Lotfi Maryam, Baharara Javad, Shahrokhabadi Khadije Nejad, Khorshid Pejman
Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran.
Department of Biology & Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran.
Curr Stem Cell Res Ther. 2025;20(3):336-349. doi: 10.2174/011574888X314834240628110545.
Cartilage has intrinsically limited healing power, and regeneration of cartilage damages has remained a challenge. Secreted products of mesenchymal stem cells have shown a new therapeutic strategies for cartilage injuries. Also it has been observed that low frequency electromagnetic field plays a key role in biological processes.
This research was performed to investigate the synergic effect of mesenchymal stem cell-derived exosomes and low frequency electromagnetic field on chondrogenic differentiation.
In this study, mesenchymal stem cell-derived exosomes were identified using AFM, SEM, TEM microscopy, and DLS method. Cells were treated in chondrogenic medium by exosomes, low frequency electromagnetic field, and the synergy of both. The cell survival was examined using MTT and Annexin methods, and cartilage differentiation was confirmed by Alcian blue staining. The expression of Sox-9, Acan, Col 2a1 and Col 10a1 genes was examined via Real- Time PCR technique on day 14 post-treatment.
The results confirmed the presence of exosomes with an approximate size of less than 100 nm. The results of Alcian blue revealed greater expression of glycosaminoglycans in the synergic treatment group compared to the other groups. Real-time PCR showed a significant increase in the expression of Sox-9, Acan, and Col 2a1 genes, as well as a significant reduction of Col 10a1 gene expression in the synergic treatment group compared to other groups.
This study indicated that the synergic effect of exosome and low-frequency electromagnetic fields would lead to enhanced chondrogenic differentiation, which can be further explored in future clinical studies.
软骨自身的愈合能力有限，软骨损伤的再生一直是一项挑战。间充质干细胞的分泌产物已显示出治疗软骨损伤的新策略。此外，人们还观察到低频电磁场在生物过程中起关键作用。
本研究旨在探讨间充质干细胞来源的外泌体与低频电磁场对软骨形成分化的协同作用。
在本研究中，使用原子力显微镜、扫描电子显微镜、透射电子显微镜和动态光散射法鉴定间充质干细胞来源的外泌体。细胞在软骨形成培养基中分别用外泌体、低频电磁场以及两者的协同作用进行处理。使用MTT法和膜联蛋白法检测细胞存活率，并用阿尔辛蓝染色法确认软骨分化。在处理后第14天，通过实时荧光定量PCR技术检测Sox-9、Acan、Col 2a1和Col 10a1基因的表达。
结果证实存在大小约小于100 nm的外泌体。阿尔辛蓝染色结果显示，与其他组相比，协同治疗组中糖胺聚糖的表达更高。实时荧光定量PCR显示，与其他组相比，协同治疗组中Sox-9、Acan和Col 2a1基因的表达显著增加，而Col 10a1基因的表达显著降低。
本研究表明，外泌体与低频电磁场的协同作用可增强软骨形成分化，这在未来的临床研究中值得进一步探索。
