Advertisement

Comparison of the cytotoxic and inflammatory responses of titanium particles with different methods for endotoxin removal in RAW264.7 macrophages

  • Huifeng Ding
  • Zhenan ZhuEmail author
  • Tingting Tang
  • Degang Yu
  • Bo Yu
  • Kerong Dai
Article

Abstract

It is generally accepted that periprosthetic bone resorption is initiated through aseptic inflammation aggravated by wear particles that are generated from artificial joint. However, some studies have demonstrated that “endotoxin-free” wear particles are almost completely unable to stimulate the macrophage-mediated production of proinflammatory cytokines. Here, we compare the titanium particles with different methods of endotoxin removal. The results indicated that different titanium particle preparation dosages did not significantly change particle size, morphology, and chemical composition. But it could cause variations in the endotoxin concentration of titanium particles and inflammatory responses in RAW264.7 macrophages. The particles with higher endotoxin levels correlated with more extensive inflammatory responses. When testing endotoxins using the supernatant of particle suspensions, it would lead to false negative results compared with testing the particle themselves. And when using the particles themselves, all the particles should be removed by centrifugation to avoid particle interference before the absorbance value was determined. Therefore, we suggest that research concerning wear particles should completely describe the endotoxin testing process, including endotoxin removal from particles and the details of endotoxin testing. Moreover, future research should focus on the surface of wear particles (the potential role of adherent endotoxin) rather than the particles themselves.

Keywords

Aseptic Loosening Wear Particle Titanium Particle Proinflammatory Cytokine Production Limulus Amebocyte Lysate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program) (Grant No. 2006AA02A137), Program for the Shanghai Key Laboratory of Orthopaedic Implant (Grant No. 08DZ2230300) and the National Natural Science Foundation of China (Grant No. 81001529). We also thank Prof. Jiake Xu from the University of Western Australia for his donation of the RAW264.7 cells that were stably transfected with a luciferase reporter gene.

References

  1. 1.
    Gallo J, Kaminek P, Ticha V, Rihakova P, Ditmar R. Particle disease. A comprehensive theory of periprosthetic osteolysis: a review. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2002;146(2):21–8.Google Scholar
  2. 2.
    Kolundzic R, Orlic D. Particle disease–aseptic loosening of the total hip endoprosthesis. Lijec Vjesn. 2008;130(1–2):16–20.Google Scholar
  3. 3.
    Purdue PE, Koulouvaris P, Nestor BJ, Sculco TP. The central role of wear debris in periprosthetic osteolysis. HSS J. 2006;2(2):102–13.CrossRefGoogle Scholar
  4. 4.
    Bi Y, Seabold JM, Kaar SG, Ragab AA, Goldberg VM, Anderson JM, et al. Adherent endotoxin on orthopedic wear particles stimulates cytokine production and osteoclast differentiation. J Bone Miner Res. 2001;16(11):2082–91.CrossRefGoogle Scholar
  5. 5.
    Cho DR, Shanbhag AS, Hong C-Y, Baran GR, Goldring SR. The role of adsorbed endotoxin in particle-induced stimulation of cytokine release. J Orthop Res. 2002;20(4):704–13.CrossRefGoogle Scholar
  6. 6.
    Brooks RA, Wimhurst JA, Rushton N. Endotoxin contamination of particles produces misleading inflammatory cytokine responses from macrophages in vitro. J Bone Joint Surg Br. 2002;84(2):295–9.CrossRefGoogle Scholar
  7. 7.
    Smith RA, Hallab NJ. In vitro macrophage response to polyethylene and polycarbonate-urethane particles. J Biomed Mater Res A. 2010;93(1):347–55.Google Scholar
  8. 8.
    Greenfield EM, Bi Y, Ragab AA, Goldberg VM, Nalepka JL, Seabold JM. Does endotoxin contribute to aseptic loosening of orthopedic implants? J Biomed Mater Res B Appl Biomater. 2005;72(1):179–85.CrossRefGoogle Scholar
  9. 9.
    Nalepka JL, Lee MJ, Kraay MJ, Marcus RE, Goldberg VM, Chen X, et al. Lipopolysaccharide found in aseptic loosening of patients with inflammatory arthritis. Clin Orthop Relat Res. 2006;451:229–35.CrossRefGoogle Scholar
  10. 10.
    Magalhaes PO, Lopes AM, Mazzola PG, Rangel-Yagui C, Penna TCV, Pessoa A Jr. Methods of endotoxin removal from biological preparations: a review. J Pharm Pharm Sci. 2007;10(3):388–404.Google Scholar
  11. 11.
    Ren W, Yang S-Y, Fang H-W, Hsu S, Wooley PH. Distinct gene expression of receptor activator of nuclear factor-kappaB and rank ligand in the inflammatory response to variant morphologies of UHMWPE particles. Biomaterials. 2003;24(26):4819–26.CrossRefGoogle Scholar
  12. 12.
    Fisher J, McEwen HMJ, Tipper JL, Galvin AL, Ingram J, Kamali A, et al. Wear, debris, and biologic activity of cross-linked polyethylene in the knee: benefits and potential concerns. Clin Orthop Relat Res. 2004;428:114–9.CrossRefGoogle Scholar
  13. 13.
    Ingram J, Matthews JB, Tipper J, Stone M, Fisher J, Ingham E. Comparison of the biological activity of grade GUR 1120 and GUR 415HP UHMWPE wear debris. Biomed Mater Eng. 2002;12(2):177–88.Google Scholar
  14. 14.
    St Pierre CA, Chan M, Iwakura Y, Ayers DC, Kurt-Jones EA, Finberg RW. Periprosthetic osteolysis: characterizing the innate immune response to titanium wear-particles. J Orthop Res. 2010;28(11):1418–24.Google Scholar
  15. 15.
    Trindade MC, Lind M, Sun D, Schurman DJ, Goodman SB, Smith RL. In vitro reaction to orthopaedic biomaterials by macrophages and lymphocytes isolated from patients undergoing revision surgery. Biomaterials. 2001;22(3):253–9.CrossRefGoogle Scholar
  16. 16.
    Kanaji A, Caicedo MS, Virdi AS, Sumner DR, Hallab NJ, Sena K. Co–Cr–Mo alloy particles induce tumor necrosis factor alpha production in MLO-Y4 osteocytes: a role for osteocytes in particle-induced inflammation. Bone. 2009;45(3):528–33.CrossRefGoogle Scholar
  17. 17.
    Yang S-Y, Yu H, Gong W, Wu B, Mayton L, Costello R, et al. Murine model of prosthesis failure for the long-term study of aseptic loosening. J Orthop Res. 2007;25(5):603–11.CrossRefGoogle Scholar
  18. 18.
    Zysk SP, Gebhard HH, Kalteis T, Schmitt-Sody M, Jansson V, Messmer K, et al. Particles of all sizes provoke inflammatory responses in vivo. Clin Orthop Relat Res. 2005;433:258–64.CrossRefGoogle Scholar
  19. 19.
    Utzschneider S, Becker F, Grupp TM, Sievers B, Paulus A, Gottschalk O, et al. Inflammatory response against different carbon fiber-reinforced PEEK wear particles compared with UHMWPE in vivo. Acta Biomater. 2010;6(11):4296–304.CrossRefGoogle Scholar
  20. 20.
    Kovacik MW, Mostardi RA, Neal DR, Bear TF, Askew MJ, Bender ET, et al. Differences in the surface composition of seemingly similar F75 cobalt-chromium micron-sized particulates can affect synovial fibroblast viability. Colloids Surf B Biointerfaces. 2008;65(2):269–75.CrossRefGoogle Scholar
  21. 21.
    Germain MA, Hatton A, Williams S, Matthews JB, Stone MH, Fisher J, et al. Comparison of the cytotoxicity of clinically relevant cobalt-chromium and alumina ceramic wear particles in vitro. Biomaterials. 2003;24(3):469–79.CrossRefGoogle Scholar
  22. 22.
    Ragab AA, Van De Motter R, Lavish SA, Goldberg VM, Ninomiya JT, Carlin CR, et al. Measurement and removal of adherent endotoxin from titanium particles and implant surfaces. J Orthop Res. 1999;17(6):803–9.CrossRefGoogle Scholar
  23. 23.
    von Knoch M, Jewison DE, Sibonga JD, Sprecher C, Morrey BF, Loer F, et al. The effectiveness of polyethylene versus titanium particles in inducing osteolysis in vivo. J Orthop Res. 2004;22(2):237–43.CrossRefGoogle Scholar
  24. 24.
    Lee S-S, Woo C-H, Chang J-D, Kim J-H. Roles of Rac and cytosolic phospholipase A2 in the intracellular signalling in response to titanium particles. Cell Signal. 2003;15(3):339–45.CrossRefGoogle Scholar
  25. 25.
    Gonzalez O, Smith RL, Goodman SB. Effect of size, concentration, surface area, and volume of polymethylmethacrylate particles on human macrophages in vitro. J Biomed Mater Res. 1996;30(4):463–73.CrossRefGoogle Scholar
  26. 26.
    Sterner T, Schutze N, Saxler G, Jakob F, Rader CP. Effects of clinically relevant alumina ceramic, zirconia ceramic and titanium particles of different sizes and concentrations on TNF-alpha release in a human macrophage cell line. Biomed Tech (Berl). 2004;49(12):340–4.CrossRefGoogle Scholar
  27. 27.
    Matthews JB, Besong AA, Green TR, Stone MH, Wroblewski BM, Fisher J, et al. Evaluation of the response of primary human peripheral blood mononuclear phagocytes to challenge with in vitro generated clinically relevant UHMWPE particles of known size and dose. J Biomed Mater Res. 2000;52(2):296–307.CrossRefGoogle Scholar
  28. 28.
    Choi MG, Koh HS, Kluess D, O’Connor D, Mathur A, Truskey GA, et al. Effects of titanium particle size on osteoblast functions in vitro and in vivo. Proc Natl Acad Sci USA. 2005;102(12):4578–83.CrossRefGoogle Scholar
  29. 29.
    Sommer B, Felix R, Sprecher C, Leunig M, Ganz R, Hofstetter W. Wear particles and surface topographies are modulators of osteoclastogenesis in vitro. J Biomed Mater Res A. 2005;72(1):67–76.CrossRefGoogle Scholar
  30. 30.
    Illgen RL, Forsythe TM, Pike JW, Laurent MP, Blanchard CR. Highly crosslinked vs conventional polyethylene particles—an in vitro comparison of biologic activities. J Arthroplasty. 2008;23(5):721–31.CrossRefGoogle Scholar
  31. 31.
    Kranz I, Gonzalez JB, Dorfel I, Gemeinert M, Griepentrog M, Klaffke D, et al. Biological response to micron- and nanometer-sized particles known as potential wear products from artificial hip joints: part II: Reaction of murine macrophages to corundum particles of different size distributions. J Biomed Mater Res A. 2009;89(2):390–401.Google Scholar
  32. 32.
    Geng DC, Xu YZ, Yang HL, Zhu XS, Zhu GM, Wang XB. Inhibition of titanium particle-induced inflammatory osteolysis through inactivation of cannabinoid receptor 2 by AM630. J Biomed Mater Res A. 2010;95(1):321–6.Google Scholar
  33. 33.
    Nakashima Y, Sun DH, Trindade MC, Maloney WJ, Goodman SB, Schurman DJ, et al. Signaling pathways for tumor necrosis factor-alpha and interleukin-6 expression in human macrophages exposed to titanium-alloy particulate debris in vitro. J Bone Joint Surg Am. 1999;81(5):603–15.Google Scholar
  34. 34.
    Fritz EA, Jacobs JJ, Glant TT, Roebuck KA. Chemokine IL-8 induction by particulate wear debris in osteoblasts is mediated by NF-kappaB. J Orthop Res. 2005;23(6):1249–57.Google Scholar
  35. 35.
    Wei X, Zhang X, Flick LM, Drissi H, Schwarz EM, O’Keefe RJ. Titanium particles stimulate COX-2 expression in synovial fibroblasts through an oxidative stress-induced, calpain-dependent, NF-kappaB pathway. Am J Physiol Cell Physiol. 2009;297(2):C310–20.CrossRefGoogle Scholar
  36. 36.
    Akisue T, Bauer TW, Farver CF, Mochida Y. The effect of particle wear debris on NFkappaB activation and pro-inflammatory cytokine release in differentiated THP-1 cells. J Biomed Mater Res. 2002;59(3):507–15.CrossRefGoogle Scholar
  37. 37.
    Wang C, Steer JH, Joyce DA, Yip KHM, Zheng MH, Xu J. 12-O-tetradecanoylphorbol-13-acetate (TPA) inhibits osteoclastogenesis by suppressing RANKL-induced NF-kappaB activation. J Bone Miner Res. 2003;18(12):2159–68.CrossRefGoogle Scholar
  38. 38.
    Hiromoto S, Hanawa T, Asami K. Composition of surface oxide film of titanium with culturing murine fibroblasts L929. Biomaterials. 2004;25(6):979–86.CrossRefGoogle Scholar
  39. 39.
    Mostardi RA, Kovacik MW, Ramsier RD, Bender ET, Finefrock JM, Bear TF, et al. A comparison of the effects of prosthetic and commercially pure metals on retrieved human fibroblasts: the role of surface elemental composition. Acta Biomater. 2010;6(2):702–7.CrossRefGoogle Scholar
  40. 40.
    Ingram JH, Stone M, Fisher J, Ingham E. The influence of molecular weight, crosslinking and counterface roughness on TNF-alpha production by macrophages in response to ultra high molecular weight polyethylene particles. Biomaterials. 2004;25(17):3511–22.CrossRefGoogle Scholar
  41. 41.
    Hallab NJ, Anderson S, Caicedo M, Brasher A, Mikecz K, Jacobs JJ. Effects of soluble metals on human peri-implant cells. J Biomed Mater Res A. 2005;74(1):124–40.Google Scholar
  42. 42.
    Yamamoto A, Honma R, Sumita M, Hanawa T. Cytotoxicity evaluation of ceramic particles of different sizes and shapes. J Biomed Mater Res A. 2004;68(2):244–56.CrossRefGoogle Scholar
  43. 43.
    Yamamoto Y, He P, Klein TW, Friedman H. Endotoxin induced cytotoxicity of macrophages is due to apoptosis caused by nitric oxide production. Innate Immun. 1994;1(3):181–7.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Huifeng Ding
    • 1
  • Zhenan Zhu
    • 1
    Email author
  • Tingting Tang
    • 1
  • Degang Yu
    • 1
  • Bo Yu
    • 1
  • Kerong Dai
    • 1
  1. 1.Department of Orthopaedic Surgery, Shanghai 9th People’s HospitalSchool of Medicine, Shanghai Jiao Tong UniversityShanghaiPeople’s Republic of China

Personalised recommendations