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. 2022 May 31;79(6):330. doi: 10.1007/s00018-022-04335-w

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© The Author(s) 2022

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

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Fig. 6 — Nociceptors enhance osteoblast functioning and inhibit osteoclast activity. Quantification of Osteoimage fluorescent hydroxyapatite staining (a) (n = 7–8 per group). Representative 20 × images frsom MC3T3-E1 cultured in osteogenic differentiation media with trigeminal ganglia from Nav1.8-Cre [+ TG (cre)] or Nav1.8-DTA [+ TG (DTA)] mice for Osteoimage (b). Quantification of Alizarin Red calcium staining (c) (n = 5–7 per group). Representative 20 × images from MC3T3-E1 cultured in osteogenic differentiation media with trigeminal ganglia from Nav1.8-Cre (+ TG (cre)) or Nav1.8-DTA (+ TG (DTA)) mice for Alizarin Red (d). Data are presented as either relative fluorescence or absorbance as percent of control (i.e., MC3T3-E1 cells cultured alone). Quantification of alkaline phosphatase activity and secreted osteocalcin of MC3T3-E1 cells after co-culture with Nav1.8-Cre or Nav1.8-DTA TG neurons (e and f, respectively) (n = 6 per group and n = 14–16 per group, respectively). Data are presented as fluorescent (ALP activity) and absorbance (osteocalcin expression) readings normalized to control. Quantification of RAW264.7 bone resorption activity, presented as fluorescent reading normalized to control (g) (n = 6–7 per group). Quantification of multinucleated osteoclasts, represented as total number of multinucleated cells per field of view (h) (n = 8 per group). Statistical analysis was performed using a two-way ANOVA. All graphs are shown as mean ± SEM