Zhao Zhi-Meng, Ding Jia-Ming, Li Yu, Wang Da-Chuan, Kuang Ming-Jie
Department of Orthopedics, Shandong Provincial Hospital, Shandong University, Jinan, Shandong 250021, People's Republic of China.
Department of Orthopedics, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong 250021, People's Republic of China.
Stem Cells Transl Med. 2025 Mar 18;14(3). doi: 10.1093/stcltm/szae096.
Glucocorticoid-induced osteoporosis (GIOP), the most common cause of secondary osteoporosis, is characterized by significant bone loss, decreased bone quality, and increased fracture risk. The current treatments for GIOP have several drawbacks. Exosomes are vital for cellular processes. However, very few studies have focused on using human umbilical cord mesenchymal stem cell-derived exosomes (hUCMSC-EXOs) for GIOP treatment. In vitro and in vivo dexamethasone was used to evaluate the therapeutic effects of hUCMSC-EXOs on GIOP. CCK-8 and EdU assays were used to evaluate cell viability and proliferation, respectively. We conducted an alkaline phosphatase activity assay, alizarin red staining, Western blotting, and real-time PCR to detect the effect on osteogenesis. TMT-labeled quantitative proteomic and bioinformatic analyses were performed. Furthermore, we performed Western blotting, immunofluorescence, reactive oxygen species assays, and lipid peroxidation assays to investigate the regulatory mechanism by which hUCMSC-EXOs affect cell proliferation and osteogenic differentiation. The in vivo effects of hUCMSC-EXOs were evaluated using micro-CT, hematoxylin, and eosin staining, and immunohistochemical staining. We found that hUCMSC-EXOs reversed the inhibitory effects of glucocorticoids on human bone marrow stromal cell (hBMSC) proliferation and osteogenic differentiation and demonstrated that hUCMSC-EXOs reversed GIOP via the PI3K/AKT signaling pathway, inhibiting lipid peroxidation in vitro and in vivo. HUCMSC-EXOs promote hBMSC osteogenesis through the PI3K/AKT signaling pathway, inhibit ferroptosis, and have therapeutic potential for GIOP in mice.
糖皮质激素性骨质疏松症(GIOP)是继发性骨质疏松症最常见的病因,其特征为显著的骨质流失、骨质量下降和骨折风险增加。目前针对GIOP的治疗方法存在若干缺点。外泌体对细胞过程至关重要。然而,极少有研究聚焦于使用人脐带间充质干细胞衍生的外泌体(hUCMSC-EXOs)来治疗GIOP。在体外和体内使用地塞米松来评估hUCMSC-EXOs对GIOP的治疗效果。分别使用CCK-8和EdU检测法来评估细胞活力和增殖。我们进行了碱性磷酸酶活性检测、茜素红染色、蛋白质印迹法和实时聚合酶链反应,以检测对成骨作用的影响。进行了TMT标记的定量蛋白质组学和生物信息学分析。此外,我们进行了蛋白质印迹法、免疫荧光、活性氧检测和脂质过氧化检测,以研究hUCMSC-EXOs影响细胞增殖和成骨分化所涉及调节机制。使用显微计算机断层扫描、苏木精和伊红染色以及免疫组织化学染色来评估hUCMSC-EXOs的体内作用。我们发现hUCMSC-EXOs逆转了糖皮质激素对人骨髓间充质干细胞(hBMSC)增殖和成骨分化的抑制作用,并证明hUCMSC-EXOs通过PI3K/AKT信号通路逆转GIOP,在体外和体内抑制脂质过氧化。HUCMSC-EXOs通过PI3K/AKT信号通路促进hBMSC成骨,抑制铁死亡,对小鼠GIOP具有治疗潜力。