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Osteoclast differentiation is an essential process that leads to bone loss, which is a hallmark of osteoporosis. In recent years, the use of hydrogen water (H2 water) has been proposed as a potential treatment for osteoporosis and other bone-loss related diseases. The aim of this study, published in the Journal of Bone and Mineral Metabolism, was to evaluate the effects of hydrogen molecules on RANKL-induced osteoclast differentiation in murine RAW264.7 cells.


The study used RAW264.7 cells, a mouse monocyte/macrophage cell line, and bone marrow-derived macrophages (BMMs) to investigate the effects of H2 on osteoclast differentiation. Cells were treated with H2, and the researchers measured the formation of resorption pits, the expression of osteoclast-specific markers, and intracellular reactive oxygen species (ROS) formation. They also investigated the activity of NADPH oxidase, Rac1, and Nox1, as well as mitochondrial ROS formation and the translocation of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) activity. In addition, the researchers looked at the activation of the mitogen-activated protein kinase (MAPK) and AKT pathways, as well as the activation of the nuclear factor-kappa B (NF-kB) pathway.


The study found that treatment with H2 prevented RANKL-induced osteoclast differentiation in RAW264.7 cells and BMMs. Treatment with H2 also inhibited the ability to form resorption pits in BMMs stimulated by RANKL. H2 treatment reduced mRNA levels of osteoclast-specific markers and decreased intracellular ROS formation, NADPH oxidase activity, Rac1 activity, and Nox1 expression. H2 treatment also increased nuclear translocation of Nrf2 and HO-1 activity, while suppressing the expression of nuclear factor of activated T cells c1 and c-Fos. In addition, H2 treatment suppressed the activation of the NF-kB pathway and reduced phosphorylation of p38, extracellular signal-regulated kinase (ERK), c-Jun-N-terminal kinase (JNK), and protein kinases B (AKT) stimulated by RANKL.


The study suggests that hydrogen molecules prevent RANKL-induced osteoclast differentiation, associated with inhibition of ROS formation and inactivation of the MAPK, AKT and NF-kB pathways. These findings provide new insights into the underlying molecular mechanisms of H2 water in preventing osteoclast differentiation and bone loss. However, further research is needed to confirm these findings in human subjects, and to explore the potential of H2 water as a therapeutic treatment for osteoporosis and other bone-loss related diseases.

Li, D.Z., et al., Treatment with hydrogen molecules prevents RANKL-induced osteoclast differentiation associated with inhibition of ROS formation and inactivation of MAPK, AKT and NF-kappa B pathways in murine RAW264.7 cells. J Bone Miner Metab, 2013.