Hydrogen (H₂) is one of the major biodegradation products of magnesium (Mg) alloys implanted for bony fracture healing and reconstruction; H₂ thus plays a significant role in the regulation of local microenvironment and the biology of resident cells. The interactions between the H₂ and the local cells are of great interest, and a full understanding of the effect of H₂ on bone marrow mononuclear cells (BMMCs) would accelerate the development of effective strategies for successful bony healing. This study investigates how H₂, with different concentrations and durations, regulates the osteoclastogenesis of mouse BMMCs. First, using H₂ with five concentrations (0%, 2%, 25%, 50% and 75%) and three durations (5, 7 and 10 days), the osteoclastogenesis of mouse BMMCs in these H₂ conditions were measured using TRAP staining, F-actin ring formation assay, pit formation assay and RT-qPCR analysis. Based on these findings, the proliferation assay, apoptosis assay, western blot analysis and ELISA assay of BMMCs after osteoclast induction were performed. The findings showed that H₂ (especially the 50% and 75% H₂) obviously inhibited the osteoclast formation, function and osteoclast-related genes expression of osteoclast-induced BMMCs; additionally, H₂ (50%) was found to reduce the proliferation, promote the apoptosis and inhibit the expression of osteoclast-related proteins of BMMCs with the presence of osteoclast-induced medium. Therefore, H₂ significantly inhibited the osteoclastogenesis of mouse BMMCs, which may become a new therapeutic agent for anti-bony resorption and open new avenues for the translational research of Mg alloys.

氢（H ₂）是植入用于骨性骨折愈合和重建的镁（Mg）合金的主要生物降解产物之一。因此，H ₂在调节局部微环境和驻留细胞的生物学中起重要作用。H ₂与局部细胞之间的相互作用引起了极大的兴趣，并且对H ₂对骨髓单核细胞（BMMC）的影响的充分理解将加速成功骨愈合的有效策略的发展。这项研究调查了不同浓度和持续时间的H ₂如何调节小鼠BMMC的破骨细胞生成。首先，使用H ₂在五个浓度（0％，2％，25％，50％和75％）和三个持续时间（5、7和10天）的情况下，使用TRAP染色，F-肌动蛋白测量了在这些H ₂条件下小鼠BMMC的破骨细胞生成环形成分析，凹坑形成分析和RT-qPCR分析。基于这些发现，进行了破骨细胞诱导后BMMC的增殖测定，凋亡测定，蛋白质印迹分析和ELISA测定。研究结果显示使得h ₂（特别是50％和75％H ₂）明显抑制破骨细胞诱导的BMMCs的破骨细胞形成，功能和破骨细胞相关基因的表达; 另外，H ₂（50％）在破骨细胞诱导培养基的存在下，可降低BMMC的增殖，促进细胞凋亡并抑制破骨细胞相关蛋白的表达。因此，H ₂显着抑制小鼠BMMC的破骨细胞生成，可能成为抗骨吸收的新治疗剂，为镁合金的转化研究开辟新的途径。