Large extracellular vesicles secreted by human iPSC-derived MSCs ameliorate tendinopathy via regulating macrophage heterogeneity

• Published in: Bioactive materials 2023-03, Vol.21, p.194-208
• Main Authors: Ye, Teng, Chen, Zhengsheng, Zhang, Jieyuan, Luo, Lei, Gao, Renzhi, Gong, Liangzhi, Du, Yuhang, Xie, Zongping, Zhao, Bizeng, Li, Qing, Wang, Yang
• Format: Article
• Language: English
• Subjects: Inflammatory regulation; iPSC-derived MSCs; Large extracellular vesicles; Macrophages; Tendinopathy
• ISSN: 2452-199X; 2452-199X
• DOI: 10.1016/j.bioactmat.2022.08.007

Tendinopathy is a common musculoskeletal disorder which results in chronic pain and reduced performance. The therapeutic effect of stem cell derived-small extracellular vesicles (sEVs) for tendinopathy has been validated in recent years. However, whether large extracellular vesicles (lEVs), another subset of extracellular vesicles, possesses the ability for the improvement of tendinopathy remains unknown. Here, we showed that lEVs secreted from iPSC-derived MSCs (iMSC-lEVs) significantly mitigated pain derived from tendinopathy in rats. Immunohistochemical analysis showed that iMSC-lEVs regulated the heterogeneity of infiltrated macrophages and several inflammatory cytokines in rat tendon tissue. Meanwhile, in vitro experiments revealed that the M1 pro-inflammatory macrophages were repolarized towards M2 anti-inflammatory macrophages by iMSC-lEVs, and this effect was mediated by regulating p38 MAPK pathway. Moreover, liquid chromatography-tandem mass spectrometry analysis identified 2208 proteins encapsulated in iMSC-lEVs, including 134 new-found proteins beyond current Vesiclepedia database. By bioinformatics and Western blot analyses, we showed that DUSP2 and DUSP3, the negative regulator of p38 phosphorylation, were enriched in iMSC-lEVs and could be transported to macrophages. Further, the immunomodulatory effect of iMSC-lEVs on macrophages was validated in explant tendon tissue from tendinopathy patients. Taken together, our results demonstrate that iMSC-lEVs could reduce inflammation in tendinopathy by regulating macrophage heterogeneity, which is mediated via the p38 MAPK pathway by delivery of DUSP2 and DUSP3, and might be a promising candidate for tendinopathy therapy. [Display omitted] •iMSC-lEVs significantly ameliorate tendinopathy both in a rat model and explant tendon tissue from human patient.•iMSC-lEVs modulate macrophages polarization via p38 MAPK signaling pathway.•Proteomics analysis of iMSC-lEVs discovers a new set of 134 proteins beyond current Vesiclepedia Database.•The p38 MAPK signaling pathway-mediated macrophage repolarization was partly regulated by the delivery of DUSP2 and DUSP3.•The immunoregulatory function of iMSC-lEVs are similar with iMSC-sEVs.