Abstract: This study aimed to identify and compare the miRNA profiles of exosomes derived from human iPSCs, BMSCs, and ADSCs (hiPSC-Exos, hBMSC-Exos, and hADSC-Exos) and their functional effects on human articular chondrocytes (hACs). hiPSC-Exos, hBMSC-Exos, and hADSC-Exos were collected from the appropriate cells cultured in 10% bovine exosome-depleted FBS (de-Exo-FBS) for 48 hours. NGS and bioinformatics were used to analyze the small RNA profiles of these exosomes. The biological functions of hACs were examined after a 12-day treatment with exosomes. hBMSC-Exos and hADSC-Exos had similar miRNA profiles but were largely different from hiPS-Exos. There were 17 highly expressed miRNAs in hiPSC-Exos, 13 miRNAs in hADSC-Exos, and 11 miRNAs in hBMSC-Exos. Among them, 7 miRNAs overlapped between the hBMSC-Exos and hADSC-Exos, and only 3 of them (hsa-miR-16-5p, hsa-miR-25-3p, and hsa-miR-93-5p) overlapped among all 3 exosomes. The putative target genes of the 3 overlapping exosomal miRNAs, and high-scoring target genes, including MAN2A1, ZNFX1, PHF19, GPR137C, ENPP5, B3GALT2, FNIP1, PKD2, and FBXW7, were identified. GO and KEGG enrichment analyses revealed that these genes are involved in cell growth, bone ossification, and cartilage development/differentiation, possibly via the MAPK signaling pathway. Accordingly, we confirmed the biological effect on cartilage differentiation and found that hiPSC-Exos, hBMSC-Exos and hADSC-Exos maintained hAC viability, prevented senescence, promoted the formation of a normal cartilage matrix (glycosaminoglycan and type II collagen), and downregulated fibrocartilage matrix (type I collagen) in normal hACs. Comparatively, hBMSC-Exos had the greatest effect on hAC function. Bioinformatics revealed differences and possible mechanisms of action of exosomes derived from pluripotent hiPSCs, multipotent hADSCs and multipotent hBMSCs and these exosomes effectively suppressed cell senescence and promoted normal functional extracellular matrix formation in hACs. Further investigations of the different functions of exosomes from pluripotent-hiPSCs other than those from multipotent-hMSCs are needed.
