Recapitulating aspects of the oxygen and substrate environment of the damaged joint milieu for stem cell-based cartilage tissue engineering

Tissue Engineering - Part C: Methods

Volumn 19, Issue 2, 2013, Pages 117-127

  O'Heireamhoin, Sven ^a   Buckley, Conor T ^a   Jones, Elena ^b   McGonagle, Dennis ^b   Mulhall, Kevin J ^a,c   Kelly, Daniel J ^a  

^a TRINITY COLLEGE DUBLIN   (Ireland)

^b UNIVERSITY OF LEEDS   (United Kingdom)

^c Sports Surgery Clinic   (Ireland)

Author keywords

[No Author keywords available]

Indexed keywords

BIOACTIVITY; CARTILAGE; CELL CULTURE; CELL PROLIFERATION; EXPANSION; HYDROGELS; JOINTS (ANATOMY); OXYGEN; STEM CELLS; TISSUE ENGINEERING;

AGAROSE; ARTICULAR CARTILAGES; BIOLOGICAL FEATURES; CARTILAGE REPAIR; CARTILAGE TISSUE ENGINEERING; CELL POPULATIONS; CELL YIELDS; CELL-BASED; CHONDROGENIC; CHONDROGENIC POTENTIAL; COLONY SIZE; FIBRIN CLOTS; FIBROBLAST GROWTH FACTOR-2; GROWTH FACTOR; HYDROGEL SCAFFOLDS; IN-VITRO; MESENCHYMAL STEM CELL; MICRO-FRACTURE; MICROENVIRONMENTS; MODEL SYSTEM; MONOLAYER EXPANSION; OXYGEN TENSION; POTENTIAL UTILITY; STANDARD EXPANSIONS; SULFATED GLYCOSAMINOGLYCAN (SGAG); TRANSFORMING GROWTH FACTORS;

GROWTH KINETICS;

HARNESS;

FIBROBLAST GROWTH FACTOR 2; OXYGEN;

ARTICLE; CARTILAGE; CELL DIFFERENTIATION; CYTOLOGY; HUMAN; METABOLISM; STEM CELL; TISSUE ENGINEERING;

CARTILAGE; CELL DIFFERENTIATION; FIBROBLAST GROWTH FACTOR 2; HUMANS; OXYGEN; STEM CELLS; TISSUE ENGINEERING;

MLCS; MLOWN;

EID: 84872100990     PISSN: 19373384     EISSN: 19373392     Source Type: Journal    
DOI: 10.1089/ten.tec.2012.0142     Document Type: Article

References (86)

1. Dragoo, J.L., Samimi, B., Zhu, M., Hame, S.L., Thomas, B.J., Lieberman, J.R., et al. Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads. J Bone Joint Surg Br 85, 740, 2003.
2. Wickham, M.Q., Erickson, G.R., Gimble, J.M., Vail, T.P., and Guilak, F. Multipotent stromal cells derived from the infrapatellar fat pad of the knee. Clin Orthop Relat Res 196, 2003.
3. English, A., Jones, E.A., Corscadden, D., Henshaw, K., Chapman, T., Emery, P., et al. A comparative assessment of cartilage and joint fat pad as a potential source of cells for autologous therapy development in knee osteoarthritis. Rheumatology 46, 1676, 2007.
4. Lopa, S., Colombini, A., de Girolamo, L., Sansone, V., and Moretti, M. New Strategies in cartilage tissue engineering for osteoarthritic patients: infrapatellar fat pad as an alternative source of progenitor cells. J Biomater Tissue Eng 1, 40, 2011.
5. Buckley, C.T., Vinardell, T., Thorpe, S.D., Haugh, M.G., Jones, E., McGonagle, D., et al. Functional properties of cartilaginous tissues engineered from infrapatellar fat padderived mesenchymal stem cells. J Biomech 43, 920, 2010.
6. Jurgens, W.J.F.M., Van Dijk, A., Doulabi, B.Z., Niessen, F.B., Ritt, M.J.P.F., Van Milligen, F.J., et al. Freshly isolated stromal cells from the infrapatellar fat pad are suitable for a onestep surgical procedure to regenerate cartilage tissue. Cytotherapy 11, 1052, 2009.
7. Toghraie, F.S., Chenari, N., Gholipour, M.A., Faghih, Z., Torabinejad, S., Dehghani, S., et al. Treatment of osteoarthritis with infrapatellar fat pad derived mesenchymal stem cells in Rabbit. Knee 18, 71, 2011.
8. Marsano, A., Millward-Sadler, S.J., Salter, D.M., Adesida, A., Hardingham, T., Tognana, E., et al. Differential cartilaginous tissue formation by human synovial membrane, fat pad, meniscus cells and articular chondrocytes. Osteoarthritis Cartilage 15, 48, 2007.
9. Vinardell, T., Buckley, C.T., Thorpe, S.D., and Kelly DJ. Composition-function relations of cartilaginous tissues engineered from chondrocytes and mesenchymal stem cells isolated from bone marrow and infrapatellar fat pad. J Tissue Eng Regen Med 5, 673, 2011.
10. Vinardell, T., Sheehy, E.J., Buckley, C.T., and Kelly, D.J. A comparison of the functionality and in vivo phenotypic stability of cartilaginous tissues engineered from different stem cells sources. Tissue Eng Part A 18, 1161, 2012.
11. Lee, S.Y., Nakagawa, T., and Reddi, A.H. Induction of chondrogenesis and expression of superficial zone protein (SZP)/lubricin by mesenchymal progenitors in the infrapatellar fat pad of the knee joint treated with TGF-b1 and BMP-7. Biochem Biophys Res Commun 376, 148, 2008.
12. Lee, S.Y., Nakagawa, T., and Reddi, A.H. Mesenchymal progenitor cells derived from synovium and infrapatellar fat pad as a source for superficial zone cartilage tissue engineering: analysis of superficial zone protein/lubricin expression. Tissue Eng Part A 16, 317, 2010.
13. Solchaga, L.A., Penick, K., Porter, J.D., Goldberg, V.M., Caplan, A.I., and Welter, J.F. FGF-2 enhances the mitotic and chondrogenic potentials of human adult bone marrowderived mesenchymal stem cells. J Cell Physiol 203, 398, 2005.
14. Jung, S., Sen, A., Rosenberg, L., and Behie, L.A. Identification of growth and attachment factors for the serum-free isolation and expansion of human mesenchymal stromal cells. Cytotherapy 12, 637, 2010.
15. Estes, B.T., Diekman, B.O., and Guilak, F. Monolayer cell expansion conditions affect the chondrogenic potential of adipose-derived stem cells. Biotechnol Bioeng 99, 986, 2008.
16. Reyes, M., Lund, T., Lenvik, T., Aguiar, D., Koodie, L., and Verfaillie, C.M. Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98, 2615, 2001.
17. Mastrogiacomo, M., Cancedda, R., and Quarto, R. Effect of different growth factors on the chondrogenic potential of human bone marrow stromal cells. Osteoarthritis Cartilage 9, S36-S40. 2001.
18. Banfi, A., Muraglia, A., Dozin, B., Mastrogiacomo, M., Cancedda, R., and Quarto, R. Proliferation kinetics and differentiation potential of ex vivo expanded human bone marrow stromal cells: Implications for their use in cell therapy. Exp Hematol 28, 707, 2000.
19. Bianchi, G., Banfi, A., Mastrogiacomo, M., Notaro, R., Luzzatto, L., Cancedda, R., et al. Ex vivo enrichment of mesenchymal cell progenitors by fibroblast growth factor 2. Exp Cell Res 287, 98, 2003.
20. Martin, I., Muraglia, A., Campanile, G., Cancedda, R., and Quarto, R. Fibroblast growth factor-2 supports ex vivo expansion and maintenance of osteogenic precursors from human bone marrow. Endocrinology 138, 4456, 1997.
21. Tsutsumi, S., Shimazu, A., Miyazaki, K., Pan, H., Koike, C., Yoshida, E., et al. Retention of multilineage differentiation potential of mesenchymal cells during proliferation in response to FGF. Biochem Biophys Res Commun 288, 413, 2001.
22. Zaragosi, L.E., Ailhaud, G., and Dani, C. Autocrine fibroblast growth factor 2 signaling is critical for self-renewal of human multipotent adipose-derived stem cells. Stem Cells 24, 2412, 2006.
23. Jansen, E.J.P., Emans, P.J., Guldemond, N.A., Van Rhijn, L.W., Welting, T.J.M., Bulstra, S.K., et al. Human periosteumderived cells from elderly patients as a source for cartilage tissue engineering? J Tissue Eng Regen Med 2, 331, 2008.
24. Khan, W.S., Tew, S.R., Adesida, A.B., and Hardingham, T.E. Human infrapatellar fat pad-derived stem cells express the pericyte marker 3G5 and show enhanced chondrogenesis after expansion in fibroblast growth factor-2. Arthritis Res Ther 10, 2008.
25. Buckley, C.T., and Kelly, D.J. Expansion in the presence of FGF-2 enhances the functional development of cartilaginous tissues engineered using infrapatellar fat pad derived MSCs. J Mech Behav Biomed Mater 11, 102, 2012.
26. Solchaga, L.A., Penick, K., Goldberg, V.M., Caplan, A.I., and Welter, J.F. Fibroblast growth factor-2 enhances proliferation and delays loss of chondrogenic potential in human adult bone-marrow-derived mesenchymal stem cells. Tissue Eng Part A 16, 1009, 2010.
27. Estes, B.T., Wu, A.W., Storms, R.W., and Guilak, F. Extended passaging, but not aldehyde dehydrogenase activity, increases the chondrogenic potential of human adipose-derived adult stem cells. J Cell Physiol 209, 987, 2006.
28. Ng, K.W., Lima, E.G., Bian, L., O'Conor, C.J., Jayabalan, P.S., Stoker, A.M., et al. Passaged adult chondrocytes can form engineered cartilage with functional mechanical properties: a canine model. Tissue Eng Part A 16, 1041, 2010.
29. Bian, L., Fong, J.V., Lima, E.G., Stoker, A.M., Ateshian, G.A., Cook, J.L., et al. Dynamic mechanical loading enhances functional properties of tissue-engineered cartilage using mature canine chondrocytes. Tissue Eng Part A 16, 1781, 2010.
30. Jakob, M., Démarteau, O., Schäfer, D., Hintermann, B., Dick, W., Heberer, M., et al. Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro. J Cell Biochem 81, 368, 2001.
31. Khan, I.M., Evans, S.L., Young, R.D., Blain, E.J., Quantock, A.J., Avery, N., et al. Fibroblast growth factor 2 and transforming growth factor b1 induce precocious maturation of articular cartilage. Arthritis Rheum 63, 3417, 2011.
32. Ma, T., Grayson, W.L., Fröhlich, M., and Vunjak-Novakovic, G. Hypoxia and stem cell-based engineering of mesenchymal tissues. Biotechnol Prog 25, 32, 2009.
33. Carrancio, S., López-Holgado, N., Sánchez-Guijo, F.M., Villarón E, Barbado, V., Tabera, S., et al. Optimization of mesenchymal stem cell expansion procedures by cell separation and culture conditions modification. Exp Hematol 36, 1014, 2008.
34. Dos Santos, F., Andrade, P.Z., Boura, J.S., Abecasis, M.M., Da Silva, C.L., and Cabral, J.M.S. Ex vivo expansion of human mesenchymal stem cells: A more effective cell proliferation kinetics and metabolism under hypoxia. J Cell Physiol 223, 27, 2010.
35. Grayson, W.L., Zhao, F., Bunnell, B., and Ma, T. Hypoxia enhances proliferation and tissue formation of human mesenchymal stem cells. Biochem Biophys Res Commun 358, 948, 2007.
36. Grayson, W.L., Zhao, F., Izadpanah, R., Bunnell, B., andMa, T. Effects of hypoxia on human mesenchymal stem cell expansion and plasticity in 3D constructs. J Cell Physiol 207, 331, 2006.
37. Krinner, A., Zscharnack, M., Bader, A., Drasdo, D., and Galle, J. Impact of oxygen environment on mesenchymal stem cell expansion and chondrogenic differentiation. Cell Prolif 42, 471, 2009.
38. Zscharnack, M., Poesel, C., Galle, J., and Bader, A. Low oxygen expansion improves subsequent chondrogenesis of ovine bone-marrow-derived mesenchymal stem cells in collagen type I hydrogel. Cells Tissues Organs 190, 81, 2009.
39. Muller, J., Benz, K., Ahlers, M., Gaissmaier, C., and Mollenhauer, J. Hypoxic conditions during expansion culture prime human mesenchymal stromal precursor cells for chondrogenic differentiation in three-dimensional cultures. Cell Transplant 2011 [Epub ahead of print]; Doi: 10.3727/096368910X564094.
40. Xu, Y., Malladi, P., Chiou, M., Bekerman, E., Giaccia, A.J., and Longaker, M.T. In vitro expansion of adipose-derived adult stromal cells in hypoxia enhances early chondrogenesis. Tissue Eng 13, 2981, 2007.
41. Egli, R.J., Bastian, J.D., Ganz, R., Hofstetter, W., and Leunig, M. Hypoxic expansion promotes the chondrogenic potential of articular chondrocytes. J Orthop Res 26, 977, 2008.
42. Henderson, J.H., Ginley, N.M., Caplan, A.I., Niyibizi, C., and Dennis, J.E. Low oxygen tension during incubation periods of chondrocyte expansion is sufficient to enhance postexpansion chondrogenesis. Tissue Eng Part A 16, 1585, 2010.
43. Wang, D.W., Fermor, B., Gimble, J.M., Awad, H.A., and Guilak, F. Influence of oxygen on the proliferation and metabolism of adipose derived adult stem cells. J Cell Physiol 204, 184, 2005.
44. Meyer, E.G., Buckley, C.T., Thorpe, S.D., and Kelly, D.J. Low oxygen tension is a more potent promoter of chondrogenic differentiation than dynamic compression. J Biomech 43, 2516, 2010.
45. Kanichai, M., Ferguson, D., Prendergast, P.J., and Campbell, V.A. Hypoxia promotes chondrogenesis in rat mesenchymal stem cells: A role for AKT and hypoxia-inducible factor (HIF)-1? J Cell Physiol 216, 708, 2008.
46. Robins, J.C., Akeno, N., Mukherjee, A., Dalal, R.R., Aronow, B.J., Koopman, P., et al. Hypoxia induces chondrocytespecific gene expression in mesenchymal cells in association with transcriptional activation of Sox9. Bone 37, 313, 2005.
47. Hirao, M., Tamai, N., Tsumaki, N., Yoshikawa, H., and Myoui, A. Oxygen tension regulates chondrocyte differentiation and function during endochondral ossification. J Biol Chem 281, 31079, 2006.
48. Pilgaard, L., Lund, P., Duroux, M., Fink, T., Ulrich-Vinther, M., Soballe, K., et al. Effect of oxygen concentration, culture format and donor variability on in vitro chondrogenesis of human adipose tissue-derived stem cells. Regen Med 4, 539, 2009.
49. Malladi, P., Xu, Y., Chiou, M., Giaccia, A.J., and Longaker, M.T. Effect of reduced oxygen tension on chondrogenesis and osteogenesis in adipose-derived mesenchymal cells. Am J Physiol Cell Physiol 290, C1139-46. 2006.
50. Khan, W.S., Adesida, A.B., and Hardingham, T.E. Hypoxic conditions increase hypoxia-inducible transcription factor 2alpha and enhance chondrogenesis in stem cells from the infrapatellar fat pad of osteoarthritis patients. Arthritis Res Ther 9, R55. 2007.
51. Buckley, C.T., Vinardell, T., and Kelly, D.J. Oxygen tension differentially regulates the functional properties of cartilaginous tissues engineered from infrapatellar fat pad derived MSCs and articular chondrocytes. Osteoarthritis Cartilage 18, 1345, 2010.
52. Zhou, S., Cui, Z., and Urban, J.P. Factors influencing the oxygen concentration gradient from the synovial surface of articular cartilage to the cartilage-bone interface: a modeling study. Arthritis Rheum 50, 3915, 2004.
53. Williams, 3rd, R.J., and Harnly, H.W. Microfracture: indications, technique, and results. Instr Course Lect 56, 419, 2007.
54. McIlwraith, C.W., and Frisbie, D.D. Microfracture: basic science studies in the horse. Cartilage 1, 87, 2010.
55. Digirolamo, C.M., Stokes, D., Colter, D., Phinney, D.G., Class, R., and Prockop, D.J. Propagation and senescence of human marrow stromal cells in culture: a simple colonyforming assay identifies samples with the greatest potential to propagate and differentiate. Br J Haematol 107, 275, 1999.
56. Bruder, S.P., Jaiswal, N., and Haynesworth, S.E. Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation. J Cell Biochem 64, 278, 1997.
57. Siddappa, R., Licht, R., van Blitterswijk, C., and de Boer, J. Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering. J Orthop Res 25, 1029, 2007.
58. Liu, Y., Buckley, C.T., Downey, R., Mulhall, K.J., and Kelly, D.J. The role of environmental factors in regulating the development of cartilaginous grafts engineered using osteoarthritic human infrapatellar fat pad derived stem cells. Tissue Eng Part A 2012 [Epub ahead of print]; DOI: 10.1089/ten. TEA.2011.0575.
59. Ahmed TAE, Dare, E.V., and Hincke, M. Fibrin: a versatile scaffold for tissue engineering applications. Tissue Eng Part B 14, 199, 2008.
60. Kelly, D.J., and Prendergast, P.J. Mechano-regulation of stem cell differentiation and tissue regeneration in osteochondral defects. J Biomech 38, 1413, 2005.
61. Kelly, D.J., and Prendergast, P.J. Prediction of the optimal mechanical properties for a scaffold used in osteochondral defect repair. Tissue Eng 12, 2509, 2006.
62. Steck, E., Fischer, J., Lorenz, H., Gotterbarm, T., Jung, M., and Richter, W. Mesenchymal stem cell differentiation in an experimental cartilage defect: restriction of hypertrophy to bone-close neocartilage. Stem Cells Dev 18, 969, 2009.
63. Kelly, D.J., and Jacobs, C.R. The role of mechanical signals in regulating chondrogenesis and osteogenesis of mesenchymal stem cells. Birth Defects Res Part C Embryo Today 90, 75, 2010.
64. Connelly, J.T., Vanderploeg, E.J., Mouw, J.K., Wilson, C.G., and Levenston, M.E. Tensile loading modulates bone marrow stromal cell differentiation and the development of engineered fibrocartilage constructs. Tissue Eng Part A 16, 1913, 2010.
65. Li, Z., Yao, S.J., Alini, M., and Stoddart, M.J. Chondrogenesis of human bone marrow mesenchymal stem cells in fibrinpolyurethane composites is modulated by frequency and amplitude of dynamic compression and shear stress. Tissue Eng Part A 16, 575, 2010.
66. Huang, C.Y., Hagar, K.L., Frost, L.E., Sun, Y., and Cheung, H.S. Effects of cyclic compressive loading on chondrogenesis of rabbit bone-marrow derived mesenchymal stem cells. Stem Cells 22, 313, 2004.
67. Thorpe, S.D., Buckley, C.T., Vinardell, T., O'Brien, F.J., Campbell, V.A., and Kelly, D.J. The response of bone marrow-derived mesenchymal stem cells to dynamic compression following TGF-b3 induced chondrogenic differentiation. Ann Biomed Eng 1, 2010.
68. Kisiday, J.D., Frisbie, D.D., McIlwraith, C.W., and Grodzinsky, A.J. Dynamic compression stimulates proteoglycan synthesis by mesenchymal stem cells in the absence of chondrogenic cytokines. Tissue Eng Part A 15, 2817, 2009.
69. Mouw, J.K., Connelly, J.T., Wilson, C.G., Michael, K.E., and Levenston, M.E. Dynamic compression regulates the expression and synthesis of chondrocyte-specific matrix molecules in bone marrow stromal cells. Stem Cells 25, 655, 2007.
70. Miyanishi, K., Trindade, M.C., Lindsey, D.P., Beaupre, G.S., Carter, D.R., Goodman, S.B., et al. Dose-and time-dependent effects of cyclic hydrostatic pressure on transforming growth factor-beta3-induced chondrogenesis by adult human mesenchymal stem cells in vitro. Tissue Eng 12, 2253, 2006.
71. Miyanishi, K., Trindade, M.C.D., Lindsey, D.P., Beaupre, G.S., Carter, D.R., Goodman, S.B., et al. Effects of hydrostatic pressure and transforming growth factor-beta 3 on adult human mesenchymal stem cell chondrogenesis in vitro. Tissue Eng 12, 1419, 2006.
72. Angele, P., Yoo, J.U., Smith, C., Mansour, J., Jepsen, K.J., Nerlich, M., et al. Cyclic hydrostatic pressure enhances the chondrogenic phenotype of human mesenchymal progenitor cells differentiated in vitro. J Orthop Res 21, 451, 2003.
73. Vinardell, T., Rolfe, R.A., Buckley, C.T.,Meyer, E.G., Ahearne, M., Murphy, P., et al. Hydrostatic pressure acts to stabilise a chondrogenic phenotype in porcine joint tissue derived stem cells. Eur Cell Mater 23, 121; discussion 33, 2012.
74. Steward, A.J., Thorpe, S.D., Vinardell, T., Buckley, C.T., Wagner, D.R., and Kelly, D.J. Cell-matrix interactions regulate mesenchymal stem cell response to hydrostatic pressure. Acta Biomaterialia 8, 2153, 2012.
75. Thorpe, S.D., Buckley, C.T., Vinardell, T., O'Brien, F.J., Campbell, V.A., and Kelly, D.J. Dynamic compression can inhibit chondrogenesis of mesenchymal stem cells. Biochem Biophys Res Commun 377, 458, 2008.
76. Huang, A.H., Farrell, M.J., Kim, M., and Mauck, R.L. Longterm dynamic loading improves the mechanical properties of chondrogenic mesenchymal stem cell-laden hydrogels. Eur Cells Mater 19, 72, 2010.
77. Huang, A.H., Farrell, M.J., and Mauck, R.L. Mechanics and mechanobiology of mesenchymal stem cell-based engineered cartilage. J Biomech 43, 128, 2010.
78. Mauck, R.L., Soltz, M.A., Wang, C.C.B., Wong, D.D., Chao, P.H.G., Valhmu, W.B., et al. Functional tissue engineering of articular cartilage through dynamic loading of chondrocyteseeded agarose gels. J Biomech Eng 122, 252, 2000.
79. Meyer, E.G., Buckley, C.T., Steward, A.J., and Kelly, D.J. The effect of cyclic hydrostatic pressure on the functional development of cartilaginous tissues engineered using bone marrow derived mesenchymal stem cells. J Mech Behav Biomed Mater 4, 1257, 2011.
80. Warnawin, E., Burakowski, T., Gajewski,M., Radzikowska, A., Kornatka, A., Michalak, C., et al. Preservation of chondrogenic potential of mesenchymal stem cells isolated from osteoarthritic patients during proliferation in response to plateletderived growth factor (PDGF). Cent-Eur J Immunol 30, 26, 2005.
81. Lacey, D.C., Simmons, P.J., Graves, S.E., and Hamilton, J.A. Proinflammatory cytokines inhibit osteogenic differentiation from stem cells: implications for bone repair during inflammation. Osteoarthritis Cartilage 17, 735, 2009.
82. Lei, J., Cheng, L.M., and Tan, M.Q. Effects of serum and cytokines on the proliferation of human marrow mesenchymal stem cells. Bull Hunan Med Univ 28, 469, 2003.
83. Heldens, G.T.H., Blaney Davidson, E.N., Vitters, E.L., Schreurs, B.W., Piek, E., Van Den Berg, W.B., et al. Catabolic factors and osteoarthritis- conditioned medium inhibit chondrogenesis of human mesenchymal stem cells. Tissue Eng Part A 18, 45, 2012.
84. Felka, T., Schäfer, R., Schewe, B., Benz, K., and Aicher, W.K. Hypoxia reduces the inhibitory effect of IL-1b on chondrogenic differentiation of FCS-free expanded MSC. Osteoarthritis Cartilage 17, 1368, 2009.
85. Majumdar, M.K., Wang, E., and Morris, E.A. BMP-2 and BMP-9 promote chondrogenic differentiation of human multipotential mesenchymal cells and overcome the inhibitory effect of IL-1. J Cell Physiol 189, 275, 2001.
86. Wehling,N., Palmer,G.D., Pilapil, C., Liu, F., Wells, J.W.,Müller, P.E., et al. Interleukin-1b and tumor necrosis factor a inhibit chondrogenesis by human mesenchymal stem cells through NF-kB-dependent pathways. Arthritis Rheum 60, 801, 2009.