Fig. 5

TiO₂ NPs-induced EVs facilitated vascular leakiness and angiogenesis. a EVs was internalized into HUVECs. Time-dependent increase of fluorescence signal intensity was observed after incubation PKH67 labeled EVs with HUVECs. Scale bars = 10 μm. Nuclei, Blue (DAPI); PKH67 labeled EVs, Green; Phalloidin, Red. b Increased fluorescence intensity of FITC-dextran was observed following exposure of HUVECs to EVs for 1 h. Data are shown as mean ± SD (n = 3). c Cell-Tracker Green labeled MDA-MB-231 cells migrated through HUVECs monolayers were observed under fluorescent microscopy following exposure of HUVECs to EVs for 1 h. Scale bars = 50 μm. d PBS-EVs or TiO₂-EVs exposure on HUVECs monolayers resulted in the disruption of VE-cadherin and ZO-1. Scale bars = 10 μm. Nuclei, Blue (DAPI); VE-cadherin or ZO-1, Red. (e) Western blotting analysis of VE-cadherin and ZO-1 in HUVECs that were treated with PBS-EVs or TiO₂-EVs. f The proliferative capacity induced by PBS-EVs or TiO₂-EVs using CCK8 assay. Data are shown as mean ± SD (n = 3). g, h Effects of PBS-EVs or TiO₂-EVs on tube formation ability of HUVECs by tube formation assay. Scale bar = 100 µm. Data are shown as mean ± SD (n = 3). i, j Effects of PBS-EVs or TiO₂-EVs on vascular outgrowth of aortic rings which was quantified by counting all sprouts from one ring. Scale bar = 20 µm. Data are shown as mean ± SD (n = 3). Statistical analysis in (b, f, h and j) was measured by one-way ANOVA