NAD+/Nrf2 signaling promotes osteogenesis by regulating oxidative level of BMSCs under mechanical stress

作者：Huiying Ren, Jixiao Wang, Jiani Liu, Zijie Zhang, Lingyun Wang, Fulan Wei · 发表于：Progress in Orthodontics · 年份：2025 · DOI：10.1186/s40510-025-00566-2 · 被引用次数：5 · 研究领域：Mesenchymal stem cell research、RNA Interference and Gene Delivery、Antimicrobial Peptides and Activities

Abstract Background Mechanical stress triggers an increase in cellular reactive oxygen species (ROS), which is associated with the impairment of osteogenesis. During orthodontic treatment, bone marrow mesenchymal stem cells (BMSCs) experience mechanical stress, yet the oxidative profile and redox regulatory mechanisms under such stress, especially involving Nicotinamide adenine dinucleotide (NAD + ), are not well understood, necessitating further research into their roles in orthodontic therapies. Methods The Tension System was established to detect ROS changes in BMSCs under cyclic stretch stress, with H 2 O 2 simulating uncontrolled ROS. Flow cytometry and fluorescence staining measured ROS, while an NAD + /NADH assay kit assessed NAD + levels. qRT-PCR and Western blotting analyzed expression of NAD + synthesis and consume enzymes. Osteogenic potential was evaluated by qRT-PCR, Western blotting, and Alkaline phosphatase (ALP) staining. Loss-of-function and supplementation assays explored role of NAD + in oxidative stress and Nrf2 regulation, with localization assessed by immunofluorescence and Western blotting. In vivo osteogenic effects were confirmed using an orthodontic tooth movement (OTM) model, with osteogenesis assessed by immunohistochemistry and microCT for OTM measurements. Results Cyclic stretch stress increased ROS in BMSCs over 24 h and boosted osteogenic differentiation. However, increased ROS from H 2 O 2 hindered this process. Notably, NAD + levels rose with...