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. 2022 Dec 17;26:83. doi: 10.1186/s40824-022-00325-y

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Stimulated angiogenesis mediated by ADSC-NVs in vitro. The dose-effect of ADSC-NVs and ADSC-EVs on cell migration (C) and tube formation (D) abilities of HUVECs was confirmed. Considering the possible side effects and enormous economic cost of high concentrations [28], different doses of ADSC-EVs and ADSC-NVs (ranging from 5 to 80 μg/mL) were assessed. Transwell assay was conducted following the shown schematic diagram (A). Briefly, HUVECs on the upper side of the transwell membrane were treated with different dose of ADSC-NVs or ADSC-EVs (0, 5, 20 and 80 μg/mL) in the lower side for 24 h. The cells migrated to the bottom of the membrane were fixed with 4% paraformaldehyde and stained with crystal violet. C Representative images of transwell assay with ADSC-EVs or ADSC-NVs treatment. Scale bar: 10 μm. B Quantitative analysis of the migrated cells (n = 4 per group). D Formation of tube-like structures by HUVECs-GFP after incubation with different doses of ADSC-EVs or ADSC-NVs (0, 5, 20, and 80 μg/mL) for 6 h. Scale bar: 100 μm. E Quantitative analysis of tube-like structures formation (n = 3 per group). F mRNA expression of angiopoietin-1 (Ang-1) and vascular endothelial growth factor (VEGF) in HUVECs treated with ADSC-NVs or ADSC-EVs was confirmed using qRT-PCR (n = 3 per group). G Expression levels of Ang-1 and VEGF proteins in HUVECs after ADSC-NVs or ADSC-EVs stimulation. ns, no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001