Proc Natl Acad Sci USA. May 1986;83(9):2879-2883.
1999	Ragan PM, Badger AM, Cook M, Chin VI, Gowen M, Grodzinsky AJ, Lark MW. Down-regulation of chondrocyte aggrecan and type-II collagen gene expression correlates with increases in static compression magnitude and duration. J Orthop Res. November 1999;17(6):836-842.
2003	Ikenoue T, Trindade MCD, Lee MS, Lin EY, Schurman DJ, Goodman SB, Smith RL. Mechanoregulation of human articular chondrocyte aggrecan and type II collagen expression by intermittent hydrostatic pressure in vitro. J Orthop Res. January 2003;21(1):110-116.
1998	Valhmu WB, Stazzone EJ, Bachrach NM, Saed-Nejad F, Fischer SG, Mow VC, Ratcliffe A. Load-controlled compression of articular cartilage induces a transient stimulation of aggrecan gene expression. Arch Biochem Biophys. May 1, 1998;353(1):29-36.
2004	Kisiday JD, Jin M, DiMicco MA, Kurz B, Grodzinsky AJ. Effects of dynamic compressive loading on chondrocyte biosynthesis in self-assembling peptide scaffolds. J Biomech. May 2004;37(5):595-604.
2002	Hunter CJ, Imler SM, Malaviya P, Nerem RM, Levenston ME. Mechanical compression alters gene expression and extracellular matrix synthesis by chondrocytes cultured in collagen I gels. Biomaterials. February 15, 2002;23(4):1249-1259.

Cited By (33)

Year	Entry
2007	Mauck RL, Byers BA, Yuan X, Tuan RS. Regulation of cartilaginous ECM gene transcription by chondrocytes and MSCs in 3D culture in response to dynamic loading. Biomech Model Mechanobiol. January 2007;6(1-2):113-125.
2014	Sanchez-Adams J, Leddy HA, McNulty AL, O’Conor CJ, Guilak F. The mechanobiology of articular cartilage: bearing the burden of osteoarthritis. Curr Rheumatol Rep. October 2014;16(10):451.
2019	Franklin ME. Structural Characterization and Introduction of Biomimetic Proteoglycans in Temporomandibular Joint Pain [Master's thesis]. Drexel University; June 2019.
2016	Schätti OR. A Model to Study Articular Cartilage Mechanical and Biological Responses to Sliding and Rolling Loads [PhD thesis]. Swiss Federal Institute of Technology Zürich; 2016.
2006	Gemmiti CV. Effect of Fluid Flow on Tissue-Engineered Cartilage in a Novel Bioreactor [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; December 2006.
2007	Connelly JT. Regulatory Mechanisms in the Chondrogenesis of Mesenchymal Progenitors: The Roles of Cyclic Tensile Loading and Cell-Matrix Interactions [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; August 2007.
2005	FitzGerald JB. Chondrocyte Gene Expression and Intracellular Signaling Pathways in Cartilage Mechanotransduction [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; July 2005.
2005	Lee JH. Chondrocyte Response to in Vitro Mechanical Injury and Co-Culture With Joint Capsule Tissue [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; June 2005.
2005	Ng LJ. Probing Nanomechanics of Aggrecan and the Aggrecan-Rich Pericellular Matrix of Chondrocytes in Cartilage [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; September 2005.
2005	Szafranski JD. Cartilage Mechanobiology: The Effects of Loading on the Fine Structure and Function of Chondroitin Sulfate Glycosaminoglycans [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; June 2005.
2008	Chai DH. Progression of Chondrocyte Signaling Responses to Mechanical Stimulation in 3-D Gel Culture [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; February 2008.
2008	Wheeler CA. Cartilage Response to in Vitro Models of Injury in Combination With Growth Factor and Antioxidant Treatments [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; February 2008.
2009	Chen S. Regulation of Lubricin Gene Expression and Synthesis in Cartilage by Mechanical Injury [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; September 2009.
2009	Kopesky PW. Self-Assembling Peptide Hydrogels Promote in Vitro Chondrogenesis of Bone Marrow-Derived Stromal Cells: Effects of Peptide Sequence, Cell Donor Age, and Method of Growth Factor Delivery [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; June 2009.
2010	Lu YCS. In Vitro Models of Cartilage Degradation Following Joint Injury: Mechanical Overload, Inflammatory Cytokines and Therapeutic Approaches [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; February 2010.
2013	Li Y. In Vitro Model of Injury/cytokine-Induced Cartilage Catabolism Modulated by Dynamic Compression, Growth Factors, and Glucocorticoids [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; June 2013.
2016	Wang Y. Molecular Pathway Analysis and Therapeutics Development in Post-Traumatic Osteoarthritis [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; September 2016.
2018	Salinas Duron D. New Methods in Computation of Reaction Fluxes from Metabolomics Data [PhD thesis]. Bozeman, MT: Montana State University; April 2018.
2010	Killian ML. The Influence of Traumatic Impaction and Pathological Loading on Knee Menisci [PhD thesis]. Houghton, MI: Michigan Technological University; 2010.
2012	Kaupp JA. Simulated Joint Loading Enhances the Expression of of Superficial Zone Markers in Tissue Engineered Cartilage Constructs [PhD thesis]. Queen's University; February 2012.
2011	Jeon JE. Development of Zonal Cartilage Constructs: Effects of Chondrocyte Subpopulation, Compressive Stimulation, and Culture Models [PhD thesis]. Queensland University of Technology; 2011.
2015	Kuaha K. Environments for Zonal Cartilage Tissue Engineering [PhD thesis]. Queensland University of Technology; 2015.
2013	Zheng Ch. Effects of Biochemical Factors and Mechanical Loading on Chondrogenesis of Human Adipose-Derived Stem Cells [PhD thesis]. Stanford University; December 2013.
2010	Holguin N. Low-Level Vibrations Maintain the Intervertebral Disc During Unloading [PhD thesis]. Stony Brook, NY: Stony Brook University; December 2010.
2013	Madden RMJ. In Situ Chondrocyte Mechanics and Mechanobiology [Master's thesis]. Calgary, AB: University of Calgary; August 2013.
2012	Nguyen QT. Mechanobiology and Biomechanics of Articular Cartilage Repair [PhD thesis]. San Diego (UCSD), University of California; 2012.
2018	Graham B. Novel Approaches to Determine the Role of Diffusive and Convective Transport Environments in Articular Cartilage Function [PhD thesis]. University of Delaware; 2018.
2018	Zhou Y. Intracellular Calcium Signaling of Chondrocytes and the Treatment of Osteoarthritis [PhD thesis]. University of Delaware; 2018.
2021	McDonough RC. Controlled Calcium Activation Via Chemogenetic Signaling Platforms for Tissue Engineering of Articular Cartilage [PhD thesis]. University of Delaware; 2021.
2014	O’Conor CJ. TRPV4-Mediated Mechanotransduction in Articular Cartilage Function and Disease [PhD thesis]. University of North Carolina at Chapel Hill; 2014.
2015	Han WJ. Mechanical Strain Transfer and Calcium Signaling in Fiber-Reinforced Musculoskeletal Soft Tissues [PhD thesis]. Philadelphia, PA: University of Pennsylvania; 2015.
2008	Korecki CL. Effects of Compression Loading, Injury, and Age on Intervertebral Disc Mechanics, Biology and Metabolism Using Large Animal Organ and Cell Culture Systems [PhD thesis]. University of Vermont; May 2008.
2014	Pourmohammadali H. Mechanical and Hydromechanical Stimulation of Chondrocytes for Articular Cartilage Tissue Engineering [PhD thesis]. University of Waterloo; 2014.