Fecskeova Livia K, Matejova Jana, Slovinska Lucia, Bzdilova Jana, Kozovská Zuzana, Harvanova Denisa
Associated Tissue Bank, Faculty of Medicine, P. J. Safarik University and L. Pasteur University Hospital in Kosice, Tr. SNP 1, Kosice, 04011, Slovakia.
Cancer Research Institute, Biomedical Research Center SAS, Dúbravská cesta 9, Bratislava, 845 05, Slovakia.
Stem Cell Res Ther. 2025 Aug 5;16(1):423. doi: 10.1186/s13287-025-04542-9.
Extracellular vesicles (EVs) are the foundation of modern regenerative medicine using a cell-free approach. While current research mainly explores EVs from biological fluids and cell culture supernatants, tissue-derived EVs hold great promise, but remain largely underexplored. Since healthy placental tissues such as the chorion are widely available after full-term delivery, ethically unobjectionable, and possess exceptional regenerative potential, we sought to compare the biological effects of EVs derived directly from chorion tissue with those from chorion-derived mesenchymal stromal cell EVs and plasma EVs.
We compared the biological impact of EVs from various sources (chorion tissue CHO-Ti, MSCs from chorion CHO-MSC and platelet-poor plasma PPP) and isolated by various techniques on the gene expression of osteoarthritic chondrocytes. Additionally, we assessed the effect of enriched soluble proteins of CHO-MSC and CHO-Ti secretome vs. their EVs. EVs were characterized by particle number and size (NTA), protein content (BCA assay) and immunophenotype (flow cytometry). Changes in gene expression of chondrocytes were quantified by RT-qPCR.
CHO-Ti-EVs and PPP-EVs showed particularly beneficial effect on the inflammatory process, with their biological impact surpassing that of CHO-MSC-EVs. Chondroprotective markers COL2A and ACAN were robustly upregulated by CHO-Ti-EVs and PPP-EVs but showed only modest or variable increases with CHO-MSC-EVs. COMP expression, however, was specifically enhanced by CHO-MSC-derived components. Furthermore, our results also indicate that the therapeutic properties of the CHO-Ti secretome are exclusively linked to EVs. Among CHO-MSC-EVs, purification combined with UC resulted in the highest purity, however EVs purified by SEC presented a more favourable surface marker profile and better biological effects. The observed variability suggests that different EV preparations harbour distinct subpopulations that influence regulatory pathways differently and highlight the importance of EV source and isolation methodology in determining biological activity.
CHO-Ti-EVs showed promising effects on cartilage regeneration and inflammation modulation, suggesting they may represent a viable alternative to plasma- and CHO-MSC-EVs. Moreover, the chorion represents a readily accessible and abundant source of perinatal tissue obtainable non-invasively after full-term delivery, further supporting the translational potential of CHO-Ti-EVs.
细胞外囊泡（EVs）是采用无细胞方法的现代再生医学的基础。虽然目前的研究主要探索生物体液和细胞培养上清液中的EVs，但组织来源的EVs具有巨大潜力，但在很大程度上仍未得到充分研究。由于足月分娩后如绒毛膜等健康胎盘组织广泛可得，在伦理上无可非议，且具有卓越的再生潜力，我们试图比较直接源自绒毛膜组织的EVs与源自绒毛膜的间充质基质细胞EVs和血浆EVs的生物学效应。
我们比较了来自不同来源（绒毛膜组织CHO-Ti、来自绒毛膜的间充质干细胞CHO-MSC和少血小板血浆PPP）并通过各种技术分离的EVs对骨关节炎软骨细胞基因表达的生物学影响。此外，我们评估了CHO-MSC和CHO-Ti分泌组中富集的可溶性蛋白与其EVs的效果。通过颗粒数量和大小（NTA）、蛋白质含量（BCA测定）和免疫表型（流式细胞术）对EVs进行表征。通过RT-qPCR对软骨细胞基因表达的变化进行定量。
CHO-Ti-EVs和PPP-EVs对炎症过程显示出特别有益的作用，其生物学影响超过CHO-MSC-EVs。软骨保护标志物COL2A和ACAN被CHO-Ti-EVs和PPP-EVs强烈上调，但在CHO-MSC-EVs作用下仅显示适度或可变的增加。然而，COMP表达由CHO-MSC来源的成分特异性增强。此外，我们的结果还表明CHO-Ti分泌组的治疗特性仅与EVs相关。在CHO-MSC-EVs中，超滤结合超速离心产生的纯度最高，然而通过尺寸排阻色谱法纯化的EVs呈现出更有利的表面标志物谱和更好的生物学效应。观察到的变异性表明不同的EV制剂含有不同的亚群，它们对调节途径的影响不同，并突出了EV来源和分离方法在确定生物学活性方面的重要性。
CHO-Ti-EVs对软骨再生和炎症调节显示出有前景的效果，表明它们可能是血浆和CHO-MSC-EVs的可行替代物。此外，绒毛膜是足月分娩后可通过非侵入性获得的易于获取且丰富的围产期组织来源，进一步支持了CHO-Ti-EVs的转化潜力。
