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. 2020 May 8;10:7815. doi: 10.1038/s41598-020-64783-x

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© The Author(s) 2020

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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Development of undulations and varicosities. (A) Effect of minocycline on undulations developing into varicosities. Measured over 30 days. Significant differences were observed in the percentage of undulations that developed to varicosities between the Sham-vehicle and TBI-vehicle (p = 0.02) groups, and TBI-72-hr-minocycline and Sham-vehicle groups (p = 0.01). Data are presented as mean ± SD. Number of mice imaged is listed in Table 1. *p < 0.05 vs. Sham-vehicle, one-way ANOVA was performed to test the percentage of undulations that developed to varicosities (p = 0.008, ANOVA power = 0.96). Tukey’s test was performed for pairwise comparisons. (B) Axons with varicosities at 5 time points after experimental TBI. The percentage of axons with varicosities was measured at each time point. Two-way ANOVA shows that main effects of treatment (F=18.82, p < 0.00001) and time (F = 10.64, p < 0.00001) are significant. Tukey’s test was performed for pairwise comparisons. The Tukey test results show a significant difference between TBI-vehicle (p < 0.0001), TBI-45-min minocycline treated mice (p < 0.0001), TBI-72-hr minocycline treated mice (p < 0.0001) as compared with Sham-vehicle mice. All TBI groups exhibited axons with varicosities, independent of treatment. TBI-vehicle treated mice had significantly more axons with varicosities on Day 3 (p = 0.011), Day 7 (p = 0.002), and Day 14 (p = 0.01) compared to the Sham-vehicle mice. The percentage of axons with varicosities in TBI-45-min minocycline treated mice (mino in legend) were not significantly different than the Sham-vehicle mice at any time point. The TBI-45-min minocycline treated mice had a non-significant, smaller percentage of axons with varicosities than the TBI-vehicle mice. The TBI-72-hr minocycline treated mice had significantly more axons with varicosities on Day 3 (p = 0.018) and Day 7 (p = 0.029) as compared to the Sham-vehicle mice. By Day 14, the TBI-72-hr minocycline treated mice had a non-significant, smaller percentage of axons with varicosities than the TBI-vehicle treated animals (p = 0.188). Data are presented as mean ± SD. *p < 0.05, **p < 0.01.