Figure 5.

C5ar1 deficiency partially prevented bone loss induced by chemotherapy

(A) Quantitative parameters and 3D representation of the trabecular compartment of tibiae measured by μ-CT from wild-type (WT), WT/DoxoR, C5aR1^−/−, and C5aR1^−/−/DoxoR experimental groups. n = 9–13. (B) Quantification of the number of osteoblasts per bone surface (osteoblasts/mm) and osteocytes per cortical bone area (osteocytes/mm²) from H&E-stained sections of tibiae. The number of osteoclasts per trabecular bone surface (osteoclasts/mm) was quantified from TRAP staining of tibiae. n = 4–6. (C) qRT-PCR of osteoblastic (Runx2, Osx, and Col1a1) and osteoclastic (Nfatc1, Mmp9, Ctsk, and Trap) markers. RNA isolated from the tibiae and femurs of WT, WT DoxoR, C5aR1^−/−, and C5aR1^−/−/DoxoR experimental groups. n = 5–9. (D) Chemotaxis of primary monocytes isolated from WT and C5aR1^−/− mice toward medium without FBS, medium without FBS + 0.1 μg/mL of C5a (C5a). Migration of WT monocytes pre-treated for 30 min with 0.9 μg/mL PMX53 toward medium without FBS + 0.1 μg/mL C5a was also evaluated n = 4–6. (E) qRT-PCR of osteoclastic gene expression (Csf1r, Rank, Nfatc1, Mmp9, Ctsk, and Trap) from primary WT and C5aR1^−/− of primary WT and C5aR1^−/− monocytes. n = 8–11. Data are shown as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (A–D) One-way ANOVA. (E) Student’s t test.