Department of Orthodontics, Kunming Medical University School and Hospital of Stomatology, No.1088 Haiyuan Middle Road, Kunming, Yunnan 650106, China; Yunnan Key Laboratory of Stomatology, Kunming Medical University, 1168 Chunrong West Road, Kunming, Yunnan 650500, China; Center of Stomatology, Affiliated Hospital of Yunnan University, No.176 Qingnian Road, Kunming, Yunnan 650021, China.
Laboratory of Vaccine Development, Institute of Medical Biology, Chinese Academy of Medical Sciences, Peking Union Medical College, No.935 Jiaoling Road Kunming, Yunnan 650118, China.
Gene. 2025 Jan 15;933:148967. doi: 10.1016/j.gene.2024.148967. Epub 2024 Sep 26.
Static magnetic field (SMF) as an effective physical stimulus is capable of osteogenic differentiation for multiple mesenchymal stem cells, including human periodontal ligament stem cells (hPDLSCs). However, the exact molecular mechanism is still unknown. Therefore, this study intends to excavate molecular mechanisms related to SMF in hPDLSCs using functional experiments.
hPDLSCs were treated with different intensities of SMF, H19 lentivirus, and Wnt/β-catenin pathway inhibitor (XAV939). Changes in osteogenic markers (Runx2, Col Ⅰ, and BMP2), Wnt/β-catenin markers (β-catenin and GSK-3β), and calcified nodules were examined using RT-qPCR, western blotting, and alizarin red staining in hPDLSCs.
SMF upregulated the expression of H19, and SMF and overexpressing H19 facilitated the expression of osteogenic markers (Runx2, Col Ⅰ, and BMP2), activation of the Wnt/β-catenin pathway, and mineralized sediment in hPDLSCs. Knockdown of H19 alleviated SMF function, and treatment with XAV939 limited SMF- and H19-mediated osteogenic differentiation of hPDLSCs. Notably, the expression of hsa-miR-532-3p, hsa-miR-370-3p, hsa-miR-18a-5p, and hsa-miR-483-3p in hPDLSCs was regulated by SMF, and may form an endogenous competitive RNA mechanism with H19 and β-catenin.
SMF contributes to the osteogenic differentiation of hPDLSCs by mediating the H19/Wnt/β-catenin pathway, and hsa-miR-532-3p, hsa-miR-370-3p, hsa-miR-18a-5p, and hsa-miR-483-3p may be the key factors in it.
静磁场（SMF）作为一种有效的物理刺激，能够促进多种间充质干细胞的成骨分化，包括人牙周膜干细胞（hPDLSCs）。然而，其确切的分子机制尚不清楚。因此，本研究旨在通过功能实验挖掘 SMF 作用于 hPDLSCs 的相关分子机制。
用不同强度 SMF、H19 慢病毒和 Wnt/β-catenin 通路抑制剂（XAV939）处理 hPDLSCs。采用 RT-qPCR、western blot 和茜素红染色检测 hPDLSCs 中成骨标志物（Runx2、Col Ⅰ 和 BMP2）、Wnt/β-catenin 标志物（β-catenin 和 GSK-3β）和钙化结节的变化。
SMF 上调 H19 的表达，SMF 和过表达 H19 促进 hPDLSCs 中成骨标志物（Runx2、Col Ⅰ 和 BMP2）的表达、Wnt/β-catenin 通路的激活和矿化沉淀。H19 敲低减轻了 SMF 的作用，XAV939 处理限制了 SMF 和 H19 介导的 hPDLSCs 成骨分化。值得注意的是，hPDLSCs 中 hsa-miR-532-3p、hsa-miR-370-3p、hsa-miR-18a-5p 和 hsa-miR-483-3p 的表达受 SMF 调节，可能与 H19 和 β-catenin 形成内源性竞争 RNA 机制。
SMF 通过调节 H19/Wnt/β-catenin 通路促进 hPDLSCs 的成骨分化，hsa-miR-532-3p、hsa-miR-370-3p、hsa-miR-18a-5p 和 hsa-miR-483-3p 可能是其中的关键因素。
