Ascorbic acid accelerates osteoclast formation and death

Bone. 2010 May;46(5):1336-43. doi: 10.1016/j.bone.2009.11.021. Epub 2009 Nov 20.


Ascorbic acid (AA) plays a key role in bone formation. However controversy remains about the effect of AA on cells responsible for bone destruction, osteoclasts. We investigated the effect of AA on osteoclastogenesis using primary mouse bone marrow cultures and monocytic RAW 264.7 cells treated with osteoclastogenic factors RANKL and MCSF. Treatment with AA resulted in significant increase in osteoclast number, size and nucleation. To assess osteoclast oxidative stress level, a ratio of reduced (GSH) to oxidized (GSSG) glutathione and the total glutathione content (GSH(t)) were evaluated. Osteoclast differentiation was associated with a decrease in GSH/GSSG and GSH(t). AA induced further decrease in both parameters, and resulted in significant production of H(2)O(2), indicating its pro-oxidant action. At low concentration, H(2)O(2) induced similar effects to AA, although less potently, and catalase partially inhibited AA-induced osteoclastogenesis. To assess the modification in osteoclast metabolism, the mitochondrial activity was evaluated using JC-1 and the ATP levels were assessed. Osteoclast formation was associated with the increase in mitochondrial activity and ATP concentration, which were further increased in the presence of AA. Importantly, the stimulatory effect of AA was only evident at early phase of osteoclastogenesis, whereas at the late stage AA significantly accelerated osteoclast death. Thus, during osteoclastogenesis AA acts as an oxidant, first stimulating osteoclast formation, but later limiting osteoclast lifespan. This duality of AA action allows reconciling the stimulatory action of AA on osteoclastogenesis observed in vitro with an overall attenuation of bone resorption in the presence of AA observed in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Ascorbic Acid / pharmacology*
  • Bone Marrow Cells / cytology*
  • Bone Marrow Cells / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Glutathione Disulfide / metabolism
  • Hydrogen Peroxide / metabolism
  • Hydrogen-Ion Concentration
  • Lactic Acid / metabolism
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mitochondria / metabolism
  • Osteoclasts / cytology*
  • Osteoclasts / drug effects*
  • Osteogenesis / drug effects


  • Lactic Acid
  • Adenosine Triphosphate
  • Hydrogen Peroxide
  • Ascorbic Acid
  • Glutathione Disulfide