• Open Access

Shockwaves Induce Osteogenic Differentiation of Human Mesenchymal Stem Cells Through ATP Release and Activation of P2X7 Receptors§

Authors

  • Dahui Sun,

    1. Department of Orthopedics, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    Search for more papers by this author
  • Wolfgang G. Junger,

    1. Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA
    2. Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria
    Search for more papers by this author
  • Changji Yuan,

    1. Department of Hematology, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    Search for more papers by this author
  • Wenyan Zhang,

    1. Institute of Virology and AIDS research, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    Search for more papers by this author
  • Yi Bao,

    1. Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA
    Search for more papers by this author
  • Daming Qin,

    1. Department of Orthopedics, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    Search for more papers by this author
  • Chengxue Wang,

    1. Department of Orthopedics, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    Search for more papers by this author
  • Lei Tan,

    1. Department of Orthopedics, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    Search for more papers by this author
  • Baochang Qi,

    1. Department of Orthopedics, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    Search for more papers by this author
  • Dong Zhu,

    1. Department of Orthopedics, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    Search for more papers by this author
  • Xizheng Zhang,

    1. Institute of Medical Equipment Military Medical Science Academy of the PLA, Tian Jin, People's Republic of China
    Search for more papers by this author
  • Tiecheng Yu

    Corresponding author
    1. Department of Orthopedics, The First Norman Bethune Hospital of Jilin University, Changchun, People's Republic of China
    • Department of Orthopedics, First Norman Bethune Hospital of Jilin University, Changchun 130021, People's Republic of China
    Search for more papers by this author
    • Telephone: 86-431-85098981; Fax: 0086-431-85654528


  • Author contributions: D.S.: performed experiments and wrote the manuscript; W.G.J.: helped with study design and finalizing manuscript; C.Y.: provided administrative support; W.Z.: provided study material; Y.B.: prepared figures; D.Q. and C.W.: prepared samples; L.T. and B.Q.: performed experiments and collected data; D.Z. and X.Z.: provided administrative support; T.Y.: Developed concept, designed the study, provided overall guidance, and finalized manuscript.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    first published online in STEM CELLS EXPRESS February 13, 2013.

Abstract

Shockwave treatment promotes bone healing of nonunion fractures. In this study, we investigated whether this effect could be due to adenosine 5′-triphosphate (ATP) release-induced differentiation of human mesenchymal stem cells (hMSCs) into osteoprogenitor cells. Cultured bone marrow-derived hMSCs were subjected to shockwave treatment and ATP release was assessed. Osteogenic differentiation and mineralization of hMSCs were evaluated by examining alkaline phosphatase activity, osteocalcin production, and calcium nodule formation. Expression of P2X7 receptors and c-fos and c-jun mRNA was determined with real-time reverse transcription polymerase chain reaction and Western blotting. P2X7-siRNA, apyrase, P2 receptor antagonists, and p38 MAPK inhibitors were used to evaluate the roles of ATP release, P2X7 receptors, and p38 MAPK signaling in shockwave-induced osteogenic hMSCs differentiation. Shockwave treatment released significant amounts (∼7 μM) of ATP from hMSCs. Shockwaves and exogenous ATP induced c-fos and c-jun mRNA transcription, p38 MAPK activation, and hMSC differentiation. Removal of ATP with apyrase, targeting of P2X7 receptors with P2X7-siRNA or selective antagonists, or blockade of p38 MAPK with SB203580 prevented osteogenic differentiation of hMSCs. Our findings indicate that shockwaves release cellular ATP that activates P2X7 receptors and downstream signaling events that caused osteogenic differentiation of hMSCs. We conclude that shockwave therapy promotes bone healing through P2X7 receptor signaling, which contributes to hMSC differentiation. STEM Cells 2013;31:1170–1180

Ancillary