Fig. 1

thMSC-EVs significantly improved cell viability, cell integrity, and inflammation in NEC in vitro models.(A) Bar graph representation of CCK-8 cell viability assay, TUNEL assay, and western blot protein expression levels of ZO-1 in IEC-6 cells. n = 16, 16, 16, and 16 for the CCK-8 assay; n = 10, 10, 10, and 10 for the TUNEL assay; and n = 3, 3, 3, and 3 for western blot analysis in the CON, HIT, HIT + naïveEV, and HIT + thEV groups, respectively. *, p < 0.01 vs. CON; #, p < 0.01 vs. HIT; $, p < 0.01 vs. HIT + naïveEV. (B) Bar graph representation of CCK-8 cell viability assay in LPS-induced peritoneal macrophage CM-treated IEC-6 cells. n = 16, 16, 16, and 16 in the CON, LPS CM, LPS + naïveEV CM, and LPS + thEV CM groups, respectively. (C) Bar graph showing inflammatory cytokine levels in LPS-induced peritoneal macrophages. n = 4, 4, 4, and 4 in the CON, LPS, LPS + naïveEV, and LPS + thEV groups, respectively. *, p < 0.01 vs. CON; #, p < 0.01 vs. LPS; $, p < 0.01 vs. LPS + naïveEV. Data are presented as mean ± SEM. One-way ANOVA and the post-hoc Tukey test were used in the analysis. CON, normal control; HIT, hyperosmolar stress, ischemia, and hypothermia-induced IEC-6 cells; LPS, lipopolysaccharide; naïve MSC-EVs, naïve MSC-derived EVs; thMSC-EVs, thrombin-preconditioned MSC-derived EVs