Fig. 1.

Chronic inflammation augments OC activity and expands Ly6Chi monocyte populations. a Time course of the vaccination model (see the Methods section for details). b Representative 3D modeling from femoral microCT scans of control (cont.) and inflamed (inf.) mice. All microCT scans were performed on the right femur. c MicroCT evaluation of the cortical bone cross-sectional area fraction (Ct.Ar/Tt.Ar) and trabecular bone volume/total volume (BV/TV). d Histomorphometric indices: bone formation rate/bone surface (BFR/BS) and fraction of OCs occupying the cortical perimeter (Oc.Pm/BPm). The full microCT and histomorphometric results are shown in Table S1. e Serum collagen type 1 cross-linked C-telopeptide (CTX-I) levels. f Relative quantities (RQs) of Tnf-α, Rankl, M-csf and Opg transcripts in RNA extracted from the proximal half of the right tibia. a–f Control n = 7, inflamed n = 7; representative results for three independent experiments (Mann–Whitney test and Holm multiplicity correction). g Flow cytometric analysis of leukocytes isolated from the BM and blood of control and inflamed mice. The plots are gated on Lin^-(Thy1^.2^-B220^-Ter119^-Ly6G^-)CD115⁺ monocytes and depict the Ly6C^hiCD11b^hi and Ly6C^hiCD11b^lo subsets. h Ly6C^hiCD11b^hi and Ly6C^hiCD11b^lo cell frequencies and absolute numbers in the BM of control and inflamed mice [absolute number in BM extracted from 1 leg (femur + tibia)]. i Frequency in the blood. Representative results for three independent experiments are presented. Control n = 15, inflamed n = 15 (Mann–Whitney test and Holm multiplicity correction). j Kinetics of Ly6C^hiCD11b^hi and Ly6C^hiCD11b^lo cell accumulation in the BM during the vaccination protocol. Inflamed n = 7 for each time point, Day 16 control n = 6 (Kruskal–Wallis and Dunn post-test all compared to Day 0). Line: median, box: 25^th–75^th percentile, whiskers: range. *P < 0.05, **P < 0.01, ***P < 0.00 1, ****P < 0.000 1. (s.c. – subcutaneous, i.f. – intrafootpad, i.p. – intraperitoneal)