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. 2017 Aug 31;7:10107. doi: 10.1038/s41598-017-10251-y

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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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The role of monocyte/macrophage in BMSC’s antihyperalgesia. Liposome-encapsulated clodronate (Lipo-Clo) was injected i.p. 3 times in three days (10 mg/2 ml/injection/rat). (a,b) Lipo-Clo treatment attenuated BMSC-produced antihyperalgesia (a, pretreatment, b, post-treatment). Compared to Veh-treated rats, Lipo-Clo treatment led to a reduction in EF₅₀. Veh: Phosphate buffered saline-filled liposome. Error bars are 95% confidence intervals. *p < 0.05, **p < 0.01, ***p < 0.001, vs. 14d-TL; #p < 0.01, ###p < 0.001, vs. veh. n = 5-9/Group. (c) RVM MOR mRNA levels were reduced in Lipo-Clo-treated rats (Sample taken at 7d post-Lipo-Clo from BMSC-treated rats). P < 0.05, n = 3. (d) Flow chart of the experiments shown in (e and f). Bottom image shows the injection site in RVM. Coronal brainstem sections were stained with green fluorescent Nissl stain. Arrow indicates the injection needle track and circle shows the site of injection. (e) Injection of MC (Arrow, 10,000 cells/0.5 μl) from BMSC-treated TL animals into the RVM attenuated behavioural hyperalgesia in TL rats, as compared to MC from medium-treated TL rats. ***p < 0.001 vs. Med-MC, n = 6/Group. (f) BMSC induced changes in monocyte phenotype. RT-qPCR was performed on peripheral blood monocytes isolated from TL rats treated with primary or 20P-BMSC or culture medium. Note that four M2 macrophage related markers were upregulated in primary BMSC-treated rats. *p < 0.05, ***p < 0.001, vs. naive; ^#p < 0.05, ^##p < 0.01, vs. medium; ⁺p < 0.05, vs. 20 P, n = 3. Statistics: (a,b,e) Two-way ANOVA followed by post-hoc comparisons with Bonferroni corrections; (c) Unpaired, Two-tailed Student’s t-test; (f) One-way ANOVA followed by post-hoc comparisons with Bonferroni corrections.