Skip to main content

P2X7 Nucleotide Receptor Plays an Important Role in Callus Remodeling During Fracture Repair

Calcified Tissue International volume 84, pages 405–412 (2009)Cite this article

Abstract

The P2X7 nucleotide receptor (P2X7R) is an ATP-gated ion channel expressed in bone cells. Homozygous null P2X7R (P2X7R–/–) mice have reduced bone formation, so we hypothesized that P2X7R–/– mice have impaired fracture healing compared to P2X7R+/+ control mice. To test the hypothesis, adult P2X7R–/– mice and P2X7R+/+ mice were studied. Osteotomy of the right femur was performed and a stainless-steel pin was inserted into the medullary cavity to stabilize the fracture site. No differences in callus development were seen in the radiograph, micro computed tomography, or dual-energy x-ray absorptiometry measurements. Mechanical testing showed that the recovery of ultimate force, stiffness, and energy to failure were slightly decreased in P2X7R–/– mice compared with the control. Histomorphometric measurements of the callus revealed that mineralizing surface and bone formation were significantly decreased, by 22% (p < 0.001) and 29% (p < 0.05), respectively, in P2X7R–/– mice in comparison with the wild-type control. These data show that a null mutation of the P2X7R does not affect the amount of callus formed in our osteotomy fracture model. However, callus remodeling was significantly delayed. Our data suggest the different role of the P2X7R in woven bone and lamellar bone formation.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Genetos DC, Geist DJ, Liu D, Donahue HJ, Duncan RL (2005) Fluid shear-induced ATP secretion mediates prostaglandin release in MC3T3-E1 osteoblasts. J Bone Miner Res 20:41–49

    PubMed  Article  CAS  Google Scholar 

  2. Li J, Liu D, Ke HZ, Duncan RL, Turner CH (2005) The P2X7 nucleotide receptor mediates skeletal mechanotransduction. J Biol Chem 280:42952–42959

    PubMed  Article  CAS  Google Scholar 

  3. Bowler WB, Dixon CJ, Halleux C, Maier R, Bilbe G, Fraser WD, Gallagher JA, Hipskind RA (1999) Signaling in human osteoblasts by extracellular nucleotides. Their weak induction of the c-fos proto-oncogene via Ca2+ mobilization is strongly potentiated by a parathyroid hormone/cAMP-dependent protein kinase pathway independently of mitogen-activated protein kinase. J Biol Chem 274:14315–14324

    PubMed  Article  CAS  Google Scholar 

  4. North RA (2002) Molecular physiology of P2X receptors. Physiol Rev 82:1013–1067

    PubMed  CAS  Google Scholar 

  5. Surprenant A, Rassendren F, Kawashima E, North RA, Buell G (1996) The cytolytic P2Z receptor for extracellular ATP identified as a P2X receptor (P2X7). Science 272:735–738

    PubMed  Article  CAS  Google Scholar 

  6. Ferrari D, Chiozzi P, Falzoni S, Dal Susino M, Melchiorri L, Baricordi OR, Di Virgilio F (1997) Extracellular ATP triggers IL-1 beta release by activating the purinergic P2Z receptor of human macrophages. J Immunol 159:1451–1458

    PubMed  CAS  Google Scholar 

  7. Ferrari D, Los M, Bauer MK, Vandenabeele P, Wesselborg S, Schulze-Osthoff K (1999) P2Z purinoreceptor ligation induces activation of caspases with distinct roles in apoptotic and necrotic alterations of cell death. FEBS Lett 447:71–75

    PubMed  Article  CAS  Google Scholar 

  8. Kim M, Jiang LH, Wilson HL, North RA, Surprenant A (2001) Proteomic and functional evidence for a P2X7 receptor signalling complex. EMBO J 20:6347–6358

    PubMed  Article  CAS  Google Scholar 

  9. Ke HZ, Qi H, Weidema AF, Zhang Q, Panupinthu N, Crawford DT, Grasser WA, Paralkar VM, Li M, Audoly LP, Gabel CA, Jee WS, Dixon SJ, Sims SM, Thompson DD (2003) Deletion of the P2X7 nucleotide receptor reveals its regulatory roles in bone formation and resorption. Mol Endocrinol 17:1356–1367

    PubMed  Article  CAS  Google Scholar 

  10. Cui L, Ma YF, Yao W, Zhou H, Setterberg RB, Liang TC, Jee WS (2001) Cancellous bone of aged rats maintains its capacity to respond vigorously to the anabolic effects of prostaglandin E2 by modeling-dependent bone gain. J Bone Miner Metab 19:29–37

    PubMed  Article  CAS  Google Scholar 

  11. Mo A, Yao W, Li C, Tian X, Su M, Ling Y, Zhang Q, Setterberg RB, Jee WS (2002) Bipedal stance exercise and prostaglandin E2 (PGE2) and its synergistic effect in increasing bone mass and in lowering the PGE2 dose required to prevent ovariectomized-induced cancellous bone loss in aged rats. Bone 31:402–406

    PubMed  Article  CAS  Google Scholar 

  12. Yao W, Jee WS, Zhou H, Lu J, Cui L, Setterberg R, Liang T, Ma Y (1999) Anabolic effect of prostaglandin E2 on cortical bone of aged male rats comes mainly from modeling-dependent bone gain. Bone 25:697–702

    PubMed  Article  CAS  Google Scholar 

  13. Solle M, Labasi J, Perregaux DG, Stam E, Petrushova N, Koller BH, Griffiths RJ, Gabel CA (2001) Altered cytokine production in mice lacking P2X(7) receptors. J Biol Chem 276:125–132

    PubMed  Article  CAS  Google Scholar 

  14. Bonnarens F, Einhorn TA (1984) Production of a standard closed fracture in laboratory animal bone. J Orthop Res 2:97–101

    PubMed  Article  CAS  Google Scholar 

  15. Li C, Mori S, Li J, Kaji Y, Akiyama T, Kawanishi J, Norimatsu H (2001) Long-term effect of incadronate disodium (YM-175) on fracture healing of femoral shaft in growing rats. J Bone Miner Res 16:429–436

    PubMed  Article  CAS  Google Scholar 

  16. Li J, Mori S, Kaji Y, Kawanishi J, Akiyama T, Norimatsu H (2000) Concentration of bisphosphonate (incadronate) in callus area and its effects on fracture healing in rats. J Bone Miner Res 15:2042–2051

    PubMed  Article  CAS  Google Scholar 

  17. Li J, Mori S, Kaji Y, Mashiba T, Kawanishi J, Norimatsu H (1999) Effect of bisphosphonate (incadronate) on fracture healing of long bones in rats. J Bone Miner Res 14:969–979

    PubMed  Article  CAS  Google Scholar 

  18. Dimitriou R, Tsiridis E, Giannoudis PV (2005) Current concepts of molecular aspects of bone healing. Injury 36:1392–1404

    PubMed  Article  Google Scholar 

  19. Gerstenfeld LC, Alkhiary YM, Krall EA, Nicholls FH, Stapleton SN, Fitch JL, Bauer M, Kayal R, Graves DT, Jepsen KJ, Einhorn TA (2006) Three-dimensional reconstruction of fracture callus morphogenesis. J Histochem Cytochem 54:1215–1228

    PubMed  Article  CAS  Google Scholar 

  20. Ai-Aql ZS, Alagl AS, Graves DT, Gerstenfeld LC, Einhorn TA (2008) Molecular mechanisms controlling bone formation during fracture healing and distraction osteogenesis. J Dent Res 87:107–118

    PubMed  Article  CAS  Google Scholar 

  21. Pelegrin P, Barroso-Gutierrez C, Surprenant A (2008) P2X7 receptor differentially couples to distinct release pathways for IL-1beta in mouse macrophage. J Immunol 180:7147–7157

    PubMed  CAS  Google Scholar 

  22. Shigemoto-Mogami Y, Koizumi S, Tsuda M, Ohsawa K, Kohsaka S, Inoue K (2001) Mechanisms underlying extracellular ATP-evoked interleukin-6 release in mouse microglial cell line, MG-5. J Neurochem 78:1339–1349

    PubMed  Article  CAS  Google Scholar 

  23. Wang CM, Chang YY, Sun SH (2003) Activation of P2X7 purinoceptor-stimulated TGF-beta 1 mRNA expression involves PKC/MAPK signalling pathway in a rat brain-derived type-2 astrocyte cell line, RBA-2. Cell Signal 15:1129–1137

    PubMed  Article  Google Scholar 

  24. Korcok J, Raimundo LN, Ke HZ, Sims SM, Dixon SJ (2004) Extracellular nucleotides act through P2X7 receptors to activate NF-kappaB in osteoclasts. J Bone Miner Res 19:642–651

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by National Institutes of Health Grant R01AR046530 (C.H.T.) and AR051901 (R.L.D.).

Author information

Affiliations

  1. Department of Biology, Indiana University Purdue University Indianapolis, 723 West Michigan Street, SL 306, Indianapolis, IN, 46202, USA

    Jiliang Li

  2. Department of Biomedical Engineering, Indiana University Purdue University Indianapolis, Indianapolis, IN, 46202, USA

    Rachel Meyer & Charles H. Turner

  3. Department of Biological Sciences, University of Delaware, Newark, DE, 19716, USA

    Randall L. Duncan

Authors
  1. Jiliang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rachel Meyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Randall L. Duncan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Charles H. Turner
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jiliang Li.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Li, J., Meyer, R., Duncan, R.L. et al. P2X7 Nucleotide Receptor Plays an Important Role in Callus Remodeling During Fracture Repair. Calcif Tissue Int 84, 405–412 (2009). https://doi.org/10.1007/s00223-009-9237-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00223-009-9237-7

Keywords

  • P2X7 receptor
  • Fracture repair
  • Bone remodeling
  • Bone density
  • Histomorphometry
  • Biomechanics