Effects of Dexamethasone on Mesenchymal Stromal Cell Chondrogenesis and Aggrecanase Activity

Objective: Dexamethasone (Dex) is a synthetic glucocorticoid that has pro-anabolic and anticatabolic effects in cartilage tissue engineering systems, though the mechanisms by which these effects are mediated are not well understood. We tested the hypothesis that the addition of Dex to chondrogenic medium would affect matrix production and aggrecanase activity of human and bovine bone marrow stromal cells (BMSCs) cultured in self-assembling peptide and agarose hydrogels. Design: We cultured young bovine and adult human BMSCs in (RADA)4 self-assembling peptide and agarose hydrogels in medium containing TGF-β1±Dex and analyzed extracellular matrix composition, aggrecan cleavage products, and the effects of the glucocorticoid receptor antagonist RU-486 on proteoglycan content, synthesis, and catabolic processing. Results: Dex improved proteoglycan synthesis and retention in agarose hydrogels seeded with young bovine cells but decreased proteoglycan accumulation in peptide scaffolds. These effects were mediated by the glucocorticoid receptor. Adult human BMSCs showed minimal matrix accumulation in agarose, but accumulated ~50% as much proteoglycan and collagen as young bovine BMSCs in peptide hydrogels. Dex reduced aggrecanase activity in (RADA)4 and agarose hydrogels, as measured by anti-NITEGE Western blotting, for both bovine and human BMSC-seeded gels. Conclusions: The effects of Dex on matrix production are dependent on cell source and hydrogel identity. This is the first report of Dex reducing aggrecanase activity in a tissue engineering culture system.

[1]  E. Hunziker,et al.  Differential effects of dexamethasone on the chondrogenesis of mesenchymal stromal cells: influence of microenvironment, tissue origin and growth factor. , 2011, European cells & materials.

[2]  A. Grodzinsky,et al.  Effects of short-term glucocorticoid treatment on changes in cartilage matrix degradation and chondrocyte gene expression induced by mechanical injury and inflammatory cytokines , 2011, Arthritis research & therapy.

[3]  A. Grodzinsky,et al.  Growth Factor Delivery Through Self-assembling Peptide Scaffolds , 2011, Clinical orthopaedics and related research.

[4]  I. Erickson,et al.  Cartilage Matrix Formation by Bovine Mesenchymal Stem Cells in Three-dimensional Culture Is Age-dependent , 2011, Clinical orthopaedics and related research.

[5]  M. Endres,et al.  Chondrogenic differentiation of human mesenchymal stem cells in micro‐masses is impaired by high doses of the chemokine CXCL7 , 2011, Journal of tissue engineering and regenerative medicine.

[6]  A. Grodzinsky,et al.  Controlled delivery of transforming growth factor β1 by self-assembling peptide hydrogels induces chondrogenesis of bone marrow stromal cells and modulates Smad2/3 signaling. , 2011, Tissue engineering. Part A.

[7]  A. Augello,et al.  Mesenchymal stem cells from development to postnatal joint homeostasis, aging, and disease. , 2010, Birth defects research. Part C, Embryo today : reviews.

[8]  A. Grodzinsky,et al.  Effect of self-assembling peptide, chondrogenic factors, and bone marrow-derived stromal cells on osteochondral repair. , 2010, Osteoarthritis and cartilage.

[9]  A. Grodzinsky,et al.  Adult equine bone marrow stromal cells produce a cartilage-like ECM mechanically superior to animal-matched adult chondrocytes. , 2010, Matrix biology : journal of the International Society for Matrix Biology.

[10]  A. Grodzinsky,et al.  Self-assembling peptide hydrogels modulate in vitro chondrogenesis of bovine bone marrow stromal cells. , 2010, Tissue engineering. Part A.

[11]  R. Shah,et al.  Supramolecular design of self-assembling nanofibers for cartilage regeneration , 2010, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Xiao-Yu Song,et al.  Mechanical injury potentiates proteoglycan catabolism induced by interleukin-6 with soluble interleukin-6 receptor and tumor necrosis factor alpha in immature bovine and adult human articular cartilage. , 2009, Arthritis and rheumatism.

[13]  M. Levenston,et al.  Chondrocytes and meniscal fibrochondrocytes differentially process aggrecan during de novo extracellular matrix assembly. , 2009, Tissue engineering. Part A.

[14]  C. Pilapil,et al.  Interleukin‐1β and tumor necrosis factor α inhibit chondrogenesis by human mesenchymal stem cells through NF‐κB–dependent pathways , 2009 .

[15]  C. Pilapil,et al.  Osteogenic potential of reamer irrigator aspirator (RIA) aspirate collected from patients undergoing hip arthroplasty , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[16]  R. Tuan,et al.  Mesenchymal stem cells in arthritic diseases , 2008, Arthritis research & therapy.

[17]  C. Archer,et al.  Cartilage integration: evaluation of the reasons for failure of integration during cartilage repair. A review. , 2008, European cells & materials.

[18]  R. Tuan,et al.  Technology Insight: adult mesenchymal stem cells for osteoarthritis therapy , 2008, Nature Clinical Practice Rheumatology.

[19]  W. Richter,et al.  Chondrogenesis of mesenchymal stem cells in gel-like biomaterials in vitro and in vivo. , 2008, Frontiers in bioscience : a journal and virtual library.

[20]  A. Grodzinsky,et al.  Structure of pericellular matrix around agarose-embedded chondrocytes. , 2007, Osteoarthritis and cartilage.

[21]  K. Shimizu,et al.  G1-G2 aggrecan product that can be generated by M-calpain on truncation at Ala709-Ala710 is present abundantly in human articular cartilage. , 2007, Journal of biochemistry.

[22]  J. Connelly,et al.  Dynamic Compression Regulates the Expression and Synthesis of Chondrocyte‐Specific Matrix Molecules in Bone Marrow Stromal Cells , 2007, Stem cells.

[23]  R. Tuan,et al.  Glucocorticoids Promote Chondrogenic Differentiation of Adult Human Mesenchymal Stem Cells by Enhancing Expression of Cartilage Extracellular Matrix Genes , 2006, Stem cells.

[24]  Richard T. Lee,et al.  Local myocardial insulin-like growth factor 1 (IGF-1) delivery with biotinylated peptide nanofibers improves cell therapy for myocardial infarction. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[25]  R. Tuan,et al.  Chondrogenic differentiation and functional maturation of bovine mesenchymal stem cells in long-term agarose culture. , 2006, Osteoarthritis and cartilage.

[26]  R. Tuan,et al.  A three-dimensional nanofibrous scaffold for cartilage tissue engineering using human mesenchymal stem cells. , 2005, Biomaterials.

[27]  P. Neame,et al.  The link proteins , 1993, Experientia.

[28]  P. Chockalingam,et al.  Release of hyaluronan and hyaladherins (aggrecan G1 domain and link proteins) from articular cartilage exposed to ADAMTS-4 (aggrecanase 1) or ADAMTS-5 (aggrecanase 2). , 2004, Arthritis and rheumatism.

[29]  H. Cheung,et al.  Chondrogenesis of human bone marrow-derived mesenchymal stem cells in agarose culture. , 2004, The anatomical record. Part A, Discoveries in molecular, cellular, and evolutionary biology.

[30]  F. Barry,et al.  Mesenchymal stem cells: clinical applications and biological characterization. , 2004, The international journal of biochemistry & cell biology.

[31]  P. Patwari,et al.  Influence of tissue maturation and antioxidants on the apoptotic response of articular cartilage after injurious compression. , 2004, Arthritis and rheumatism.

[32]  Robert Schweitzer,et al.  Glucocorticoid receptor antagonism by cyproterone acetate and RU486. , 2003, Molecular pharmacology.

[33]  F. Barry,et al.  Chondrogenic differentiation of human mesenchymal stem cells within an alginate layer culture system , 2002, In Vitro Cellular & Developmental Biology - Animal.

[34]  A. J. Grodzinsky,et al.  Self-assembling peptide hydrogel fosters chondrocyte extracellular matrix production and cell division: Implications for cartilage tissue repair , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[35]  M. Tortorella,et al.  Inhibition of ADAM-TS4 and ADAM-TS5 Prevents Aggrecan Degradation in Osteoarthritic Cartilage* , 2002, The Journal of Biological Chemistry.

[36]  W. Geissler,et al.  Allosteric Effects of Dexamethasone and RU486 on Glucocorticoid Receptor-DNA Interactions* , 2002, The Journal of Biological Chemistry.

[37]  B Kurz,et al.  Redifferentiation of dedifferentiated bovine articular chondrocytes in alginate culture under low oxygen tension. , 2002, Osteoarthritis and cartilage.

[38]  J. Sandy,et al.  Analysis of aggrecan in human knee cartilage and synovial fluid indicates that aggrecanase (ADAMTS) activity is responsible for the catabolic turnover and loss of whole aggrecan whereas other protease activity is required for C-terminal processing in vivo. , 2001, The Biochemical journal.

[39]  F. Barry,et al.  Gelatin-based resorbable sponge as a carrier matrix for human mesenchymal stem cells in cartilage regeneration therapy. , 2000, Journal of biomedical materials research.

[40]  J. Sandy,et al.  The intermediates of aggrecanase-dependent cleavage of aggrecan in rat chondrosarcoma cells treated with interleukin-1. , 2000, The Biochemical journal.

[41]  R. Tuan,et al.  Cellular interactions and signaling in cartilage development. , 2000, Osteoarthritis and cartilage.

[42]  R Scapinelli,et al.  Hyaluronan-based biopolymers as delivery vehicles for bone-marrow-derived mesenchymal progenitors. , 2000, Journal of biomedical materials research.

[43]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[44]  V. Goldberg,et al.  The Chondrogenic Potential of Human Bone-Marrow-Derived Mesenchymal Progenitor Cells* , 1998, The Journal of bone and joint surgery. American volume.

[45]  A I Caplan,et al.  In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. , 1998, Experimental cell research.

[46]  A. Grodzinsky,et al.  Compression of cartilage results in differential effects on biosynthetic pathways for aggrecan, link protein, and hyaluronan. , 1996, Archives of biochemistry and biophysics.

[47]  A. Grodzinsky,et al.  Fluorometric assay of DNA in cartilage explants using Hoechst 33258. , 1988, Analytical biochemistry.

[48]  R W Farndale,et al.  A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures. , 1982, Connective tissue research.

[49]  H. Stegemann,et al.  Determination of hydroxyproline. , 1967, Clinica chimica acta; international journal of clinical chemistry.