{"paper_id":"3577246b-4f91-4e1d-8b04-d0a57d9a28c9","body_text":"Abstract\nEndometriosis is a benign yet chronic gynecological disorder characterized by dysregulation of processes such as inflammation, angiogenesis, migration, apoptosis, and proliferation. Menstrual blood-derived endometrial stem cells play a crucial role in the retrograde development and progression of endometriotic lesions. To evaluate the therapeutic potential of exosomes derived from menstrual blood-derived stem cells, exosomes from non-endometriotic MenSCs (NE-MenSCs), both unmodified (Exo) and transfected with miR-4289, were applied as treatments to MenSCs from the endometriosis cell line (E-MenSCs). Publicly available databases were used to identify key genes and signaling pathways implicated in endometriosis, from which miR-4289 was selected as an effective regulatory microRNA. Following treatment, cellular migration was assessed by scratch assays; gene expression was evaluated via real-time PCR; protein levels of ROS, IL-10, and IL-1β were measured by ELISA; and ESR1, CTNNB1, and Ki67 levels were determined by Western blotting. The results indicate that treatments significantly reduced the expression of genes associated with inflammation, proliferation, migration, and the Wnt/β-catenin pathway. Scratch assays and reductions in MMP9 expression suggest decreased migration in the Exo and miR-Exo groups. The expression of CTNNB1, IL-1β, and IL-10 was significantly downregulated in treated groups compared to E-MenSCs. In addition, KRAS and IDO1 expression levels were significantly decreased following treatment, and Ki67 protein levels were notably reduced in the miR and miR-Exo groups. These findings highlight the therapeutic potential of MenSC-derived exosomes loaded with miR-4289 as a promising and novel strategy for treating endometriosis.\nGraphical Abstract\nSimilar content being viewed by others\nData Availability\nThe datasets used and/or analyzed during the current study are available in supplementary section.\nAbbreviations\n- MenSCs:\n-\nMenstrual Stem Cells\n- NE-MenSCs:\n-\nNon-Endometriosis Menstrual Stem Cells\n- E-MenSCs:\n-\nEndometriosis Menstrual Stem Cells\n- Exo group:\n-\nExosome Group\n- miR-Exo group:\n-\nmicroRNA-Exosome Group\n- miR group:\n-\nmicroRNA Group\n- BB:\n-\nBackbone\n- EV:\n-\nExtracellular vesicle\n- DLS:\n-\nDynamic light scattering\n- CM:\n-\nConditioned medium\n- TEM:\n-\nTransmission electron microscopy\nReferences\nAgarwal A, Gupta S, Sharma RK (2005) Role of oxidative stress in female reproduction. 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All authors reviewed and approved the manuscript **.**.\nCorresponding author\nEthics declarations\nCompeting Interest\nThe authors declare no competing interests.\nEthical Approval and Consent to Participate\nThis experimental investigation conducted in the field of endometriosis, received ethical approval from the Islamic Azad University Ethics Committee (ethical code: IR.IAU.SRB.REC.1401.312). NE-MenSCs and E-MenSCs cell lines were procured from the Department of Cell Biology and Regenerative Medicine of ACECR, Qom Branch. Characterization of these cell lines was confirmed by flow cytometry, as reported in a published article (Asl et al. 2023). We then investigated treatments using exosomes and microRNA.\nConsent for Publication\nNot applicable.\nAdditional information\nPublisher’s Note\nSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\nSupplementary Information\nBelow is the link to the electronic supplementary material.\nRights and permissions\nSpringer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.\nAbout this article\nCite this article\nMahmoudi, S., Sheikholeslami, A., Roodbari, N.H. et al. Evaluating the Effect of Exosome-Encapsulated miR-4289 on Menstrual Blood-Derived Mesenchymal Stem Cells from Endometriosis Patients. Biochem Genet (2025). https://doi.org/10.1007/s10528-025-11265-2\nReceived:\nAccepted:\nPublished:\nVersion of record:\nDOI: https://doi.org/10.1007/s10528-025-11265-2","source_license":"CC0","license_restricted":false}