Department of Joint Surgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
J Cell Biochem. 2019 Jan;120(1):171-181. doi: 10.1002/jcb.27289. Epub 2018 Sep 11.
The aim of the current study was to compare the expression of microRNAs (miRNAs) in exosomes derived from human bone mesenchymal stem cells (hBMSCs) with and without chondrogenic induction. Exosomes derived from hBMSCs were isolated and identified. Microarray analysis was performed to compare miRNA expression between exosomes derived from hBMSCs with and without chondrogenic induction, and quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the differentially expressed miRNAs. hBMSCs were transfected with miRNA mimic to extract miRNA-overexpressed exosomes. The results showed that most exosomes exhibited a cup-shaped or round-shaped morphology with a diameter of approximately 50-200 nm and expressed CD9 and CD63. We detected 141 miRNAs that were differentially expressed with and without chondrogenic induction by over a twofold change, including 35 upregulated miRNAs, such as miR-1246, miR-1290, miR-193a-5p, miR-320c, and miR-92a, and 106 downregulated miRNAs, such as miR-377-3p and miR-6891-5p. qRT-PCR analysis validated these results. Exosomes derived from hBMSCs overexpressing miR-320c were more efficient than normal exosomes derived from control hBMSCs at promoting osteoarthritis chondrocyte proliferation, down-regulated matrix metallopeptidase 13 and up-regulated (sex determining region Y)-box 9 expression during hBMSC chondrogenic differentiation. In conclusion, we identified a group of upregulated miRNAs in exosomes derived from hBMSCs with chondrogenic induction that may play an important role in mesenchymal stem cell-derived exosomes in cartilage regeneration and, ultimately, the treatment of arthritis. We demonstrated the potential of these modified exosomes in the development of novel therapeutic strategies.
本研究旨在比较诱导成软骨分化前后人骨髓间充质干细胞（hBMSCs）来源的外泌体中微小 RNA（miRNA）的表达。分离并鉴定 hBMSCs 来源的外泌体。进行微阵列分析比较诱导成软骨前后 hBMSCs 来源外泌体的 miRNA 表达，并使用定量实时聚合酶链反应（qRT-PCR）验证差异表达的 miRNA。用 miRNA 模拟物转染 hBMSCs 以提取 miRNA 过表达的外泌体。结果显示，大多数外泌体呈杯状或圆形，直径约为 50-200nm，并表达 CD9 和 CD63。我们检测到 141 个在诱导成软骨前后差异表达超过两倍的 miRNA，包括上调的 miRNA 如 miR-1246、miR-1290、miR-193a-5p、miR-320c 和 miR-92a，以及下调的 miRNA 如 miR-377-3p 和 miR-6891-5p。qRT-PCR 分析验证了这些结果。与正常对照 hBMSCs 来源的外泌体相比，过表达 miR-320c 的 hBMSCs 来源外泌体在促进骨关节炎软骨细胞增殖、下调基质金属蛋白酶 13 和上调（性别决定区 Y）-盒 9 表达方面更有效。总之，我们鉴定了一组诱导成软骨分化后的 hBMSCs 来源外泌体中上调的 miRNA，它们可能在间充质干细胞来源的外泌体在软骨再生和关节炎治疗中发挥重要作用。我们证明了这些修饰的外泌体在开发新的治疗策略方面的潜力。
