Shinohara Issei, Susuki Yosuke, Murayama Masatoshi, Gao Qi, Cekuc Mehmet Sertac, Ergul Yasemin Sude, Morita Mayu, Pius Alexa K, Ma Chao, Chow Simon Kwoon-Ho, Goodman Stuart B
Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, CA, USA.
Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, Kobe, Hyogo, Japan.
HSS J. 2025 Jul 11:15563316251351031. doi: 10.1177/15563316251351031.
Long-term use of corticosteroids is a known risk factor for various bone diseases. Corticosteroids disrupt the balance between oxidative and glycolytic energy metabolism, increase oxidative stress and reactive oxygen species (ROS) associated with prolongation of inflammation, cell apoptosis, deficits in mesenchymal stem cells (MSCs), and osteoclast differentiation. Metformin, a drug for diabetes, has antioxidant properties by inhibiting nicotinamide adenine dinucleotide phosphate oxidase, which promotes the production of ROS.
We sought to evaluate the effects of corticosteroid and metformin administration on MSCs in vitro.
Primary bone marrow MSCs were collected from 20 mice. We evaluated prednisolone's effects on cell proliferation, oxidative stress, osteogenic differentiation, and mineralization, followed by metformin's effect on corticosteroid-induced reduction in bone formation. Metformin (1, 10, 100 µM) was tested with prednisolone 3 ng/mL. Cytokines were assessed by Luminex.
Prednisolone at 3 ng/mL significantly reduced cell proliferation, while 10 µM metformin restored it. Prednisolone increased oxidative stress and was reversed by metformin in a concentration-dependent manner, particularly at 100 µM. Osteogenic differentiation and mineralization were significantly impaired with prednisolone but improved with metformin at 10 and 100 µM. As for inflammatory cytokines, interleukin-1β (IL-1β) expression was increased by prednisolone administration and suppressed by metformin. Conversely, IL-6 and monocyte chemotactic protein-1 were suppressed by prednisolone.
This in vitro study found that corticosteroid-associated decrease in osteogenic potential of murine MSCs was associated with elevated oxidative stress that can be alleviated by metformin; further studies are needed to validate these findings in vivo and with human-derived MSCs.
长期使用皮质类固醇是导致各种骨疾病的已知风险因素。皮质类固醇会破坏氧化和糖酵解能量代谢之间的平衡，增加与炎症延长、细胞凋亡、间充质干细胞（MSC）缺陷和破骨细胞分化相关的氧化应激和活性氧（ROS）。二甲双胍是一种治疗糖尿病的药物，通过抑制促进ROS产生的烟酰胺腺嘌呤二核苷酸磷酸氧化酶而具有抗氧化特性。
我们试图评估皮质类固醇和二甲双胍给药对体外培养的间充质干细胞的影响。
从20只小鼠中收集原代骨髓间充质干细胞。我们评估了泼尼松龙对细胞增殖、氧化应激、成骨分化和矿化的影响，随后评估了二甲双胍对皮质类固醇诱导的骨形成减少的影响。用3 ng/mL泼尼松龙测试二甲双胍（1、10、100 μM）。通过Luminex检测细胞因子。
3 ng/mL的泼尼松龙显著降低细胞增殖，而10 μM二甲双胍可使其恢复。泼尼松龙增加氧化应激，二甲双胍以浓度依赖的方式逆转这种作用，尤其是在100 μM时。泼尼松龙显著损害成骨分化和矿化，但10和100 μM的二甲双胍可改善这种情况。至于炎性细胞因子，泼尼松龙给药会增加白细胞介素-1β（IL-1β）表达，而二甲双胍可抑制该表达。相反，泼尼松龙可抑制IL-6和单核细胞趋化蛋白-1。
这项体外研究发现，皮质类固醇导致的小鼠间充质干细胞成骨潜能降低与氧化应激升高有关，而二甲双胍可缓解这种氧化应激；需要进一步研究以在体内和人源间充质干细胞中验证这些发现。
