Skip to main content
SpringerLink
Log in

Full-Length Recombinant Human Proteoglycan 4 Interacts with Hyaluronan to Provide Cartilage Boundary Lubrication

  • Published:
Annals of Biomedical Engineering Aims and scope Submit manuscript

Abstract

Proteoglycan 4 (PRG4) is a mucin-like glycoprotein present in synovial fluid and at the surface of articular cartilage. The objectives of this study were to (1) assess the articular cartilage surface adsorption and in vitro cartilage boundary lubricating ability of full-length recombinant human PRG4 (rhPRG4), and (2) cartilage boundary lubricating ability of purified rhPRG4, both alone and in combination with hyaluronan (HA). rhPRG4 adsorption onto articular cartilage explants was assessed by immunohistochemistry and dot blot. An in vitro cartilage–cartilage friction test was used to assess rhPRG4’s cartilage boundary lubricating ability compared to bovine PRG4, and that of purified rhPRG4 both alone and in combination with HA. rhPRG4 was able to adsorb to the articular surface, as well as the cut surface, of cartilage explants. The kinetic coefficient of friction of rhPRG4 was similar to that of PRG4 (p = 0.16) and lower than phosphate-buffered saline (p < 0.05), while that of purified rhPRG4 + HA was significantly lower than rhPRG4 alone (p < 0.05). This study demonstrates that rhPRG4 can adsorb to an intact articular cartilage surface and functions as an effective boundary lubricant, both alone and with HA, and provides the foundation for in vivo evaluation of this clinically relevant full-length rhPRG4 for treatment of osteoarthritis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Al-Sharif, A., M. Jamal, L. Zhang, K. Larson, T. Schmidt, G. Jay, and K. Elsaid. Lubricin/proteoglycan 4 binding to CD44 receptor: a mechanism of lubricin’s suppression of pro-inflammatory cytokine induced synoviocyte proliferation. Arthritis Rheum. 67(6):1503–1513, 2015.

    Article  CAS  Google Scholar 

  2. Antonacci, J. M., T. A. Schmidt, L. A. Serventi, M. Z. Cai, Y. L. Shu, B. L. Schumacher, C. W. McIlwraith, and R. L. Sah. Effects of equine joint injury on boundary lubrication of articular cartilage by synovial fluid: role of hyaluronan. Arthritis Rheum. 64:2917–2926, 2012.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Ateshian, G. A. The role of interstitial fluid pressurization in articular cartilage lubrication. J. Biomech. 42:1163–1176, 2009.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Benz, M., N. Chen, G. Jay, and J. Israelachvili. Static forces, structure and flow properties of complex fluids in highly confined geometries. Ann. Biomed. Eng. 33:39–51, 2005.

    Article  PubMed  Google Scholar 

  5. Blain, E. J., S. J. Gilbert, R. J. Wardale, S. J. Capper, D. J. Mason, and V. C. Duance. Up-regulation of matrix metalloproteinase expression and activation following cyclical compressive loading of articular cartilage in vitro. Arch. Biochem. Biophys. 396:49–55, 2001.

    Article  CAS  PubMed  Google Scholar 

  6. Bonnevie, E. D., C. Secchieri, D. Galesso and L. J. Bonassar. Stribeck analysis of synovial lubricants: lubricating mechanisms and interaction of hyaluronic acid and lubricin. Trans. Orthop. Res. Soc. Poster No: 0437, 2014.

  7. Chang, D. P., N. I. Abu-Lail, J. M. Coles, F. Guilak, G. D. Jay, and S. Zauscher. Friction force microscopy of lubricin and hyaluronic acid between hydrophobic and hydrophilic surfaces. Soft Matter 5:3438–3445, 2009.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Corvelli M., B. Che, C. Saeui, A. Singh, and J. Elisseeff. Biodynamic performance of hyaluronic acid versus synovial fluid of the knee in osteoarthritis. Methods 2015. doi:10.1016/j.ymeth.2015.03.019.

    PubMed  Google Scholar 

  9. Das, S., X. Banquy, B. Zappone, G. W. Greene, G. D. Jay, and J. N. Israelachvili. Synergistic interactions between grafted hyaluronic acid and lubricin provide enhanced wear protection and lubrication. Biomacromolecules 14:1669–1677, 2013.

    Article  CAS  PubMed  Google Scholar 

  10. Elsaid, K. A., B. C. Fleming, H. L. Oksendahl, J. T. Machan, P. D. Fadale, M. J. Hulstyn, R. Shalvoy, and G. D. Jay. Decreased lubricin concentrations and markers of joint inflammation in the synovial fluid of patients with anterior cruciate ligament injury. Arthritis Rheum. 58:1707–1715, 2008.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Elsaid, K. A., G. D. Jay, M. L. Warman, D. K. Rhee, and C. O. Chichester. Association of articular cartilage degradation and loss of boundary-lubricating ability of synovial fluid following injury and inflammatory arthritis. Arthritis Rheum. 52:1746–1755, 2005.

    Article  CAS  PubMed  Google Scholar 

  12. Elsaid, K. A., L. Zhang, Z. Shaman, C. Patel, T. A. Schmidt, and G. D. Jay. The impact of early intra-articular administration of interleukin-1 receptor antagonist on lubricin metabolism and cartilage degeneration in an anterior cruciate ligament transection model. Osteoarthr. Cartil. 23:114–121, 2015.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Elsaid, K. A., L. Zhang, K. Waller, J. Tofte, E. Teeple, B. C. Fleming, and G. D. Jay. The impact of forced joint exercise on lubricin biosynthesis from articular cartilage following ACL transection and intra-articular lubricin’s effect in exercised joints following ACL transection. Osteoarthr. Cartil. 20:940–948, 2012.

    Article  CAS  PubMed  Google Scholar 

  14. Flannery, C. R., R. Zollner, C. Corcoran, A. R. Jones, A. Root, M. A. Rivera-Bermudez, T. Blanchet, J. P. Gleghorn, L. J. Bonassar, A. M. Bendele, E. A. Morris, and S. S. Glasson. Prevention of cartilage degeneration in a rat model of osteoarthritis by intraarticular treatment with recombinant lubricin. Arthritis Rheum. 60:840–847, 2009.

    Article  CAS  PubMed  Google Scholar 

  15. Forster, H., and J. Fisher. The influence of loading time and lubricant on the friction of articular cartilage. Proc. Inst. Mech. Eng. H 210:109–119, 1996.

    Article  CAS  PubMed  Google Scholar 

  16. Gleghorn, J. P., A. R. Jones, C. R. Flannery, and L. J. Bonassar. Boundary mode lubrication of articular cartilage by recombinant human lubricin. J. Orthop. Res. 27:771–777, 2009.

    Article  CAS  PubMed  Google Scholar 

  17. Greene, G. W., X. Banquy, D. W. Lee, D. D. Lowrey, J. Yu, and J. N. Israelachvili. Adaptive mechanically controlled lubrication mechanism found in articular joints. Proc Natl Acad Sci USA 108:5255–5259, 2011.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Jay, G. D., K. A. Elsaid, K. A. Kelly, S. C. Anderson, L. Zhang, E. Teeple, K. Waller, and B. C. Fleming. Prevention of cartilage degeneration and gait asymmetry by lubricin tribosupplementation in the rat following anterior cruciate ligament transection. Arthritis Rheum. 64:1162–1171, 2012.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Jay, G. D., B. C. Fleming, B. A. Watkins, K. A. McHugh, S. C. Anderson, L. X. Zhang, E. Teeple, K. A. Waller, and K. A. Elsaid. Prevention of cartilage degeneration and restoration of chondroprotection by lubricin tribosupplementation in the rat following anterior cruciate ligament transection. Arthritis Rheum. 62:2382–2391, 2010.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Jay, G. D., J. R. Torres, D. K. Rhee, H. J. Helminen, M. M. Hytinnen, C. J. Cha, K. Elsaid, K. S. Kim, Y. Cui, and M. L. Warman. Association between friction and wear in diarthrodial joints lacking lubricin. Arthritis Rheum. 56:3662–3669, 2007.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Jay, G. D., J. R. Torres, M. L. Warman, M. C. Laderer, and K. S. Breuer. The role of lubricin in the mechanical behavior of synovial fluid. Proc Natl Acad Sci USA 104:6194–6199, 2007.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Jay, G. D., and K. A. Waller. The biology of Lubricin: near frictionless joint motion. Matrix Biol. 39:17–24, 2014.

    Article  CAS  PubMed  Google Scholar 

  23. Jones, A. R., J. P. Gleghorn, C. E. Hughes, L. J. Fitz, R. Zollner, S. D. Wainwright, B. Caterson, E. A. Morris, L. J. Bonassar, and C. R. Flannery. Binding and localization of recombinant lubricin to articular cartilage surfaces. J. Orthop. Res. 25:283–292, 2007.

    Article  CAS  PubMed  Google Scholar 

  24. Kwiecinski, J. J., S. G. Dorosz, T. E. Ludwig, S. Abubacker, M. K. Cowman, and T. A. Schmidt. The effect of molecular weight on hyaluronan’s cartilage boundary lubricating ability—alone and in combination with proteoglycan 4. Osteoarthr. Cartil. 19:1356–1362, 2011.

    Article  CAS  PubMed  Google Scholar 

  25. Lee, J. H., J. B. Fitzgerald, M. A. Dimicco, and A. J. Grodzinsky. Mechanical injury of cartilage explants causes specific time-dependent changes in chondrocyte gene expression. Arthritis Rheum. 52:2386–2395, 2005.

    Article  CAS  PubMed  Google Scholar 

  26. Nugent-Derfus, G. E., A. H. Chan, B. L. Schumacher, and R. L. Sah. PRG4 exchange between the articular cartilage surface and synovial fluid. J. Orthop. Res. 25:1269–1276, 2007.

    Article  CAS  PubMed  Google Scholar 

  27. Rhee, D. K., J. Marcelino, M. Baker, Y. Gong, P. Smits, V. Lefebvre, G. D. Jay, M. Stewart, H. Wang, M. L. Warman, and J. D. Carpten. The secreted glycoprotein lubricin protects cartilage surfaces and inhibits synovial cell overgrowth. J. Clin. Invest. 115:622–631, 2005.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Samsom, M. L., S. Morrison, N. Masala, B. D. Sullivan, D. A. Sullivan, H. Sheardown, and T. A. Schmidt. Characterization of full-length recombinant human proteoglycan 4 as an ocular surface boundary lubricant. Exp. Eye Res. 127:14–19, 2014.

    Article  CAS  PubMed  Google Scholar 

  29. Schmidt, T. A., N. S. Gastelum, E. H. Han, G. E. Nugent-Derfus, B. L. Schumacher, and R. L. Sah. Differential regulation of proteoglycan 4 metabolism in cartilage by IL-1alpha, IGF-I, and TGF-beta1. Osteoarthr. Cartil. 16:90–97, 2008.

    Article  CAS  PubMed  Google Scholar 

  30. Schmidt, T. A., N. S. Gastelum, Q. T. Nguyen, B. L. Schumacher, and R. L. Sah. Boundary lubrication of articular cartilage: role of synovial fluid constituents. Arthritis Rheum. 56:882–891, 2007.

    Article  CAS  PubMed  Google Scholar 

  31. Schmidt, T. A., A. H. Plaas, and J. D. Sandy. Disulfide-bonded multimers of proteoglycan 4 (PRG4) are present in normal synovial fluids. Biochim. Biophys. Acta 1790:375–384, 2009.

    Article  CAS  PubMed  Google Scholar 

  32. Schmidt, T. A., and R. L. Sah. Effect of synovial fluid on boundary lubrication of articular cartilage. Osteoarthr. Cartil. 15:35–47, 2007.

    Article  CAS  PubMed  Google Scholar 

  33. Schumacher, B. L., T. A. Schmidt, M. S. Voegtline, A. C. Chen, and R. L. Sah. Proteoglycan 4 (PRG4) synthesis and immunolocalization in bovine meniscus. J. Orthop. Res. 23:562–568, 2005.

    Article  CAS  PubMed  Google Scholar 

  34. Seror, J., L. Zhu, R. Goldberg, A. J. Day, and J. Klein. Supramolecular synergy in the boundary lubrication of synovial joints. Nat. Commun. 6:6497, 2014.

    Article  Google Scholar 

  35. Singh, A., M. Corvelli, S. A. Unterman, K. A. Wepasnick, P. McDonnell, and J. H. Elisseeff. Enhanced lubrication on tissue and biomaterial surfaces through peptide-mediated binding of hyaluronic acid. Nat. Mater. 13:988–995, 2014.

    Article  CAS  PubMed  Google Scholar 

  36. Steele, B. L., M. C. Alvarez-Veronesi, and T. A. Schmidt. Molecular weight characterization of PRG4 proteins using multi-angle laser light scattering (MALLS). Osteoarthr. Cartil. 21:498–504, 2013.

    Article  CAS  PubMed  Google Scholar 

  37. Swann, D. A., R. B. Hendren, E. L. Radin, S. L. Sotman, and E. A. Duda. The lubricating activity of synovial fluid glycoproteins. Arthritis Rheum. 24:22–30, 1981.

    Article  CAS  PubMed  Google Scholar 

  38. Teeple, E., K. A. Elsaid, G. D. Jay, L. Zhang, G. J. Badger, M. Akelman, T. F. Bliss, and B. C. Fleming. Effects of supplemental intra-articular lubricin and hyaluronic acid on the progression of posttraumatic arthritis in the anterior cruciate ligament-deficient rat knee. Am. J. Sports Med. 39:164–172, 2011.

    Article  PubMed  PubMed Central  Google Scholar 

  39. Waller, K. A., L. X. Zhang, K. A. Elsaid, B. C. Fleming, M. L. Warman, and G. D. Jay. Role of lubricin and boundary lubrication in the prevention of chondrocyte apoptosis. Proc. Natl. Acad. Sci. USA 110:5852–5857, 2013.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Watt, F. E., H. M. Ismail, A. Didangelos, M. Peirce, T. L. Vincent, R. Wait, and J. Saklatvala. Src and fibroblast growth factor 2 independently regulate signaling and gene expression induced by experimental injury to intact articular cartilage. Arthritis Rheum. 65:397–407, 2013.

    Article  CAS  PubMed  Google Scholar 

  41. Zappone, B., M. Ruths, G. W. Greene, G. D. Jay, and J. N. Israelachvili. Adsorption, lubrication, and wear of lubricin on model surfaces: polymer brush-like behavior of a glycoprotein. Biophys. J. 92:1693–1708, 2007.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We would like to thank the Dr. Umberto Banderalli for his help with the confocal microscopy and Dr. Tak Fung for his assistance with statistical data analysis.

Funding Source

This work was supported by funding from the Natural Sciences and Engineering Research Council of Canada (355591), The Arthritis Society, the Alberta Innovates Health Solutions Team in Osteoarthritis.

Conflict of interest

TAS and GDJ have a financial interest in, and are named inventors on issued patents held by a commercial entity (Lubris) developing rhPRG4 for therapeutic uses. TAS is also a paid consultant for the same entity.

Author information

Authors and Affiliations

  1. Biomedical Engineering Graduate Program, University of Calgary, 2500 University Dr NW, KNB 426, Calgary, AB, T2N 1N4, Canada

    Saleem Abubacker, Samuel G. Dorosz & Tannin A. Schmidt

  2. McCaig Institute of Bone and Joint Health, University of Calgary, Calgary, AB, Canada

    Saleem Abubacker, Samuel G. Dorosz, Dragana Ponjevic, John R. Matyas & Tannin A. Schmidt

  3. Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada

    Dragana Ponjevic & John R. Matyas

  4. Center for Biomedical Engineering and the School of Engineering, Brown University, Providence, RI, USA

    Gregory D. Jay

  5. Faculty of Kinesiology, University of Calgary, Calgary, AB, Canada

    Tannin A. Schmidt

Authors
  1. Saleem Abubacker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Samuel G. Dorosz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dragana Ponjevic
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gregory D. Jay
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. John R. Matyas
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tannin A. Schmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tannin A. Schmidt.

Additional information

Associate Editor Eric Darling oversaw the review of this article.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Abubacker, S., Dorosz, S.G., Ponjevic, D. et al. Full-Length Recombinant Human Proteoglycan 4 Interacts with Hyaluronan to Provide Cartilage Boundary Lubrication. Ann Biomed Eng 44, 1128–1137 (2016). https://doi.org/10.1007/s10439-015-1390-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10439-015-1390-8

Keywords

Search

Navigation