Annals of Biomedical Engineering

, Volume 40, Issue 9, pp 1874–1883 | Cite as

Fluid Flow-Induced Calcium Response in Early or Late Differentiated Osteoclasts

Article

Abstract

Intracellular calcium oscillation caused by receptor activator of nuclear factor kappa-B ligand has been demonstrated to promote the differentiation of osteoclasts. Osteoclasts are recruited on the surface of trabeculae, and are exposed to fluid flow caused by the deformation of the bone matrix. However, the roles of fluid shear stress (FSS) on calcium response during the differentiation process of osteoclasts are still unknown. In the current study, the formation of tartrate-resistant acid phosphatase-positive, multinucleated osteoclasts from RAW264.7 macrophage cells were induced by co-culturing them with the conditioned medium from MC3T3-E1 osteoblasts. The in situ observations showed a high correlation between the area and the nuclear number of osteoclasts. The cells were stimulated by FSS at different levels (1 or 10 dyne/cm2) before (0 day) or after being induced for 4 or 8 days. The mechanically-induced calcium response was recorded and analyzed. The results indicated a different property of calcium oscillation for the osteoclasts in different fusion stages (i.e., more calcium-responsive peaks appeared in small osteoclasts than those in the larger ones). The rates of calcium influx decreased and the time of recovery in osteoclast cytosol increased along with the fusion of osteoclasts. In addition, increasing the FSS level enhanced the calcium oscillation of osteoclasts at early induction (4 days). However, this effect was weakened at the late induction (8 days). The present work could help provide understanding regarding the mechanism of the involvement of calcium in mechanically induced bone remodeling.

Keywords

Osteoclasts Fluid shear stress Calcium oscillation Induction time Differentiation 

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China [30970707 (BH) and 31070829 (DZ)], the National Key Basic Research Foundation of China (2011CB710904), and the Knowledge Innovation Project of the CAS (KJCX2-YW-L08) (ML).

Conflict of interest

No conflict of interested is assigned to the manuscript.

References

  1. 1.
    Akchurin, T., T. Aissiou, et al. Complex dynamics of osteoclast formation and death in long-term cultures. PLoS ONE 3(5):11, 2008.CrossRefGoogle Scholar
  2. 2.
    Arai, F., T. Miyamoto, et al. Commitment and differentiation of osteoclast precursor cells by the sequential expression of c-Fms and receptor activator of nuclear factor kappaB (RANK) receptors. J. Exp. Med. 190(12):1741–1754, 1999.PubMedCrossRefGoogle Scholar
  3. 3.
    Dolmetsch, R. E., R. S. Lewis, et al. Differential activation of transcription factors induced by Ca2+ response amplitude and duration. Nature 386(6627):855–858, 1997.PubMedCrossRefGoogle Scholar
  4. 4.
    Dolmetsch, R. E., K. Xu, et al. Calcium oscillations increase the efficiency and specificity of gene expression. Nature 392(6679):933–936, 1998.PubMedCrossRefGoogle Scholar
  5. 5.
    Fahlgren, A., M. P. G. Bostrom, et al. Fluid pressure and flow as a cause of bone resorption. Acta Orthop. 81(4):508–516, 2010.PubMedCrossRefGoogle Scholar
  6. 6.
    Johansson, L., U. Edlund, et al. Bone resorption induced by fluid flow. J. Biomech. Eng. Trans. ASME 131(9):5, 2009.CrossRefGoogle Scholar
  7. 7.
    Jorgensen, N. R. Short-range intercellular calcium signaling in bone. APMIS Suppl 118:5–36, 2005.PubMedGoogle Scholar
  8. 8.
    Kurata, K., T. Uemura, et al. Mechanical strain effect on bone-resorbing activity and messenger RNA expressions of marker enzymes in isolated osteoclast culture. J. Bone Miner. Res. 16(4):722–730, 2001.PubMedCrossRefGoogle Scholar
  9. 9.
    Kuroda, Y., C. Hisatsune, et al. Osteoblasts induce Ca2+ oscillation-independent NFATc1 activation during osteoclastogenesis. Proc. Natl. Acad. Sci. USA 105(25):8643–8648, 2008.PubMedCrossRefGoogle Scholar
  10. 10.
    Lacey, D. L., E. Timms, et al. Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell 93(2):165–176, 1998.PubMedCrossRefGoogle Scholar
  11. 11.
    Liu, Y., L. Li, et al. Effects of fluid shear stress on bone resorption in rat osteoclasts. J. Biomed. Eng. 24(3):544–548, 2007.Google Scholar
  12. 12.
    Masuyama, R., J. Vriens, et al. TRPV4-mediated calcium influx regulates terminal differentiation of osteoclasts. Cell Metab. 8(3):257–265, 2008.PubMedCrossRefGoogle Scholar
  13. 13.
    Negishi-Koga, T., and H. Takayanagi. Ca2+-NFATc1 signaling is an essential axis of osteoclast differentiation. Immunol. Rev. 231:241–256, 2009.PubMedCrossRefGoogle Scholar
  14. 14.
    Rubin, J., X. Fan, et al. Osteoclastogenesis is repressed by mechanical strain in an in vitro model. J. Orthop. Res. 17(5):639–645, 1999.PubMedCrossRefGoogle Scholar
  15. 15.
    Sakai, H., Y. Moriura, et al. Phospholipase C-dependent Ca2+-sensing pathways leading to endocytosis and inhibition of the plasma membrane vacuolar H+-ATPase in osteoclasts. Am. J. Physiol. Cell Physiol. 299(3):C570–C578, 2010.PubMedCrossRefGoogle Scholar
  16. 16.
    Sanuki, R., C. Shionome, et al. Compressive force induces osteoclast differentiation via prostaglandin E(2) production in MC3T3-E1 cells. Connect. Tissue Res. 51(2):150–158, 2010.PubMedCrossRefGoogle Scholar
  17. 17.
    Tsuzuki, T., K. Okabe, et al. Osmotic membrane stretch increases cytosolic Ca(2+) and inhibits bone resorption activity in rat osteoclasts. Jpn. J. Physiol. 50(1):67–76, 2000.PubMedCrossRefGoogle Scholar
  18. 18.
    Wiebe, S. H., S. M. Sims, et al. Calcium signalling via multiple P2 purinoceptor subtypes in rat osteoclasts. Cell. Physiol. Biochem. 9(6):323–337, 1999.PubMedCrossRefGoogle Scholar
  19. 19.
    Xia, S. L., and J. Ferrier. Calcium signal induced by mechanical perturbation of osteoclasts. J. Cell. Physiol. 163(3):493–501, 1995.PubMedCrossRefGoogle Scholar
  20. 20.
    Xia, S. L., and J. Ferrier. Localized calcium signaling in multinucleated osteoclasts. J. Cell. Physiol. 167(1):148–155, 1996.PubMedCrossRefGoogle Scholar

Copyright information

© Biomedical Engineering Society 2012

Authors and Affiliations

  1. 1.Department of Stomatology, Faculty of Surgery, Peking Union Medical College HospitalChinese Academy of Medical SciencesBeijingPeople’s Republic of China
  2. 2.Key Laboratory of Microgravity, Center for Biomechanics and BioengineeringInstitute of Mechanics, Chinese Academy of SciencesBeijingPeople’s Republic of China

Personalised recommendations