Annals of Biomedical Engineering

, Volume 46, Issue 6, pp 810–818 | Cite as

Cartilage Metabolism is Modulated by Synovial Fluid Through Metalloproteinase Activity

  • Eric Y. Sun
  • Allison K. M. Fleck
  • Ahmad E. Abu-Hakmeh
  • Alexandra Kotsakis
  • Garrett R. Leonard
  • Leo Q. Wan


Synovial fluid (SF) contains various cytokines that regulate chondrocyte metabolism and is dynamically associated with joint disease. The objective of this study was to investigate the effects of diluted normal SF on catabolic metabolism of articular cartilage under inflammatory conditions. For this purpose, SF was isolated from healthy bovine joints, diluted, and added to cartilage explant cultures stimulated with interleukin-1 (IL-1) for 12 days. The kinetic release of sulfated glycosaminoglycan (sGAG) and collagen, as well as nitric oxide and gelatinase matrix metalloproteinases were analyzed in the supernatant. Chondrocyte survival and matrix integrity in the explants were evaluated with Live/Dead and histological staining. Diluted synovial fluid treatment suppressed sGAG and collagen release, downregulated the production of nitric oxide and matrix metalloproteinases, reduced IL-1-induced chondrocyte death, and rescued matrix depletion. Our results demonstrate that normal SF can counteract inflammation-driven cartilage catabolism. This study reports on the protective function of healthy SF and the therapeutic potential of recapitulation of SF for cartilage repair.


Articular cartilage Interleukin-1 (IL-1) Inflammation Matrix degradation Osteoarthritis 





Rheumatoid arthritis


Extracellular matrix


Nitric oxide


Matrix metalloproteinase


Synovial fluid


Hyaluronic acid


Sulfated glycosaminoglycan



Leo Q. Wan is a Pew Scholar in Biomedical Science, supported by Pew Charitable Trusts.

Conflict of interest

The authors declare no conflicts of interest.

Supplementary material

10439_2018_2010_MOESM1_ESM.pdf (2.3 mb)
Supplementary material 1 (PDF 2360 kb)


  1. 1.
    Aigner, T., S. Soeder, and J. Haag. IL-1beta and BMPs–interactive players of cartilage matrix degradation and regeneration. Eur. Cell Mater. 12:49–56, 2006.CrossRefPubMedGoogle Scholar
  2. 2.
    Aizawa, T., T. Kon, T. A. Einhorn, and L. C. Gerstenfeld. Induction of apoptosis in chondrocytes by tumor necrosis factor-alpha. J. Orthop. Res. 19:785–796, 2001.CrossRefPubMedGoogle Scholar
  3. 3.
    Alquraini, A., S. Garguilo, G. D’Souza, L. X. Zhang, T. A. Schmidt, G. D. Jay, and K. A. Elsaid. The interaction of lubricin/proteoglycan 4 (PRG4) with toll-like receptors 2 and 4: an anti-inflammatory role of PRG4 in synovial fluid. Arthritis Res. Ther. 17:353, 2015.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Arechavaleta-Velasco, F., D. Ogando, S. Parry, and F. Vadillo-Ortega. Production of matrix metalloproteinase-9 in lipopolysaccharide-stimulated human amnion occurs through an autocrine and paracrine proinflammatory cytokine-dependent system. Biol. Reprod. 67:1952–1958, 2002.CrossRefPubMedGoogle Scholar
  5. 5.
    Babykutty, S., P. Suboj, P. Srinivas, A. S. Nair, K. Chandramohan, and S. Gopala. Insidious role of nitric oxide in migration/invasion of colon cancer cells by upregulating MMP-2/9 via activation of cGMP-PKG-ERK signaling pathways. Clin. Exp. Metastasis 29:471–492, 2012.CrossRefPubMedGoogle Scholar
  6. 6.
    Bauvois, B. New facets of matrix metalloproteinases MMP-2 and MMP-9 as cell surface transducers: outside-in signaling and relationship to tumor progression. Biochim. Biophys. Acta 29–36:2012, 1825.Google Scholar
  7. 7.
    Beekhuizen, M., Y. M. Bastiaansen-Jenniskens, W. Koevoet, D. B. Saris, W. J. Dhert, L. B. Creemers, and G. J. van Osch. Osteoarthritic synovial tissue inhibition of proteoglycan production in human osteoarthritic knee cartilage: establishment and characterization of a long-term cartilage-synovium coculture. Arthritis Rheum. 63:1918–1927, 2011.CrossRefPubMedGoogle Scholar
  8. 8.
    Bian, L., E. G. Lima, S. L. Angione, K. W. Ng, D. Y. Williams, D. Xu, A. M. Stoker, J. L. Cook, G. A. Ateshian, and C. T. Hung. Mechanical and biochemical characterization of cartilage explants in serum-free culture. J. Biomech. 41:1153–1159, 2008.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Bird, T. A., and J. Saklatvala. Identification of a common class of high affinity receptors for both types of porcine interleukin-1 on connective tissue cells. Nature 324:263, 1986.CrossRefPubMedGoogle Scholar
  10. 10.
    Brand, J. A., T. E. McAlindon, and L. Zeng. A 3D system for culturing human articular chondrocytes in synovial fluid. J. Vis. Exp. 59:e3587, 2012.Google Scholar
  11. 11.
    Conde, J., M. Scotece, V. Abella, A. Lois, V. Lopez, T. Garcia-Caballero, J. Pino, J. J. Gomez-Reino, R. Gomez, F. Lago, and O. Gualillo. IL-36alpha: a novel cytokine involved in the catabolic and inflammatory response in chondrocytes. Sci. Rep. 5:16674, 2015.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Fietz, S., R. Einspanier, S. Hoppner, B. Hertsch, and A. Bondzio. Determination of MMP-2 and -9 activities in synovial fluid of horses with osteoarthritic and arthritic joint diseases using gelatin zymography and immunocapture activity assays. Equine Vet. J. 40:266–271, 2008.CrossRefPubMedGoogle Scholar
  13. 13.
    Fosang, A. J., K. Last, H. Stanton, S. B. Golub, C. B. Little, L. Brown, and D. C. Jackson. Neoepitope antibodies against MMP-cleaved and aggrecanase-cleaved aggrecan. In: Matrix Metalloproteinase Protocols, edited by I. M. Clark. Totowa, NJ: Humana Press, 2010, pp. 305–340.CrossRefGoogle Scholar
  14. 14.
    Hashizume, M., and M. Mihara. High molecular weight hyaluronic acid inhibits IL-6-induced MMP production from human chondrocytes by up-regulating the ERK inhibitor, MKP-1. Biochem. Biophys. Res. Commun. 403:184–189, 2010.CrossRefPubMedGoogle Scholar
  15. 15.
    Hegewald, A. A., J. Ringe, J. Bartel, I. Kruger, M. Notter, D. Barnewitz, C. Kaps, and M. Sittinger. Hyaluronic acid and autologous synovial fluid induce chondrogenic differentiation of equine mesenchymal stem cells: a preliminary study. Tissue Cell 36:431–438, 2004.CrossRefPubMedGoogle Scholar
  16. 16.
    Hewitt, R. E., M. L. Corcoran, and W. G. Stetler-Stevenson. The activation, expression and function of gelatinase A (MMP-2). Trends Glycosci. Glycotechnol. 8:23–36, 1996.CrossRefGoogle Scholar
  17. 17.
    Hidaka, C., M. Quitoriano, R. F. Warren, and R. G. Crystal. Enhanced matrix synthesis and in vitro formation of cartilage-like tissue by genetically modified chondrocytes expressing BMP-7. J. Orthop. Res. 19:751–758, 2001.CrossRefPubMedGoogle Scholar
  18. 18.
    Hoff, P., F. Buttgereit, G. R. Burmester, M. Jakstadt, T. Gaber, K. Andreas, G. Matziolis, C. Perka, and E. Rohner. Osteoarthritis synovial fluid activates pro-inflammatory cytokines in primary human chondrocytes. Int. Orthop. 37:145–151, 2013.CrossRefPubMedGoogle Scholar
  19. 19.
    Lakey, R. L., and T. E. Cawston. Sulfasalazine blocks the release of proteoglycan and collagen from cytokine stimulated cartilage and down-regulates metalloproteinases. Rheumatology (Oxford) 48:1208–1212, 2009.CrossRefGoogle Scholar
  20. 20.
    Larbre, J. P., A. R. Moore, J. A. Da Silva, H. Iwamura, Y. Ioannou, and D. A. Willoughby. Direct degradation of articular cartilage by rheumatoid synovial fluid: contribution of proteolytic enzymes. J. Rheumatol. 21:1796–1801, 1994.PubMedGoogle Scholar
  21. 21.
    Lee, D. A., V. Salih, E. F. Stockton, J. S. Stanton, and G. Bentley. Effect of normal synovial fluid on the metabolism of articular chondrocytes in vitro. Clin. Orthop. Relat. Res. 342:228–238, 1997.CrossRefGoogle Scholar
  22. 22.
    Liao, W., Z. Li, H. Zhang, J. Li, K. Wang, and Y. Yang. Proteomic analysis of synovial fluid as an analytical tool to detect candidate biomarkers for knee osteoarthritis. Int. J. Clin. Exp. Pathol. 8:9975–9989, 2015.PubMedPubMedCentralGoogle Scholar
  23. 23.
    Mahmoodi, M., S. Sahebjam, D. Smookler, R. Khokha, and J. S. Mort. Lack of tissue inhibitor of metalloproteinases-3 results in an enhanced inflammatory response in antigen-induced arthritis. Am. J. Pathol. 166:1733–1740, 2005.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Malyak, M., R. E. Swaney, and W. P. Arend. Levels of synovial fluid interleukin-1 receptor antagonist in rheumatoid arthritis and other arthropathies. potential contribution from synovial fluid neutrophils. Arthritis Rheum. 36:781–789, 1993.CrossRefPubMedGoogle Scholar
  25. 25.
    McNulty, A. L., N. E. Rothfusz, H. A. Leddy, and F. Guilak. Synovial fluid concentrations and relative potency of interleukin-1 alpha and beta in cartilage and meniscus degradation. J. Orthop. Res. 31:1039–1045, 2013.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Mueller, M. B., and R. S. Tuan. Anabolic/Catabolic balance in pathogenesis of osteoarthritis: identifying molecular targets. PM R 3:S3–11, 2011.CrossRefPubMedGoogle Scholar
  27. 27.
    Nuver-Zwart, I., J. Schalkwijk, L. A. Joosten, W. B. van den Berg, and L. B. van de Putte. Effects of synovial fluid and synovial fluid cells on chondrocyte metabolism in short term tissue culture. J. Rheumatol. 15:210–216, 1988.PubMedGoogle Scholar
  28. 28.
    Olszewska-Slonina, D., S. Jung, D. Matewski, K. J. Olszewski, E. Krzyzynska-Malinowska, A. Braszkiewicz, and B. Kowaliszyn. Lysosomal enzymes in serum and synovial fluid in patients with osteoarthritis. Scand. J. Clin. Lab. Invest. 75:145–151, 2015.CrossRefPubMedGoogle Scholar
  29. 29.
    Punzi, L., F. Oliviero, and R. Ramonda. Transforming growth factor-beta levels in synovial fluid of osteoarthritis with or without calcium pyrophosphate dihydrate crystals. J. Rheumatol. 30:420, 2003; (author reply 420-421).PubMedGoogle Scholar
  30. 30.
    Pupaibool, J., E. J. Fulnecky, R. L. Swords, Jr, W. W. Sistrunk, and A. D. Haddow. Alpha-defensin-novel synovial fluid biomarker for the diagnosis of periprosthetic joint infection. Int. Orthop. 40:2447–2452, 2016.CrossRefPubMedGoogle Scholar
  31. 31.
    Robson, H., T. Siebler, D. A. Stevens, S. M. Shalet, and G. R. Williams. Thyroid hormone acts directly on growth plate chondrocytes to promote hypertrophic differentiation and inhibit clonal expansion and cell proliferation. Endocrinology 141:3887–3897, 2000.CrossRefPubMedGoogle Scholar
  32. 32.
    Rohner, E., G. Matziolis, C. Perka, B. Fuchtmeier, T. Gaber, G. R. Burmester, F. Buttgereit, and P. Hoff. Inflammatory synovial fluid microenvironment drives primary human chondrocytes to actively take part in inflammatory joint diseases. Immunol. Res. 52:169–175, 2012.CrossRefPubMedGoogle Scholar
  33. 33.
    Sakai, T., F. Kambe, H. Mitsuyama, N. Ishiguro, K. Kurokouchi, M. Takigawa, H. Iwata, and H. Seo. Tumor necrosis factor alpha induces expression of genes for matrix degradation in human chondrocyte-like HCS-2/8 cells through activation of NF-kappaB: abrogation of the tumor necrosis factor alpha effect by proteasome inhibitors. J. Bone Miner. Res. 16:1272–1280, 2001.CrossRefPubMedGoogle Scholar
  34. 34.
    Sasaki, K., T. Hattori, T. Fujisawa, K. Takahashi, H. Inoue, and M. Takigawa. Nitric oxide mediates interleukin-1-induced gene expression of matrix metalloproteinases and basic fibroblast growth factor in cultured rabbit articular chondrocytes. J. Biochem. 123:431–439, 1998.CrossRefPubMedGoogle Scholar
  35. 35.
    Sato, E., T. Ando, J. Ichikawa, G. Okita, N. Sato, M. Wako, T. Ohba, S. Ochiai, T. Hagino, R. Jacobson, and H. Haro. High molecular weight hyaluronic acid increases the differentiation potential of the murine chondrocytic ATDC5 cell line. J. Orthop. Res. 32:1619–1627, 2014.CrossRefPubMedGoogle Scholar
  36. 36.
    Schmidt, T. A., and R. L. Sah. Effect of synovial fluid on boundary lubrication of articular cartilage. Osteoarthr. Cartil. 15:35–47, 2007.CrossRefPubMedGoogle Scholar
  37. 37.
    Schuerwegh, A. J., E. J. Dombrecht, W. J. Stevens, J. F. Van Offel, M. M. Kockx, C. H. Bridts, and L. S. De Clerck. Synovial fluid and peripheral blood immune complexes of patients with rheumatoid arthritis induce apoptosis in cytokine-activated chondrocytes. Rheumatol. Int. 27:901–909, 2007.CrossRefPubMedGoogle Scholar
  38. 38.
    Schwarz, I. M., and B. A. Hills. Surface-active phospholipid as the lubricating component of lubricin. Br. J. Rheumatol. 37:21–26, 1998.CrossRefPubMedGoogle Scholar
  39. 39.
    Shinoda, C., and S. Takaku. Interleukin-1 beta, interleukin-6, and tissue inhibitor of metalloproteinase-1 in the synovial fluid of the temporomandibular joint with respect to cartilage destruction. Oral Dis. 6:383–390, 2000.CrossRefPubMedGoogle Scholar
  40. 40.
    Shuler, F. D., H. I. Georgescu, C. Niyibizi, R. K. Studer, Z. Mi, B. Johnstone, R. D. Robbins, and C. H. Evans. Increased matrix synthesis following adenoviral transfer of a transforming growth factor beta1 gene into articular chondrocytes. J. Orthop. Res. 18:585–592, 2000.CrossRefPubMedGoogle Scholar
  41. 41.
    Snelling, S. J., S. Bas, G. J. Puskas, S. G. Dakin, D. Suva, A. Finckh, C. Gabay, P. Hoffmeyer, A. J. Carr, and A. Lubbeke. Presence of IL-17 in synovial fluid identifies a potential inflammatory osteoarthritic phenotype. PLoS ONE 12:e0175109, 2017.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Sun, Y., M. Lv, L. Zhou, V. Tam, F. Lv, D. Chan, H. Wang, Z. Zheng, K. M. Cheung, and V. Y. Leung. Enrichment of committed human nucleus pulposus cells expressing chondroitin sulfate proteoglycans under alginate encapsulation. Osteoarthr. Cartil. 23:1194–1203, 2015.CrossRefPubMedGoogle Scholar
  43. 43.
    Takada, K., J. Hirose, S. Yamabe, Y. Uehara, and H. Mizuta. Endoplasmic reticulum stress mediates nitric oxide-induced chondrocyte apoptosis. Biomed. Rep. 1:315–319, 2013.CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Tsuchida, A. I., M. Beekhuizen, M. Ct Hart, T. R. Radstake, W. J. Dhert, D. B. Saris, G. J. van Osch, and L. B. Creemers. Cytokine profiles in the joint depend on pathology, but are different between synovial fluid, cartilage tissue and cultured chondrocytes. Arthr. Res. Ther. 16:441, 2014.CrossRefGoogle Scholar
  45. 45.
    Verbruggen, A., L. S. De Clerck, C. H. Bridts, F. C. Breedveld, and W. J. Stevens. Influence of blood and synovial fluid immune complexes of patients with rheumatoid arthritis on production of nitric oxide and growth and viability of chondrocytes. J. Rheumatol. 27:35–40, 2000.PubMedGoogle Scholar
  46. 46.
    Yang, K. G., D. B. Saris, A. J. Verbout, L. B. Creemers, and W. J. Dhert. The effect of synovial fluid from injured knee joints on in vitro chondrogenesis. Tissue Eng. 12:2957–2964, 2006.CrossRefPubMedGoogle Scholar
  47. 47.
    Zhang, E., X. Yan, M. Zhang, X. Chang, Z. Bai, Y. He, and Z. Yuan. Aggrecanases in the human synovial fluid at different stages of osteoarthritis. Clin. Rheumatol. 32:797–803, 2013.CrossRefPubMedGoogle Scholar

Copyright information

© Biomedical Engineering Society 2018

Authors and Affiliations

  1. 1.Laboratory for Tissue Engineering and Morphogenesis, Department of Biomedical EngineeringRensselaer Polytechnic InstituteTroyUSA
  2. 2.Center for Biotechnology & Interdisciplinary StudiesRensselaer Polytechnic InstituteTroyUSA
  3. 3.Center for Modeling, Simulation and Imaging in MedicineRensselaer Polytechnic InstituteTroyUSA
  4. 4.Division of Orthopaedic SurgeryAlbany Medical CenterAlbanyUSA
  5. 5.Laboratory for Tissue Engineering and MorphogenesisRensselaer Polytechnic InstituteTroyUSA

Personalised recommendations