biostudies-literatureUnknown28(9)Tang HMost 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 markiScience113419https://www.ebi.ac.uk/biostudies/studies/S-EPMC12446390biostudies-literatureExosomes derived from 3D-cultured hUCMSCs exhibit superior hepatoprotection against ALF.PMC12446390Liu XDuan ZXu MLu WLi SZhang YBai LChen YYang QTang HfalseExosomes 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 mark2025-01-01T00:00:00Z2025 Sep2026-06-03T17:28:28.354Z2026-04-29T03:13:26.487ZS-EPMC124463904097815110.1016/j.isci.2025.113419