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. 2017 Aug 15;7:8180. doi: 10.1038/s41598-017-08250-0

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© The Author(s) 2017

Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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e-sEVs increase capillary density and protect muscle from ischemic damage. (A) Representative hematoxylin-eosin images of gastrocnemius muscles of non-ischemic (Normal hind limb) and ischemic hind limb of Vehicle, e-sEVs and i-sEVs treated animals (Original Magnification: ×200; scale bar: 50 µm), (upper panels). Representative immunofluorescence images (confocal microscopy) of capillary density staining. An anti-CD31 mAb was used to analyze gastrocnemius muscles of non-ischemic (Normal hind limb) and ischemic hind limbs as indicated (Original Magnification: ×400; scale bar: 25 µm) (lower panels). (B) Quantitative analysis of muscle damage areas in hematoxilin-eosin stained gastrocnemius muscles of normal and ischemic hind limbs at day 7 after surgery. e-sEVs significantly reduce muscle damage compared to Vehicle. Data are expressed as mean ± SEM; *p < 0.05 e-sEVs vs Vehicle, (One-way ANOVA - Newman-Keuls Multiple Comparison Test), (left panel) (n = 10). Quantitative analysis of capillary density of normal and ischemic hind limbs at day 7 after surgery indicates that e-sEVs significantly increase capillary density compared to Vehicle. Data are expressed as mean ± SEM; *p < 0.05, e-sEVs vs Vehicle, (One-Way ANOVA - Newman-Keuls Multiple Comparison Test), (right panel).