Fig. 6: Self-renewable VEGF 165 from HUVEC^vegf165+-laden hydrogel improves HG-induced endothelial cell injuries via protecting mitochondrial permeability.

A HG impairs the viability of HUVECs. OC, osmotic control with 5 mM glucose and 35 mM D-mannitol. n = 3 independent experiments. B Schematic diagram of cell models used to investigate the effects of the supernatant of cell-laden hydrogel platform on HG-induced HUVEC injuries. This figure was created with BioRender.com and has been granted a publication license. C Supernatant of HUVEC^vegf165+-laden hydrogel platform significantly improved the viability of HUVECs under the HG condition. n = 3 independent experiments. D Supernatant of HUVEC^vegf165+-laden hydrogel platform significantly reduced the ROS production of HUVECs under the HG condition. n = 3 independent experiments. E Immunofluorescence (IF) staining assay revealed HG-induced DNA/RNA damages with the IF signals co-localized with mitochondria. The supernatant of HUVEC^vegf165+-laden hydrogel platform ameliorated the DNA/RNA damage. F Quantitative analysis of mitochondrial DNA/RNA damage after different treatments. n = 3 independent experiments. G Supernatant of HUVEC^vegf165+-laden hydrogel platform improved the mitochondrial permeabilization detected with JC-1, a type of mitochondrial membrane potential (MMP) dye. H Quantitative analysis of the fluorescence intensity ratio of JC-1 aggregates to JC-1 monomers for determining MMPs after different treatments. n = 3 independent experiments. I Western blot results indicated that the VEGF 165 in the supernatant of HUVEC^vegf165+-laden hydrogel platform alleviated Bax-elicited mitochondrial perforation and casp-3-activated programmed cell death due to mitochondrial leakage. The results have been confirmed by two independent experiments. The p values in the figures (A, C, D, F, and H) are determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data are presented as mean ± SD. ns, not significant. Source data are provided as a Source Data file.