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Asporin and transforming growth factor-β gene expression in osteoblasts from subchondral bone and osteophytes in osteoarthritis

  • Original Article
  • Published:
Journal of Orthopaedic Science

Abstract

Background

To clarify the significance of subchondral bone and osteophytes in the pathology of osteoarthritis (OA), we investigated the expression of asporin (ASPN), transforming growth factor-β1 (TGF-β1), TGF-β2, TGF-β3, and runt-related transcription factor-2 (Runx2) genes involved in bone metabolism.

Methods

Osteoblasts were isolated from 19 patients diagnosed with knee OA and from 4 patients diagnosed with femoral neck fracture. Osteoblast expression of mRNA encoding ASPN, TGF-β1, TGF-β2, TGF-β3, and Runx2 was analyzed using real-time RT-PCR.

Results

Expression of ASPN, TGF-β1, and TGF-β3 mRNA in the subchondral bone and osteophytes of OA patients increased compared with that of non-OA patients. The ratio of ASPN to TGF-β1 mRNA in patients with severe cartilage damage was higher than that in patients with mild cartilage damage.

Conclusions

The increased ratio of ASPN mRNA to TGF-β1 mRNA in patients with severe relative to mild cartilage damage indicates that increased ASPN mRNA expression was significantly associated with the severity of cartilage degeneration. This finding suggests that ASPN may regulate TGF-β1-mediated factors in the development of OA, which may provide clues as to the underlying pathology of OA.

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References

  1. Bailey AJ, Mansell JP. Do subchondral bone changes exacerbate or precede articular cartilage destruction in osteoarthritis of the elderly? Gerontology 1997;43:296–304.

    Article  CAS  PubMed  Google Scholar 

  2. Wei HW, Sun SS, Jao SH, Yeh CR, Cheng CK. The influence of mechanical properties of subchondral plate, femoral head and neck on dynamic stress distribution of the articular cartilage. Med Eng Phys 2005;27:295–304.

    Article  PubMed  Google Scholar 

  3. Diehl P, Schmitt M, Schauwecker J, Eichelberg K, Gollwitzer H, Gradinger R, et al. Effect of high hydrostatic pressure on biological properties of extracellular bone matrix proteins. Int J Mol Med 2005;16:285–289.

    CAS  PubMed  Google Scholar 

  4. Bosetti M, Boccafoschi F, Leigheb M, Cannas MF. Effect of different growth factors on human osteoblasts activities: a possible application in bone regeneration for tissue engineering. Biomol Eng 2007;24:613–618.

    Article  CAS  PubMed  Google Scholar 

  5. Marie PJ. Transcription factors controlling osteoblastogenesis. Arch Biochem Biophys 2008;473:98–105.

    Article  CAS  PubMed  Google Scholar 

  6. Lieb E, Vogel T, Milz S, Dauner M, Schulz MB. Effects of transforming growth factor beta-1 on bonelike tissue formation in three-dimensional cell culture. II: Osteoblastic differentiation. Tissue Eng 2004;10:1414–1425.

    CAS  PubMed  Google Scholar 

  7. Borton AJ, Frederick JP, Datto MB, Wang XF, Weinstein RS. The loss of Smad3 results in a lower rate of bone formation and osteopenia through dysregulation of osteoblast differentiation and apoptosis. J Bone Miner Res 2001;16:1754–1764.

    Article  CAS  PubMed  Google Scholar 

  8. Henry SP, Takanosu M, Boyd TC, Mayne PM, Eberspaecher H, Zhou W, et al. Expression pattern and gene characterization of asporin. a newly discovered member of the leucine-rich repeat protein family. J Biol Chem 2001;276:12212–12221.

    Article  CAS  PubMed  Google Scholar 

  9. Lorenzo P, Aspberg A, Onnerfjord P, Bayliss MT, Neame PJ, Heinegard D. Identification and characterization of asporin: a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan. J Biol Chem 2001;276:12201–12211.

    Article  CAS  PubMed  Google Scholar 

  10. Kizawa H, Kou I, Iida A, Sudo A, Miyamoto Y, Fukuda A, et al. An aspartic acid repeat polymorphism in asporin inhibits chondrogenesis and increases susceptibility to osteoarthritis. Nat Genet 2005;37:138–144.

    Article  CAS  PubMed  Google Scholar 

  11. Kamekura S, Kawasaki Y, Hoshi K, Shimoaka T, Chikuda H, Maruyama Z, et al. Contribution of runt-related transcription factor 2 to the pathogenesis of osteoarthritis in mice after induction of knee joint instability. Arthritis Rheum 2006;54:2462–2470.

    Article  CAS  PubMed  Google Scholar 

  12. Lee KS, Kim HJ, Li QL, Chi XZ, Ueta C, Komori T, et al. Runx2 is a common target of transforming growth factor beta 1 and bone morphogenetic protein 2, and cooperation between Runx2 and Smad5 induces osteoblast-specific gene expression in the pluripotent mesenchymal precursor cell line C2C12. Mol Cell Biol 2000;20:8783–8792.

    Article  CAS  PubMed  Google Scholar 

  13. Kellgren JH, Lawrence JS. Radiological assessment of osteoarthrosis. Ann Rheum Dis 1957;16:494–502.

    Article  CAS  PubMed  Google Scholar 

  14. Kleemann RU, Krocker D, Cedraro A, Tuischer J, Duda GN. Altered cartilage mechanics and histology in knee osteoarthritis: relation to clinical assessment (ICRS grade). Osteoarthritis Cartilage 2005;13:958–963.

    Article  CAS  PubMed  Google Scholar 

  15. Beresford JN, Gallagher JA, Gowen M, Couch M, Poser J, Wood DD, Russell RG. The effects of monocyte-conditioned medium and interleukin 1 on the synthesis of collagenous and noncollagenous proteins by mouse bone and human bone cells in vitro. Biochim Biophys Acta 1984;801:58–65.

    CAS  PubMed  Google Scholar 

  16. Sakao K, Takahashi KA, Mazda O, Arai Y, Tonomura H, Inoue A, et al. Enhanced expression of interleukin-6, matrix metalloproteinase-13, and receptor activator of NF-κB ligand in cells derived from osteoarthritic subchondral bone. J Orthop Sci 2 2008;13:202–210.

    Article  CAS  Google Scholar 

  17. Sakao K, Takahashi KA, Arai Y, Saito M, Honjo K, Hiraoka N, et al. Osteoblasts derived from osteophytes produce interleukin-6, interleukin-8, and matrix metalloproteinase-13 in osteoarthritis. J Bone Miner Metab 2009;27:412–423.

    Article  CAS  PubMed  Google Scholar 

  18. Inoue A, Takahashi KA, Arai Y, Tonomura H, Sakao K, Saito M, et al. The therapeutic effects of basic fibroblast growth factor contained in gelatin hydrogel microspheres on experimental osteoarthritis in the rabbit knee. Arthritis Rheum 2006;54:264–270.

    Article  CAS  PubMed  Google Scholar 

  19. Bustin SA. Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol 2002;29:23–39.

    Article  CAS  PubMed  Google Scholar 

  20. Ameye L, Aria D, Jepsen K, Oldberg A, Xu T, Young MF. Abnormal collagen fibrils in tendons of biglycan/fibromodulin-deficient mice lead to gait impairment, ectopic ossification, and osteoarthritis. FASEB J 2002;16:673–680.

    Article  CAS  PubMed  Google Scholar 

  21. Gill MR, Oldberg A, Reinholt FP. Fibromodulin-null murine knee joints display increased incidences of osteoarthritis and alterations in tissue biochemistry. Osteoarthritis Cartilage 2002;10:751–757.

    Article  CAS  PubMed  Google Scholar 

  22. Nakajima M, Kizawa H, Saitoh M, Kou I, Miyazono K, Ikegawa S. Mechanisms for asporin function and regulation in articular cartilage. J Biol Chem 2007;282:32185–32192.

    Article  CAS  PubMed  Google Scholar 

  23. Ikegawa S, Kawamura S, Takahashi A, Nakamura T, Kamatani N. Replication of association of the D-repeat polymorphism in asporin with osteoarthritis. Arthritis Res Ther 2006;8:403; author reply 403.

    Article  PubMed  Google Scholar 

  24. Jiang Q, Shi D, Yi L, Ikegawa S, Wang Y, Nakamura T, et al. Replication of the association of the aspartic acid repeat polymorphism in the asporin gene with knee-osteoarthritis susceptibility in Han Chinese. J Hum Genet 2006;51:1068–1072.

    Article  CAS  PubMed  Google Scholar 

  25. Yang X, Chen L, Xu X, Li C, Huang C, Deng CX. TGF-beta/Smad3 signals repress chondrocyte hypertrophic differentiation and are required for maintaining articular cartilage. J Cell Biol 2001;153:35–46.

    Article  CAS  PubMed  Google Scholar 

  26. Blaney Davidson EN, Vitters EL, van der Kraan PM, van den Berg WB. Expression of transforming growth factor-beta (TGFbeta) and the TGFbeta signalling molecule SMAD-2P in spontaneous and instability-induced osteoarthritis: role in cartilage degradation, chondrogenesis and osteophyte formation. Ann Rheum Dis 2006;65:1414–1421.

    Article  CAS  PubMed  Google Scholar 

  27. Scharstuhl A, Glansbeek HL, van Beuningen HM, Vitters EL, van der Kraan PM, van den Berg WB. Inhibition of endogenous TGF-beta during experimental osteoarthritis prevents osteophyte formation and impairs cartilage repair. J Immunol 2002;169:507–514.

    CAS  PubMed  Google Scholar 

  28. Scharstuhl A, Vitters EL, van der Kraan PM, van den Berg WB. Reduction of osteophyte formation and synovial thickening by adenoviral overexpression of transforming growth factor beta/bone morphogenetic protein inhibitors during experimental osteoarthritis. Arthritis Rheum 2003;48:3442–3451.

    Article  CAS  PubMed  Google Scholar 

  29. Okazaki K, Jingushi S, Ikenoue T, Urabe K, Sakai H, Ohtsuru A, et al. Expression of insulin-like growth factor I messenger ribonucleic acid in developing osteophytes in murine experimental osteoarthritis and in rats inoculated with growth hormone-secreting tumor. Endocrinology 1999;140:4821–4830.

    Article  CAS  PubMed  Google Scholar 

  30. van Beuningen HM, Glansbeek HL, van der Kraan PM, van den Berg WB. Osteoarthritis-like changes in the murine knee joint resulting from intra-articular transforming growth factor-beta injections. Osteoarthritis Cartilage 2000;8:25–33.

    Article  PubMed  Google Scholar 

  31. Arai Y, Kubo T, Kobayashi K, Takeshita K, Takahashi K, Ikeda T, et al. Adenovirus vector-mediated gene transduction to chondrocytes: in vitro evaluation of therapeutic efficacy of transforming growth factor-beta 1 and heat shock protein 70 gene transduction. J Rheumatol 1997;24:1787–1795.

    CAS  PubMed  Google Scholar 

  32. Grimaud E, Heymann D, Redini F. Recent advances in TGF-beta effects on chondrocyte metabolism. Potential therapeutic roles of TGF-beta in cartilage disorders. Cytokine Growth Factor Rev 2002;13:241–257.

    Article  CAS  PubMed  Google Scholar 

  33. Shuler FD, Georgescu HI, Niyibizi C, Studer RK, Mi Z, Johnstone B, et al. Increased matrix synthesis following adenoviral transfer of a transforming growth factor beta-1 gene into articular chondrocytes. J Orthop Res 2000;18:585–592.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Orthopaedics, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto, 602-8566, Japan

    Kei Sakao, Kenji A. Takahashi, Yuji Arai, Masazumi Saito, Kuniaki Honjyo, Nobuyuki Hiraoka & Toshikazu Kubo

  2. Department of Microbiology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan

    Tsunao Kishida, Osam Mazda & Jiro Imanishi

Authors
  1. Kei Sakao
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  2. Kenji A. Takahashi
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  3. Yuji Arai
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  4. Masazumi Saito
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  5. Kuniaki Honjyo
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  6. Nobuyuki Hiraoka
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  7. Tsunao Kishida
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  8. Osam Mazda
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  9. Jiro Imanishi
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  10. Toshikazu Kubo
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Sakao, K., Takahashi, K.A., Arai, Y. et al. Asporin and transforming growth factor-β gene expression in osteoblasts from subchondral bone and osteophytes in osteoarthritis. J Orthop Sci 14, 738–747 (2009). https://doi.org/10.1007/s00776-009-1401-4

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  • DOI: https://doi.org/10.1007/s00776-009-1401-4

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