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Cartilage Matrix Formation by Bovine Mesenchymal Stem Cells in Three-dimensional Culture Is Age-dependent

  • Symposium: Clinically Relevant Strategies for Treating Cartilage and Meniscal Pathology
  • Published:
Clinical Orthopaedics and Related Research®

Abstract

Background

Cartilage degeneration is common in the aged, and aged chondrocytes are inferior to juvenile chondrocytes in producing cartilage-specific extracellular matrix. Mesenchymal stem cells (MSCs) are an alternative cell type that can differentiate toward the chondrocyte phenotype. Aging may influence MSC chondrogenesis but remains less well studied, particularly in the bovine system.

Questions/purposes

The objectives of this study were (1) to confirm age-related changes in bovine articular cartilage, establish how age affects chondrogenesis in cultured pellets for (2) chondrocytes and (3) MSCs, and (4) determine age-related changes in the biochemical and biomechanical development of clinically relevant MSC-seeded hydrogels.

Methods

Native bovine articular cartilage from fetal (n = 3 donors), juvenile (n = 3 donors), and adult (n = 3 donors) animals was analyzed for mechanical and biochemical properties (n = 3–5 per donor). Chondrocyte and MSC pellets (n = 3 donors per age) were cultured for 6 weeks before analysis of biochemical content (n = 3 per donor). Bone marrow-derived MSCs of each age were also cultured within hyaluronic acid hydrogels for 3 weeks and analyzed for matrix deposition and mechanical properties (n = 4 per age).

Results

Articular cartilage mechanical properties and collagen content increased with age. We observed robust matrix accumulation in three-dimensional pellet culture by fetal chondrocytes with diminished collagen-forming capacity in adult chondrocytes. Chondrogenic induction of MSCs was greater in fetal and juvenile cell pellets. Likewise, fetal and juvenile MSCs in hydrogels imparted greater matrix and mechanical properties.

Conclusions

Donor age and biochemical microenvironment were major determinants of both bovine chondrocyte and MSC functional capacity.

Clinical Relevance

In vitro model systems should be evaluated in the context of age-related changes and should be benchmarked against human MSC data.

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References

  1. Adkisson HD, Gillis MP, Davis EC, Maloney W, Hruska KA. In vitro generation of scaffold independent neocartilage. Clin Orthop Relat Res. 2001;391(Suppl):S280–294.

    Article  PubMed  Google Scholar 

  2. Archer CW, Dowthwaite GP, Francis-West P. Development of synovial joints. Birth Defects Res C Embryo Today. 2003;69:144–155.

    Article  PubMed  CAS  Google Scholar 

  3. Ateshian GA, Hung CT. Patellofemoral joint biomechanics and tissue engineering. Clin Orthop Relat Res. 2005;436:81–90.

    Article  PubMed  Google Scholar 

  4. Baksh D, Song L, Tuan RS. Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med. 2004;8:301–316.

    Article  PubMed  CAS  Google Scholar 

  5. Barbero A, Grogan S, Schafer D, Heberer M, Mainil-Varlet P, Martin I. Age related changes in human articular chondrocyte yield, proliferation and post-expansion chondrogenic capacity. Osteoarthritis Cartilage. 2004;12:476–484.

    Article  PubMed  Google Scholar 

  6. Barry F, Boynton RE, Liu B, Murphy JM. Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. Exp Cell Res. 2001;268:189–200.

    Article  PubMed  CAS  Google Scholar 

  7. Below S, Arnoczky SP, Dodds J, Kooima C, Walter N. The split-line pattern of the distal femur: a consideration in the orientation of autologous cartilage grafts. Arthroscopy. 2002;18:613–617.

    Article  PubMed  Google Scholar 

  8. Bernardo ME, Emons JAM, Karperien M, Nauta AJ, Willemze R, Roelofs H, Romeo S, Marchini A, Rappold GA, Vukicevic S, Locatelli F, Fibbe WE. Human mesenchymal stem cells derived from bone marrow display a better chondrogenic differentiation compared with other sources. Connect Tissue Res. 2007;48:132–140.

    Article  PubMed  CAS  Google Scholar 

  9. Bullough P, Goodfellow J. The significance of the fine structure of articular cartilage. J Bone Joint Surg Br. 1968;50:852–857.

    PubMed  CAS  Google Scholar 

  10. Burdick JA, Chung C, Jia X, Randolph MA, Langer R. Controlled degradation and mechanical behavior of photopolymerized hyaluronic acid networks. Biomacromolecules. 2005;6:386–391.

    Article  PubMed  CAS  Google Scholar 

  11. Charlebois M, McKee MD, Buschmann MD. Nonlinear tensile properties of bovine articular cartilage and their variation with age and depth. J Biomech Eng. 2004;126:129–137.

    Article  PubMed  Google Scholar 

  12. Chung C, Burdick JA. Influence of three-dimensional hyaluronic acid microenvironments on mesenchymal stem cell chondrogenesis. Tissue Eng Part A. 2009;15:243–254.

    Article  PubMed  CAS  Google Scholar 

  13. Clarke IC. Articular cartilage: a review and scanning electron microscope study. 1. The interterritorial fibrillar architecture. J Bone Joint Surg Br. 1971;53:732–750.

    PubMed  CAS  Google Scholar 

  14. Coipeau P, Rosset P, Langonne A, Gaillard J, Delorme B, Rico A, Domenech J, Charbord P, Sensebe L. Impaired differentiation potential of human trabecular bone mesenchymal stromal cells from elderly patients. Cytotherapy. 2009;11:584–594.

    Article  PubMed  CAS  Google Scholar 

  15. Detterline AJ, Goldberg S, Bach BR Jr, Cole BJ. Treatment options for articular cartilage defects of the knee. Orthop Nurs. 2005;24:361–366; quiz 367–368.

    Google Scholar 

  16. Dressler MR, Butler DL, Boivin GP. Effects of age on the repair ability of mesenchymal stem cells in rabbit tendon. J Orthop Res. 2005;23:287–293.

    Article  PubMed  CAS  Google Scholar 

  17. Erickson IE, Huang AH, Chung C, Li RT, Burdick JA, Mauck RL. Differential maturation and structure-function relationships in mesenchymal stem cell- and chondrocyte-seeded hydrogels. Tissue Eng Part A. 2009;15:1041–1052.

    Article  PubMed  CAS  Google Scholar 

  18. Erickson IE, Huang AH, Sengupta S, Kestle S, Burdick JA, Mauck RL. Macromer density influences mesenchymal stem cell chondrogenesis and maturation in photocrosslinked hyaluronic acid hydrogels. Osteoarthritis Cartilage. 2009;17:1639–1648.

    Article  PubMed  CAS  Google Scholar 

  19. Farndale RW, Buttle DJ, Barrett AJ. Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta. 1986;883:173–177.

    PubMed  CAS  Google Scholar 

  20. Frankowski JJ, Watkins-Castillo S. Primary Total Knee and Hip Arthroplasty Projections for the US Population to the Year 2030. Rosemont, IL: American Academy of Orthopaedic Surgeons, Department of Research and Scientific Affairs; 2002:1–8.

    Google Scholar 

  21. Giannoni P, Pagano A, Maggi E, Arbico R, Randazzo N, Grandizio M, Cancedda R, Dozin B. Autologous chondrocyte implantation (ACI) for aged patients: development of the proper cell expansion conditions for possible therapeutic applications. Osteoarthritis Cartilage. 2005;13:589–600.

    Article  PubMed  CAS  Google Scholar 

  22. Huang AH, Farrell MJ, Kim M, Mauck RL. Long-term dynamic loading improves the mechanical properties of chondrogenic mesenchymal stem cell-laden hydrogel. Eur Cell Mater. 2010;19:72–85.

    PubMed  CAS  Google Scholar 

  23. Huang AH, Stein A, Mauck RL. Evaluation of the complex transcriptional topography of mesenchymal stem cell chondrogenesis for cartilage tissue engineering. Tissue Eng Part A. 2010;16:2699–2708.

    Article  PubMed  CAS  Google Scholar 

  24. Huang AH, Stein A, Tuan RS, Mauck RL. Transient exposure to transforming growth factor beta 3 improves the mechanical properties of mesenchymal stem cell-laden cartilage constructs in a density-dependent manner. Tissue Eng Part A. 2009;15:3461–3472.

    Article  PubMed  CAS  Google Scholar 

  25. Huang AH, Yeger-McKeever M, Stein A, Mauck RL. Tensile properties of engineered cartilage formed from chondrocyte- and MSC-laden hydrogels. Osteoarthritis Cartilage. 2008;16:1074–1082.

    Article  PubMed  CAS  Google Scholar 

  26. Kempson GE. Age-related changes in the tensile properties of human articular cartilage: a comparative study between the femoral head of the hip joint and the talus of the ankle joint. Biochim Biophys Acta. 1991;1075:223–230.

    PubMed  CAS  Google Scholar 

  27. Kleemann RU, Schell H, Thompson M, Epari DR, Duda GN, Weiler A. Mechanical behavior of articular cartilage after osteochondral autograft transfer in an ovine model. Am J Sports Med. 2007;35:555–563.

    Article  PubMed  Google Scholar 

  28. Knutsen G, Engebretsen L, Ludvigsen TC, Drogset JO, Grontvedt T, Solheim E, Strand T, Roberts S, Isaksen V, Johansen O. Autologous chondrocyte implantation compared with microfracture in the knee. A randomized trial. J Bone Joint Surg Am. 2004;86:455–464.

    PubMed  Google Scholar 

  29. Kopesky PW, Lee HY, Vanderploeg EJ, Kisiday JD, Frisbie DD, Plaas AH, Ortiz C, Grodzinsky AJ. Adult equine bone marrow stromal cells produce a cartilage-like ECM mechanically superior to animal-matched adult chondrocytes. Matrix Biol. 2010;29:427–438.

    Article  PubMed  CAS  Google Scholar 

  30. Kretlow J, Jin Y-Q, Liu W, Zhang W, Hong T-H, Zhou G, Baggett LS, Mikos A, Cao Y. Donor age and cell passage affects differentiation potential of murine bone marrow-derived stem cells. BMC Cell Biology. 2008;9:60.

    Article  PubMed  Google Scholar 

  31. Martin JA, Buckwalter JA. Telomere erosion and senescence in human articular cartilage chondrocytes. J Gerontol A Biol Sci Med Sci. 2001;56:B172–179.

    Article  PubMed  CAS  Google Scholar 

  32. Mauck RL, Soltz MA, Wang CC, Wong DD, Chao PH, Valhmu WB, Hung CT, Ateshian GA. Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels. J Biomech Eng. 2000;122:252–260.

    Article  PubMed  CAS  Google Scholar 

  33. Mauck RL, Yuan X, Tuan RS. Chondrogenic differentiation and functional maturation of bovine mesenchymal stem cells in long-term agarose culture. Osteoarthritis Cartilage. 2006;14:179–189.

    Article  PubMed  CAS  Google Scholar 

  34. Micheli L, Curtis C, Shervin N. Articular cartilage repair in the adolescent athlete: is autologous chondrocyte implantation the answer? Clin J Sport Med. 2006;16:465–470.

    Article  PubMed  Google Scholar 

  35. Micheli LJ, Browne JE, Erggelet C, Fu F, Mandelbaum B, Moseley JB, Zurakowski D. Autologous chondrocyte implantation of the knee: multicenter experience and minimum 3-year follow-up. Clin J Sport Med. 2001;11:223–228.

    Article  PubMed  CAS  Google Scholar 

  36. Morrison EH, Ferguson MW, Bayliss MT, Archer CW. The development of articular cartilage: I. The spatial and temporal patterns of collagen types. J Anat. 1996;189:9–22.

    PubMed  CAS  Google Scholar 

  37. Murphy JM, Dixon K, Beck S, Fabian D, Feldman A, Barry F. Reduced chondrogenic and adipogenic activity of mesenchymal stem cells from patients with advanced osteoarthritis. Arthritis Rheum. 2002;46:704–713.

    Article  PubMed  Google Scholar 

  38. Neuman RE, Logan MA. The determination of hydroxypoline. J Biol Chem. 1950;184:299–306.

    PubMed  CAS  Google Scholar 

  39. Ng KW, Lima EG, Bian L, O’Conor CJ, Jayabalan PS, Stoker AM, Kuroki K, Cook CR, Ateshian GA, Cook JL, Hung CT. Passaged adult chondrocytes can form engineered cartilage with functional mechanical properties: a canine model. Tissue Eng Part A. 2010;16:1041–1051.

    Article  PubMed  CAS  Google Scholar 

  40. Park S, Nicoll S, Mauck R, Ateshian G. Cartilage mechanical response under dynamic compression at physiological stress levels following collagenase digestion. Ann Biomed Eng. 2008;36:425–434.

    Article  PubMed  Google Scholar 

  41. Payne KA, Didiano DM, Chu CR. Donor sex and age influence the chondrogenic potential of human femoral bone marrow stem cells. Osteoarthritis Cartilage. 2010;18:705–713.

    Article  PubMed  CAS  Google Scholar 

  42. Roura S, Farre J, Soler-Botija C, Llach A, Hove-Madsen L, Cairo JJ, Godia F, Cinca J, Bayes-Genis A. Effect of aging on the pluripotential capacity of human CD105(+) mesenchymal stem cells. Eur J Heart Fail. 2006:555–563.

  43. Scharstuhl A, Schewe B, Benz K, Gaissmaier C, Bühring H-J, Stoop R. Chondrogenic potential of human adult mesenchymal stem cells is independent of age or osteoarthritis etiology. Stem Cells. 2007;25:3244–3251.

    Article  PubMed  CAS  Google Scholar 

  44. Steadman JR, Rodkey WG, Rodrigo JJ. Microfracture: surgical technique and rehabilitation to treat chondral defects. Clin Orthop Relat Res. 2001;391(Suppl):S362–369.

    Article  PubMed  Google Scholar 

  45. Stegemann H, Stalder K. Determination of hydroxyproline. Clin Chim Acta. 1967;18:267–273.

    Article  PubMed  CAS  Google Scholar 

  46. Stenderup K, Justesen J, Clausen C, Kassem M. Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. Bone. 2003;33:919–926.

    Article  PubMed  Google Scholar 

  47. Stolzing A, Jones E, McGonagle D, Scutt A. Age-related changes in human bone marrow-derived mesenchymal stem cells: consequences for cell therapies. Mechanisms of Ageing and Development. 2008;129:163–173.

    Article  PubMed  CAS  Google Scholar 

  48. Temple MM, Bae WC, Chen MQ, Lotz M, Amiel D, Coutts RD, Sah RL. Age- and site-associated biomechanical weakening of human articular cartilage of the femoral condyle. Osteoarthritis Cartilage. 2007;15:1042–1052.

    Article  PubMed  CAS  Google Scholar 

  49. Tokalov SV, Gruner S, Schindler S, Wolf G, Baumann M, Abolmaali N. Age-related changes in the frequency of mesenchymal stem cells in the bone marrow of rats. Stem Cells and Development. 2007;16:439–446.

    Article  PubMed  CAS  Google Scholar 

  50. Tran-Khanh N, Hoemann CD, McKee MD, Henderson JE, Buschmann MD. Aged bovine chondrocytes display a diminished capacity to produce a collagen-rich, mechanically functional cartilage extracellular matrix. J Orthop Res. 2005;23:1354–1362.

    Article  PubMed  CAS  Google Scholar 

  51. Williamson AK, Chen AC, Masuda K, Thonar EJ, Sah RL. Tensile mechanical properties of bovine articular cartilage: variations with growth and relationships to collagen network components. J Orthop Res. 2003;21:872–880.

    Article  PubMed  CAS  Google Scholar 

  52. Williamson AK, Chen AC, Sah RL. Compressive properties and function-composition relationships of developing bovine articular cartilage. J Orthop Res. 2001;19:1113–1121.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgment

We thank Dr Jason A. Burdick for helpful discussions regarding this work.

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Authors and Affiliations

  1. McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, University of Pennsylvania, 424 Stemmler Hall, 36th Street and Hamilton Walk, Philadelphia, PA, 19104, USA

    Isaac E. Erickson BS, Steven C. van Veen, Swarnali Sengupta, Sydney R. Kestle & Robert L. Mauck PhD

  2. Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA

    Isaac E. Erickson BS, Sydney R. Kestle & Robert L. Mauck PhD

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  1. Isaac E. Erickson BS
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  2. Steven C. van Veen
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  3. Swarnali Sengupta
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  4. Sydney R. Kestle
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  5. Robert L. Mauck PhD
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Corresponding author

Correspondence to Robert L. Mauck PhD.

Additional information

One or more of the authors received funding from the National Institutes of Health (RO3 AR053668 and RO1 EB008722 [RLM]), the Penn Center for Musculoskeletal Disorders (AR050950 [RLM]), the Penn Institute on Aging (RLM), and the National Science Foundation (IEE). Additional support was from an NSF-sponsored REU program through the Nano-Bio Interface Center (NBIC) at the University of Pennsylvania (SS).

This work was performed at the University of Pennsylvania, Philadelphia, PA, USA.

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Erickson, I.E., van Veen, S.C., Sengupta, S. et al. Cartilage Matrix Formation by Bovine Mesenchymal Stem Cells in Three-dimensional Culture Is Age-dependent. Clin Orthop Relat Res 469, 2744–2753 (2011). https://doi.org/10.1007/s11999-011-1869-z

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  • DOI: https://doi.org/10.1007/s11999-011-1869-z

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