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. 2021 Mar 15;162(7):2070–2086. doi: 10.1097/j.pain.0000000000002207

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Copyright © 2021 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the International Association for the Study of Pain.

This is an open access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.

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Figure 2. — Study rationales—behavior profiles and DRG levels. (A) On plantar incision (INC), pain-related behavior profiles of nonevoked (NEP) and evoked (mechanical and heat) pain-related behaviors were evaluated in an independent cohort (cohort 1, green, INC N = 8, Sham N = 8). INC-induced NEP, as well as mechanical and heat hypersensitivity started in the acute phase (4∼6 hours) after surgery and lasted up to postoperative day (POD) 3 (mechanical and heat hypersensitivity) and up to POD 6 (NEP), respectively. At POD 1, 2, and 3, no significant differences were found for either pain-related behavior. Mice were randomly assigned to experimental groups (INC and Sham), and experiments were performed in a balanced study design. Data are expressed as change in % (mean ± SEM). Statistics were performed by 2-way ANOVA followed by Dunnett's post-hoc test. P-Values: ***P < 0.001 vs baseline (BL), ^†††P < 0.001 vs Sham. (B) The contribution of DRG levels to the sciatic nerve was determined by retrograde tracing (True Blue dye) in a separate cohort of male naïve mice of our local C57BL6/J colony (cohort 2, grey, naïve N = 8). Representative images of the ipsilateral lumbar DRG of levels L3 to L6. True Blue–positive neurons were counted 7 days after tracer injection and are significantly increased in ipsilateral DRG L4, and L5 compared with L3 and L6. The results are expressed as the absolute number of True Blue–positive and negative neurons, and the percentage of neurons in total. Scale bar = 200 µm. Statistics: Fisher's exact test (contingency). P-Values: ***P < 0.001. (C) Determination of behavior-based inclusion criterion for cohorts 3 and 4 (cohort 3, blue, INC N = 15, Sham N = 15; cohort 4, red, INC N = 12, and Sham N = 12). At POD 1, INC-induced significant NEP in both cohorts. The results are expressed as mean (ratio incised/nonincised paw) ± SEM and single values for each mice (open circles). P-Values: ***P < 0.001 vs baseline (BL), ^†††P < 0.001 vs Sham by 2-way ANOVA followed by Dunnett's post-hoc test. ANOVA, analysis of variance; DRG, dorsal root ganglia.