Diffuse microdamage in bone activates anabolic response by osteoblasts via involvement of voltage-gated calcium channels

  • Hyungjin Jung
  • Ozan AkkusEmail author
Original Article



Matrix damage sustained by bone tissue is repaired by the concerted action of bone cells. Previous studies have reported extracellular calcium ([Ca2+]E) efflux to originate from regions of bone undergoing diffuse microdamage termed as “diffuse microdamage-induced calcium efflux” (DMICE). DMICE has also been shown to activate and increase intracellular calcium ([Ca2+]I) signaling in osteoblasts via the involvement of voltage-gated calcium channels (VGCC). Past studies have assessed early stage (< 1 h) responses of osteoblasts to DMICE. The current study tested the hypothesis that DMICE has longer-term sustained effect such that it induces anabolic response of osteoblasts.

Materials and methods

Osteoblasts derived from mouse calvariae were seeded on devitalized bovine bone wafers. Localized diffuse damage was induced in the vicinity of cells by bending. The response of osteoblasts to DMICE was evaluated by testing gene expression, protein synthesis and mineralized nodule formation.


Cells on damaged bone wafers showed a significant increase in RUNX2 and Osterix expression compared to non-loaded control. Also, RUNX2 and Osterix expression were suppressed significantly when the cells were treated with bepridil, a non-selective VGCC inhibitor, prior to loading. Significantly higher amounts of osteocalcin and mineralized nodules were synthesized by osteoblasts on diffuse damaged bone wafers, while bepridil treatment resulted in a significant decrease in osteocalcin production and mineralized nodule formation.


In conclusion, this study demonstrated that DMICE activates anabolic responses of osteoblasts through activation of VGCC. Future studies of osteoblast response to DMICE in vivo will help to clarify how bone cells repair diffuse microdamage.


Diffuse microdamage Calcium efflux Primary osteoblast Bone microdamage repair Voltage-gated calcium channel 



This study was funded by the National Science Foundation (NSF CMMI-1233413).

Compliance with ethical standards

Conflict of interest

The authors have declared that there is no conflict of interest.

Ethical approval

Animals were used in accordance with protocol (2014-0099) approved by the Institutional Animal Care and Use Committee (IACUC) of Case Western Reserve University. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.


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Copyright information

© The Japanese Society Bone and Mineral Research and Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Mechanical and Aerospace EngineeringCase Western Reserve UniversityClevelandUSA
  2. 2.Department of Biomedical EngineeringCase Western Reserve UniversityClevelandUSA
  3. 3.Department of OrthopedicsCase Western Reserve UniversityClevelandUSA

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