Expression of exosomal microRNAs during chondrogenic differentiation of human bone mesenchymal stem cells

Abstract

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.

Extraction and identification of synovial tissue-derived exosomes by different separation techniques

“…This suggests that synovial tissue may regulate chondrocytes by secreting exosomes. Number of articles [15][16][17][18][19][20][21][22] has conducted detailed studies on how to extract cell supernatants and exosomes in body fluids, but few of articles had studied the extraction and identification of exosomes in tissues. To the best of our knowledge, any article has been published on how to extract and identify exosomes from synovial tissue.…”

Articular chondrocyte-derived exosomes promote cartilage differentiation of human umbilical cord mesenchymal stem cells by activation of autophagy

“…Exosomes, as important paracrine components, are involved in maintaining normal physiological functions, mediating inter-cell communication, and inducing changes in cell functions and processes by delivering various types of bioactive microRNAs, proteins, and unique gene products 9,10,24 . These microRNAs might serve as vital inducers of HUCMSC differentiation into chondrocytes, such as the exosomal miR-92a-3p, exosomal miR-95-5p, exosomal miR-320c, and exosomal miR-135b, which can regulate cartilage development and homeostasis [25][26][27][28] . These ndings suggest that the active molecules in exosomes may be an important mechanism underlying their ability to promote chondrogenesis of HUCMSCs.…”

Exosomes in osteoarthritis and cartilage injury: advanced development and potential therapeutic strategies

“…There are two ways to load drugs into exosomes, one is to load drugs into the donor cell of exosomes, such as using transfection and co-incubation; the other is to load drugs into exosomes after they are secreted, such as direct mixing, which the loading efficacy is a big concern. At present, most researchers prefer to obtain the exosomes for OA therapy with high expression of miRNA or lncRNA from modified MSCs [22,[44][45][46] . Exosomal miR-92a-3p [22] , exosomal lncRNA-KLF3-AS1 [44] , exosomal miR-140-5p [45] and exosomal miR-320c [46] from transfected MSCs have been reported to have significant therapeutic effects on OA in vivo and in vitro.…”

“…At present, most researchers prefer to obtain the exosomes for OA therapy with high expression of miRNA or lncRNA from modified MSCs [22,[44][45][46] . Exosomal miR-92a-3p [22] , exosomal lncRNA-KLF3-AS1 [44] , exosomal miR-140-5p [45] and exosomal miR-320c [46] from transfected MSCs have been reported to have significant therapeutic effects on OA in vivo and in vitro. Furthermore, it was reported that primary chondrocytes also could be modified to serve as the donor cells of exosomes [24] .…”

“…The cell-friendly biological feature of exosome decides that the future drug application via exosome may provide a higher delivery efficiency. The recent research tend is to modify the exosome-secreting cells with transfection to load miRNA(s) into exosomes, thus can be used for OA therapy/cartilage repair [23,[44][45][46] . However, the transfection method in loading RNA has many limitations, such as the unstable productivity of RNA and the unclear factors influencing the RNA level and loading [75] , which means the application is still far from clinic.…”