Mendiratta Meenakshi, Mendiratta Mohini, Sharma Yashvi, Sahoo Ranjit Kumar, Malhotra Neena, Mohanty Sujata
Stem Cell Facility, DBT-Centre of Excellence for Stem Cell Research, All India Institute of Medical Sciences, New Delhi, 110029, India.
Department of Medical Oncology, Dr. B. R. Ambedkar Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, New Delhi, 110029, India.
Stem Cell Res Ther. 2025 Jun 7;16(1):290. doi: 10.1186/s13287-025-04370-x.
Mesenchymal Stem Cells-derived Small Extracellular Vesicles endowed with regenerative cargo from their parent cells, have emerged as a promising avenue for cell-free therapeutics in regenerative medicine. Notably, deliberate induction of apoptosis in MSCs before sEV isolation has been identified as a strategy to augment the regenerative capabilities of MSCs-sEVs. This study explores a novel approach to enhance the immunomodulatory potential of MSC-sEVs through apoptosis induction and optimal tissue source to ensure consistent and improved clinical outcomes.
Apoptosis was induced in tissue-specific MSCs using Staurosporine. sEVs and sEVs were isolated via ultracentrifugation. Invitro immune response was assessed via T-cell proliferation, T-regulatory cell induction & macrophage polarization assay. Mitochondrial bioenergetics was studied using MitoSOX staining and Seahorse assay in H2O2-treated HuH7 cells. These findings were validated invivo in the CCL4-induced Chronic Liver Disease model via Histopathological staining, biochemical parameters, and fibrotic, pro-inflammatory, and anti-inflammatory markers and assessed the mechanism by targeting TGF-β/SMAD pathway.
Our results demonstrate that sEVs exhibited significantly higher concentrations and superior immunomodulatory effects by suppressing CD3 + T-cell proliferation, promoting T-regulatory cell differentiation, and polarized macrophages towards M2-phenotype. In terms of tissue specificity, it was observed that WJ-sEVs were faring better. sEVs effectively reduced mitochondrial ROS & significantly improved oxidative phosphorylation. Invitro findings were corroborated in an invivo CLD model, wherein sEVs ameliorated fibrosis and inflammation, by inhibiting TGF-β/ SMAD2/3 pathway.
This study concludes that apoptosis induction can be considered as minimum manipulation strategy to enhance the immunoregulatory and regenerative potential of MSCs-sEVs, thereby expanding their implication in immune disorder.
间充质干细胞衍生的小细胞外囊泡携带来自其母细胞的再生物质，已成为再生医学中无细胞治疗的一个有前景的途径。值得注意的是，在分离小细胞外囊泡之前有意诱导间充质干细胞凋亡已被确定为增强间充质干细胞小细胞外囊泡再生能力的一种策略。本研究探索了一种新方法，通过凋亡诱导和优化组织来源来增强间充质干细胞小细胞外囊泡的免疫调节潜力，以确保获得一致且改善的临床结果。
使用星形孢菌素在组织特异性间充质干细胞中诱导凋亡。通过超速离心分离小细胞外囊泡。通过T细胞增殖、T调节细胞诱导和巨噬细胞极化试验评估体外免疫反应。在H2O2处理的HuH7细胞中，使用MitoSOX染色和海马试验研究线粒体生物能量学。这些发现通过组织病理学染色、生化参数以及纤维化、促炎和抗炎标志物在CCL4诱导的慢性肝病模型中进行体内验证，并通过靶向TGF-β/SMAD途径评估机制。
我们的结果表明，小细胞外囊泡通过抑制CD3+T细胞增殖、促进T调节细胞分化以及使巨噬细胞向M2表型极化，表现出显著更高的浓度和更强的免疫调节作用。在组织特异性方面，观察到脐带华通氏胶间充质干细胞来源的小细胞外囊泡表现更好。小细胞外囊泡有效降低了线粒体活性氧并显著改善了氧化磷酸化。体外研究结果在体内慢性肝病模型中得到证实，其中小细胞外囊泡通过抑制TGF-β/SMAD2/3途径改善了纤维化和炎症。
本研究得出结论，凋亡诱导可被视为增强间充质干细胞小细胞外囊泡免疫调节和再生潜力的最小操作策略，从而扩大其在免疫疾病中的应用。
