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. 2022 Jan 19;17:457–470. doi: 10.1016/j.bioactmat.2022.01.019

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© 2022 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 3 — Characterization and pro-osteogenic ability of Nell1-modified-EVs (Nell1/EVs). (A) Representative transmission electron microscopy images of the ultrastructure of Nell1/EVs and Ctrl/EVs. Scale bars: 200 nm. (B) Nanoparticle transport analysis of the size distribution and particle concentration of the two types of EVs. (C) Representative immunoblots showing expression of Alix, CD54, Flotillin-1, Annexin V and NELL1. GM130 was used here as a negative marker and ACTIN as a loading control. (D) RT-qPCR analysis of osteoblast gene markers in BMSCs treated with PBS, Nell1/EVs, or Ctrl/EVs at 3 and 6 days. (E) ALP staining images and ALP activity in BMSCs treated with PBS, Nell1/EVs, or Ctrl/EVs on day 14. (F) Alizarin Red-stained images and semi-quantification of BMSCs treated with PBS, Nell1/EVs, or Ctrl/EVs at day 21. *p < 0.05, **p < 0.01, ***p < 0.001, NS, non-significant, via t-test or one-way analysis of variance, n = 3.