Skip to main content

Advertisement

SpringerLink
Log in

RNA Interference Targeting p110β Reduces Tumor Necrosis Factor-Alpha Production in Cellular Response to Wear Particles In vitro and Osteolysis In vivo

  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

Aseptic joint loosening is a key factor that reduces the life span of arthroplasty. There are currently few effective treatments for joint loosening except surgical revision. We explored the inhibitory effects of p110β-targeted small interfering RNA (siRNA) and lentivirus on particle-induced inflammatory cytokine expression in the murine macrophage cell line, RAW264.7. siRNA- and lentivirus-targeting p110β were transfected and infected prior to particle stimulation, respectively. Ceramic and titanium particles of different sizes were prepared to stimulate macrophages. Fluorescence microscopy showed that the efficiency of siRNA transfection and lentivirus infection were 74.2 ± 4.2 and 92.3 ± 2.6 %, respectively. TNF-alpha mRNA in the particle stimulation plus RNA interference (RNAi) groups were significantly lower compared with the particle stimulation-only groups (P < 0.05). Similarly, protein levels of TNF-alpha in RNAi-treated groups were significantly decreased after transfection or infection (P < 0.05). It showed that Phosphor-AKT (Ser473) activation was significantly reduced by RNAi through western blot. As assessed by CT, micro-CT and histological analysis, particle implantation induced a significant osteolysis in mice calvaria, which was limited by p110β lentivirus addition. These results suggested that p110β subtype of PI3K, followed by activation of Ser473, may possibly participate in the regulation of macrophages activity by wear particles, ultimately resulting in the TNF-α secretion and osteolysis.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  1. Wiklund, I., and B. Romanus. 1991. A comparison of quality of life before and after arthroplasty in patients who had arthrosis of the hip joint. The Journal of Bone and Joint Surgery American Volume 73(5): 765–769.

    PubMed  CAS  Google Scholar 

  2. Amstutz, H.C., P. Campbell, N. Kossovsky, and I.C. Clarke. 1992. Mechanism and clinical significance of wear debris-induced osteolysis. Clinical Orthopaedics and Related Research 276: 7–18.

    PubMed  Google Scholar 

  3. Charnley, J. 1975. Fracture of femoral prostheses in total hip replacement. A clinical study. Clinical Orthopaedics and Related Research 111: 105–120.

    Article  PubMed  Google Scholar 

  4. Akisue, T., T.W. Bauer, C.F. Farver, and Y. Mochida. 2002. The effect of particle wear debris on NFkappaB activation and pro-inflammatory cytokine release in differentiated THP-1 cells. Journal of Biomedical Materials Research 59(3): 507–515.

    Article  PubMed  CAS  Google Scholar 

  5. Qin, C.Q., Y. Ding, D.S. Huang, J. Xu, R.F. Ma, and J.B. Huang. 2011. Down-regulation of TNF-alpha by small interfering RNA inhibits particle-induced inflammation in vitro. Artificial Organs 35(7): 706–714.

    Article  PubMed  CAS  Google Scholar 

  6. Zysk, S.P., H.H. Gebhard, T. Kalteis, M. Schmitt-Sody, V. Jansson, K. Messmer, and A. Veihelmann. 2005. Particles of all sizes provoke inflammatory responses in vivo. Clinical Orthopaedics and Related Research 433: 258–264.

    Article  PubMed  Google Scholar 

  7. Puri, K.D., T.A. Doggett, C.Y. Huang, J. Douangpanya, J.S. Hayflick, M. Turner, J. Penninger, and T.G. Diacovo. 2005. The role of endothelial PI3Kgamma activity in neutrophil trafficking. Blood 106(1): 150–157.

    Article  PubMed  CAS  Google Scholar 

  8. Munugalavadla, V., J. Borneo, D.A. Ingram, and R. Kapur. 2005. p85alpha subunit of class IA PI-3 kinase is crucial for macrophage growth and migration. Blood 106(1): 103–109.

    Article  PubMed  CAS  Google Scholar 

  9. Huang, J.B., Y. Ding, D.S. Huang, A.J. Liang, W.K. Zeng, Z.P. Zeng, et al. 2013. Inhibition of the PI3K/AKT pathway reduces tumor necrosis factor-alpha production in the cellular response to wear particles in vitro. Artificial Organs 37(3): 298–307.

    Article  PubMed  CAS  Google Scholar 

  10. Boyce, B.F., P. Li, Z. Yao, Q. Zhang, I.R. Badell, E.M. Schwarz, R.J. O’Keefe, and L. Xing. 2005. TNF-alpha and pathologic bone resorption. The Keio Journal of Medicine 54(3): 127–131.

    Article  PubMed  CAS  Google Scholar 

  11. Gharbi, S.I., M.J. Zvelebil, S.J. Shuttleworth, T. Hancox, N. Saghir, J.F. Timms, and M.D. Waterfield. 2007. Exploring the specificity of the PI3K family inhibitor LY294002. Biochemical Journal 404(1): 15–21.

    Article  PubMed  CAS  Google Scholar 

  12. Hazeki, K., K. Nigorikawa, and O. Hazeki. 2007. Role of phosphoinositide 3-kinase in innate immunity. Biological and Pharmaceutical Bulletin 30(9): 1617–1623.

    Article  PubMed  CAS  Google Scholar 

  13. Fire, A., S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver, and C.C. Mello. 1998. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391(6669): 806–811.

    Article  PubMed  CAS  Google Scholar 

  14. Yang, M., and J. Mattes. 2008. Discovery, biology and therapeutic potential of RNA interference, microRNA and antagomirs. Pharmacology and Therapeutics 117(1): 94–104.

    Article  PubMed  CAS  Google Scholar 

  15. Salmon, P., and D. Trono. 2002. Lentiviral vectors for the gene therapy of lympho-hematological disorders. Current Topics in Microbiology and Immunology 261: 211–227.

    Article  PubMed  CAS  Google Scholar 

  16. von Knoch, M., D.E. Jewison, J.D. Sibonga, C. Sprecher, B.F. Morrey, F. Loer, D.J. Berry, and S.P. Scully. 2004. The effectiveness of polyethylene versus titanium particles in inducing osteolysis in vivo. Journal of Orthopaedic Research 22(2): 237–243.

    Article  Google Scholar 

  17. Greenfield, E.M., Y. Bi, A.A. Ragab, V.M. Goldberg, and R.R. Van De Motter. 2002. The role of osteoclast differentiation in aseptic loosening. Journal of Orthopaedic Research 20(1): 1–8.

    Article  PubMed  CAS  Google Scholar 

  18. Nanes, M.S. 2003. Tumor necrosis factor-alpha: molecular and cellular mechanisms in skeletal pathology. Gene 321: 1–15.

    Article  PubMed  CAS  Google Scholar 

  19. Tsukamoto, K., K. Hazeki, M. Hoshi, K. Nigorikawa, N. Inoue, T. Sasaki, and O. Hazeki. 2008. Critical roles of the p110 beta subtype of phosphoinositide 3-kinase in lipopolysaccharide-induced Akt activation and negative regulation of nitrite production in RAW 264.7 cells. Journal of Immunology 180(4): 2054–2061.

    CAS  Google Scholar 

  20. Smith, M.V., M.J. Lee, A.S. Islam, J.L. Rohrer, V.M. Goldberg, M.A. Beidelschies, and E.M. Greenfield. 2007. Inhibition of the PI3K-Akt signaling pathway reduces tumor necrosis factor-alpha production in response to titanium particles in vitro. The Journal of Bone and Joint Surgery. American Volume 89(5): 1019–1027.

    Article  PubMed  Google Scholar 

  21. Gartel, A.L., and E.S. Kandel. 2006. RNA interference in cancer. Biomolecular Engineering 23: 17–34.

    Article  PubMed  CAS  Google Scholar 

  22. Persengiev, S.P., X. Zhu, and M.R. Green. 2004. Nonspecific, concentration-dependent stimulation and repression of mammalian gene expression by small interfering RNAs (siRNAs). RNA 10(1): 12–18.

    Article  PubMed  CAS  Google Scholar 

  23. Lundstrom, K. 2003. Latest development in viral vectors for gene therapy. Trends in Biotechnology 21: 117–122.

    Article  PubMed  CAS  Google Scholar 

  24. Rovira, C., M.C. Guida, and A. Cayota. 2010. MicroRNAs and other small silencing RNAs in cancer. IUBMB Life 62(12): 859–868.

    Article  PubMed  CAS  Google Scholar 

  25. Couto, L.B., and K.A. High. 2010. Viral vector-mediated RNA interference. Current Opinion in Pharmacology 10(5): 534–542.

    Article  PubMed  CAS  Google Scholar 

  26. Hokaiwado, N., F. Takeshita, A. Banas, and T. Ochiya. 2008. RNAi-based drug discovery and its application to therapeutics. IDrugs 11(4): 274–278.

    PubMed  Google Scholar 

  27. Harper, S.Q., and P. Gonzalez-Alegre. 2008. Lentivirus-mediated RNA interference in mammalian neurons. Methods in Molecular Biology 442: 95–112.

    Article  PubMed  CAS  Google Scholar 

  28. Froelich, S., A. Tai, and P. Wang. 2010. Lentiviral vectors for immune cells targeting. Immunopharmacology and Immunotoxicology 32: 208–218.

    Article  PubMed  CAS  Google Scholar 

  29. Tsutsumi, R., C. Hock, C.D. Bechtold, S.T. Proulx, S.V. Bukata, H. Ito, H.A. Awad, T. Nakamura, R.J. O’Keefe, and E.M. Schwarz. 2008. Differential effects of biologic versus bisphosphonate inhibition of wear debris-induced osteolysis assessed by longitudinal micro-CT. Journal of Orthopaedic Research 26(10): 1340–1346.

    Article  PubMed  Google Scholar 

  30. Wooley, P.H., and E.M. Schwarz. 2004. Aseptic loosening. Gene Therapy 11(4): 402–407.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by funding from the National Natural Science Foundation of China (no. 81272020), the Science and Technology Planning Project of Guangzhou City (12S462060066), the Nature Science Foundation of Guangdong Province (S2012010009493), and Fundamental Research Funds for the Central Universities (10YKPY22).

Author information

Authors and Affiliations

  1. Department of Orthopaedic Surgery, Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University, 107 Yanjiangxi Road, 510120, Guangzhou, China

    Jian-bin Huang, Yue Ding, Dong-sheng Huang, Zhi-ping Guan & Mao-lin Zhang

  2. Department of Radiology, Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University, 107 Yanjiangxi Road, 510120, Guangzhou, China

    Wei-ke Zeng

Authors
  1. Jian-bin Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yue Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dong-sheng Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wei-ke Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhi-ping Guan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mao-lin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yue Ding.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, Jb., Ding, Y., Huang, Ds. et al. RNA Interference Targeting p110β Reduces Tumor Necrosis Factor-Alpha Production in Cellular Response to Wear Particles In vitro and Osteolysis In vivo . Inflammation 36, 1041–1054 (2013). https://doi.org/10.1007/s10753-013-9636-9

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10753-013-9636-9

KEY WORDS

Search

Navigation