Exosomes derived from 3D-cultured hUCMSCs exhibit superior hepatoprotection against ALF

Most recently, exosomes (Exos) derived from 3D-cultured cells exhibit great advantages over 2D-Exos. Nevertheless, it remains to be explored whether 3D (tissue engineering liver) exosomes are superior to 2D-Exos in protecting mice from acute liver injury (ALF) and the underlying mechanism. Herein, we demonstrated that 3D-Exos manifested higher yields, protein concentration and RNA content compared to 2D-Exos. Remarkably, 3D-Exos exhibited superior hepatoprotection against ALF, as shown by reduced TBiL levels, improved liver architecture and alleviated inflammation. According to miRNA sequencing and KEGG enrichment analysis, the differentially expressed genes between 2D- and 3D-Exos could be enriched in "Hippo-YAP" and "autophagy" pathways. Moreover, the levels of YAP/TAZ and autophagy markers were more elevated in 3D-Exos-treated mice. Specifically, inhibiting YAP and autophagy signaling destroyed the hepatoprotection conferred by 3D-Exos. Collectively, 3D-Exos exert superior hepatoprotection against ALF by up-regulating YAP and autophagy signaling. Our finding provides powerful support for administering 3D-Exos to treat ALF efficiently.