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脂肪间充质基质细胞经白细胞介素 1β处理后产生具有软骨保护作用的囊泡，能够快速穿透软骨。

Adipose-Derived Mesenchymal Stromal Cells Treated with Interleukin 1 Beta Produced Chondro-Protective Vesicles Able to Fast Penetrate in Cartilage.

机构信息

Laboratorio di Biotecnologie Applicate all'Ortopedia, IRCCS Istituto Ortopedico Galeazzi, I-20161 Milano, Italy.

INRIM-Istituto Nazionale di Ricerca Metrologica, 10135 Torino, Italy.

出版信息

Cells. 2021 May 12;10(5):1180. doi: 10.3390/cells10051180.

DOI:10.3390/cells10051180
PMID:34066077
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8151616/
Abstract

The study of the miRNA cargo embedded in extracellular vesicles (EVs) released from adipose-derived mesenchymal stromal cells (ASC) preconditioned with IL-1β, an inflammatory stimulus driving osteoarthritis (OA), along with EVs-cartilage dynamic interaction represent poorly explored fields and are the purpose of the present research. ASCs were isolated from subcutaneous adipose tissue and EVs collected by ultracentrifugation. Shuttled miRNAs were scored by high-throughput screening and analyzed through bioinformatics approach that predicted the potentially modulated OA-related pathways. Fluorescently labeled EVs incorporation into OA cartilage explants was followed in vitro by time-lapse coherent anti-Stokes Raman scattering; second harmonic generation and two-photon excited fluorescence. After IL-1β preconditioning, 7 miRNA were up-regulated, 4 down-regulated, 37 activated and 17 silenced. Bioinformatics allowed to identify miRNAs and target genes mainly involved in Wnt, Notch, TGFβ and Indian hedgehog (IHH) pathways, cartilage homeostasis, immune/inflammatory responses, cell senescence and autophagy. As well, ASC-EVs steadily diffuse in cartilage cells and matrix, reaching a plateau 16 h after administration. Overall, ASCs preconditioned with IL-1β allows secretion of EVs embedded with a chondro-protective miRNA cargo, able to fast penetrate in collagen-rich areas of cartilage with tissue saturation in a day. Further functional studies exploring the EVs dose-effects are needed to achieve clinical relevance.

摘要

本研究旨在探讨白细胞介素-1β（IL-1β）预处理的脂肪间充质基质细胞（ASC）释放的细胞外囊泡（EVs）中嵌入的微小 RNA（miRNA）货物，以及 EVs-软骨动态相互作用。这些领域尚未得到充分探索，是本研究的目的。从皮下脂肪组织中分离 ASC，并通过超速离心收集 EVs。通过高通量筛选对穿梭 miRNA 进行评分，并通过生物信息学方法分析预测潜在调节骨关节炎（OA）相关途径。通过共聚焦反斯托克斯拉曼散射；二次谐波产生和双光子激发荧光，在体外观察荧光标记的 EVs 掺入 OA 软骨外植体。经 IL-1β预处理后，有 7 个 miRNA 上调，4 个下调，37 个激活，17 个沉默。生物信息学分析鉴定出 miRNA 和靶基因主要涉及 Wnt、Notch、TGFβ 和 Indian hedgehog（IHH）途径、软骨稳态、免疫/炎症反应、细胞衰老和自噬。此外，ASC-EVs 可稳定扩散到软骨细胞和基质中，给药 16 小时后达到平台期。总之，IL-1β 预处理的 ASC 可分泌富含软骨保护 miRNA 货物的 EVs，能够在一天内快速穿透富含胶原蛋白的软骨区域，并达到组织饱和。需要进一步进行探索 EVs 剂量效应的功能研究，以实现临床相关性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5950/8151616/4ea861fe48df/cells-10-01180-g005.jpg
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