Fig 2: Nintedanib (Nin) administration restored the expression of the neutrophil marker Ly6G, VLA-4 and VCAM-1 in the lungs of mice with LPS-induced ALI. (A–C) Intratracheal injection of LPS significantly increased the expression of Ly6G, VLA-4, and VCAM-1 in the lungs of mice with ALI compared with control mice. Nintedanib administration reversed these changes. Data are means ± standard deviations. * p < 0.05 vs. control, # p < 0.05 vs. LPS; n = 6 per group. PBS, phosphate-buffered saline; IHC, immunohistochemical stains.

Fig 3: Administration of LPS increased the expression of adhesion molecules (including VCAM-1, VLA-4, ICAM-1, and LFA-1) in lung tissue. Administration of miPSCs or miPSC-CM reduced adhesion molecules in mice with LPS-induced ALI. (a,b) Immunohistochemistry (IHC) staining of lung sections show that VCAM-1 expression significantly increased following the administration of LPS compared to control mice. In contrast, the expression of VCAM-1 in ALI mice was reduced by intravenous delivery of miPSCs or miPSC-CM. Similarly, the expression of VLA-4, ICAM-1, and LFA-1 in the lung was increased by LPS stimulation and was inhibited by administering miPSCs or miPSC-CM. Data are presented as the mean ± SD. * p < 0.05, compared to control mice; ** p < 0.05, compared to LPS-induced ALI mice; n = 6 per group.

Fig 4: Administration of hiPSCs or hiPSC-CM reduced human neutrophils (D-HL-60 cells) adhering to human endothelial cells (HUVECs) and inhibited the TEM of neutrophils induced by LPS. Administration of hiPSCs or hiPSC-CM reduced the VCAM-1 expression induced by LPS in HUVEC and VLA-4 induced by LPS in D-HL-60 cells. (a) The D-HL-60 cell/HUVEC adhesion assay showed that D-HL-60 cells adhering to HUVECs significantly increased after administering LPS compared to the control. In contrast, the D-HL-60 cell/HUVEC adhesion induced by LPS was reduced by the intravenous delivery of miPSCs or miPSC-CM. (b) Similarly, the D-HL-60 cell/HUVEC transmigration assay showed that D-HL-60 cell TEM significantly increased after administering LPS and was reduced by the intravenous delivery of miPSCs or miPSC-CM. (c) Western blot assays showed that the expression of VCAM-1 and ICAM-1 in HUVECs significantly increased after administering LPS and was reduced by the intravenous delivery of miPSCs or miPSC-CM. (d) Western blot assays showed that the expression of VLA-4 in D-HL-60 cells significantly increased after administering LPS and was reduced by the intravenous delivery of miPSCs or miPSC-CM. However, the increased expression of LFA-1 in D-HL-60 cells induced by LPS was not reduced by miPSCs or miPSC-CM. Data are presented as the mean ± SD. * p < 0.05, compared to control mice; ** p < 0.05, compared to LPS-induced ALI mice; n = 6 per group.

Fig 5: Western blot assays confirmed that LPS increased the expression of VCAM-1, VLA-4, ICAM-1, and LFA-1 in lung tissue. Administration of miPSCs or miPSC-CM reduced the expression of VCAM-1 and VLA-4 in LPS-induced ALI mice. However, the expression of ICAM-1 in LPS-induced ALI mice could not be reduced by miPSCs. Data are presented as the mean ± SD. * p < 0.05, compared to control mice; ** p < 0.05, compared to LPS-induced ALI mice; n = 6 per group.