Le Guoping, Wen Riyou, Fang Huaixi, Huang Zhifa, Wang Yong, Luo Hanwen
Department of Joint Osteopathy, Liuzhou Worker's Hospital, Liuzhou, Guangxi Province, 545000, China.
J Orthop Surg Res. 2025 Jan 29;20(1):107. doi: 10.1186/s13018-025-05515-7.
Alcoholic osteonecrosis of the femoral head (AIONFH) is caused by long-term heavy drinking, which leads to abnormal alcohol and lipid metabolism, resulting in femoral head tissue damage, and then pathological necrosis of femoral head tissue. If not treated in time in clinical practice, it will seriously affect the quality of life of patients and even require hip replacement to treat alcoholic femoral head necrosis. This study will confirm whether M2 macrophage exosome (M2-Exo) miR-122 mediates alcohol-induced BMSCs osteogenic differentiation, ultimately leading to the inhibition of femoral head necrosis. M2 macrophages were identified by flow cytometry, and the isolated exosomes were characterized by transmission electron microscopy (TEM) and Nanoparticle Tracking Analysis (NTA). Next, miR-122 was overexpressed by transfecting miR-122 mimic, and the expression of miR-122 in M2 macrophages and their exosomes was evaluated. Subsequently, the effect of exosomal miR-122 on the osteogenic differentiation ability of BMSCs was detected, including cell proliferation, expression of osteogenic-related genes (RUNX2, BMP2, OPN, ALP), and calcium nodule formation. Finally, the therapeutic effect of M2-Exo was analyzed in a rat model of AIONFH, and bone repair and pathological damage were evaluated by Micro-CT, RT-qPCR, HE, Masson staining, and immunohistochemistry (COL I). The results showed that M2 macrophages were successfully polarized, with an average M2-Exo particle size of 156.4 nm and a concentration of 3.2E + 12 particles/mL. The expression of miR-122 in M2 macrophages is significantly higher than that in M0 macrophages, and miR-122 mimic can increase the content of miR-122 in M2-Exo. miR-122 in M2-Exo can promote osteogenic differentiation of rat bone marrow BMSCs, enhance cell viability, and increase the expression of osteogenesis-related genes. After being applied to the AIONFH rat model, the injection of M2-exo and miR-122 mimics significantly improved the repair effect of articular cartilage, alleviated pathological changes, and promoted the regeneration of bone tissue. M2-macrophage-derived exosomal miR-122 induces osteogenic differentiation of bone mesenchymal stem cells in treating AIONFH.
酒精性股骨头坏死（AIONFH）是由长期大量饮酒引起的，这会导致酒精和脂质代谢异常，造成股骨头组织损伤，进而引发股骨头组织的病理性坏死。在临床实践中若不及时治疗，将严重影响患者的生活质量，甚至需要进行髋关节置换来治疗酒精性股骨头坏死。本研究将证实M2巨噬细胞外泌体（M2-Exo）中的miR-122是否介导酒精诱导的骨髓间充质干细胞（BMSCs）成骨分化，最终导致股骨头坏死受到抑制。通过流式细胞术鉴定M2巨噬细胞，采用透射电子显微镜（TEM）和纳米颗粒跟踪分析（NTA）对分离出的外泌体进行表征。接下来，通过转染miR-122模拟物使miR-122过表达，并评估miR-122在M2巨噬细胞及其外泌体中的表达。随后，检测外泌体miR-122对BMSCs成骨分化能力的影响，包括细胞增殖、成骨相关基因（RUNX2、BMP2、OPN、ALP）的表达以及钙结节形成。最后，在AIONFH大鼠模型中分析M2-Exo的治疗效果，并通过Micro-CT、RT-qPCR、苏木精-伊红（HE）染色、Masson染色和免疫组织化学（COL I）评估骨修复和病理损伤情况。结果显示，M2巨噬细胞成功极化，M2-Exo平均粒径为156.4 nm，浓度为3.2E+12颗粒/毫升。M2巨噬细胞中miR-122的表达显著高于M0巨噬细胞，且miR-122模拟物可增加M2-Exo中miR-122的含量。M2-Exo中的miR-122可促进大鼠骨髓BMSCs的成骨分化，增强细胞活力，并增加成骨相关基因的表达。将其应用于AIONFH大鼠模型后，注射M2-外泌体和miR-122模拟物可显著改善关节软骨的修复效果，减轻病理变化，并促进骨组织再生。M2巨噬细胞来源的外泌体miR-122在治疗AIONFH中可诱导骨间充质干细胞成骨分化。
