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. 2018 Jan 11;8:480. doi: 10.1038/s41598-017-18867-w

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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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Microglia activation in injured spinal cord Samples of spinal cord tissue containing the injury epicenter (a) were analyzed by flow cytometry for microglia activation state phenotypes. Interestingly, the total number of cells positive for markers associated with pro-inflammatory, M1 microglia (CD32 and CD86) were significantly decreased after treatment with MSCEv^wt and MSCEv⁺ (***p < 0.001, **p < 0.01, respectively)(b). Additionally, lumbar sections of spinal cord were analyzed, revealing the ratio of positive cells in injured sections of spinal cord and distal lumbar sections of spinal cord. Cells positive for M1 associated markers CD32 and CD86 are increased after SCI but treatment with either MSCEv^wt or MSCEv⁺ decrease levels of these cells to near sham values. Cells positive for M2 associated markers CD200R, CD163 and RT1B are significantly decreased (**p < 0.01, *p < 0.05, *p < 0.05, respectively) with treatment of either MSCEv^wt or MSCEv⁺ after SCI. Overall, there is a significant reduction in the number of activated microglia after treatment with either MSCEv^wt or MSCEv⁺, indicating that both EVs are effective in reducing inflammation in the injury epicenter as early as 14 days post-injury (c). Sham + Vehicle, n = 2, SCI + Vehicle, n = 2, SCI + MSCEv^wt, n = 3, SCI + MSCEv⁺, n = 3.