Pan Chun, Cheng Cheng, Zhong Shu, Li Shiyu, Tan Wei, Yao Yachao
The Affiliated Guangdong Second Provincial General Hospital of Jinan University, Guangzhou, 510317, China.
Institute of Genomic Medicine, College of Pharmacy, Jinan University, Guangzhou, 511436, China.
J Orthop Surg Res. 2025 Apr 17;20(1):388. doi: 10.1186/s13018-025-05749-5.
The secretion of mitochondrial-derived vesicles (MDVs) has been found to increase during osteogenic differentiation, but their role in intercellular communication and osteogenic promotion remains unclear. In this study, we extracted translocase of outer mitochondrial membrane 20 (Tomm20) + MDVs from bone marrow stromal cells (BMSCs) at different osteogenic culture days using differential centrifugation and immunoprecipitation, then co-cultured them with BMSCs to assess osteogenic differentiation, immune response and metabolic levels. The results showed that osteogenic differentiation enhances MDVs' secretion and their mitochondrial DNA (mtDNA) content. In promoting osteogenic differentiation ability, osteogenic-induced MDVs (MDV-OMs, especially MDV-OM14 and MDV-OM21) significantly enhance mineralization with OD values 1.37-fold and 1.32-fold higher than those of MDV-OM7 (p < 0.05) after 21 days, respectively. However, these MDVs containing mtDNA activate immune responses by upregulating cGas, Sting, Caspase-9, Il-6, and Tnf-a mRNA levels, inducing cell apoptosis and oxidative stress. In addition, MDVs containing mitochondrial components also have metabolic regulatory functions. Metabolic level detection revealed that MDV-OMs downregulate lactate, promote tricarboxylic acid cycle (TCA) enzyme expression, and increase mitochondrial membrane potential. Among these MDVs, MDV-OM7, induced for 7 days, shows osteogenic function without strong immune response, possibly related to metabolic reprogramming. This study highlights the potential of osteogenic-induced MDVs for bone regeneration, cGAS-STING activation, and metabolic enhancement, and are expected to be used for the treatment of diseases such as tissue damage.
已发现线粒体衍生囊泡（MDVs）的分泌在成骨分化过程中会增加，但其在细胞间通讯和成骨促进中的作用仍不清楚。在本研究中，我们使用差速离心和免疫沉淀法从不同成骨培养天数的骨髓间充质干细胞（BMSCs）中提取外线粒体膜转位酶20（Tomm20）+ MDVs，然后将它们与BMSCs共培养，以评估成骨分化、免疫反应和代谢水平。结果表明，成骨分化增强了MDVs的分泌及其线粒体DNA（mtDNA）含量。在促进成骨分化能力方面，成骨诱导的MDVs（MDV-OMs，尤其是MDV-OM14和MDV-OM21）在21天后显著增强矿化，其OD值分别比MDV-OM7高1.37倍和1.32倍（p < 0.05）。然而，这些含有mtDNA的MDVs通过上调cGas、Sting、Caspase-9、Il-6和Tnf-a mRNA水平激活免疫反应，诱导细胞凋亡和氧化应激。此外，含有线粒体成分的MDVs也具有代谢调节功能。代谢水平检测显示，MDV-OMs下调乳酸水平，促进三羧酸循环（TCA）酶表达，并增加线粒体膜电位。在这些MDVs中，诱导7天的MDV-OM7显示出成骨功能，而没有强烈的免疫反应，这可能与代谢重编程有关。本研究突出了成骨诱导的MDVs在骨再生、cGAS-STING激活和代谢增强方面的潜力，并有望用于治疗组织损伤等疾病。
