Fig. 7.

Bone microenvironment changes in macrophage-depleted mice. (A) TUNEL staining on tibial sections in mice (16 wk old, female) treated with clodronate liposomes following the regimen in Fig 5A. (B) TUNEL staining on tibial sections in mice (16 wk old, female) treated with the regimen in Fig 2A. TUNEL⁺ cells were enumerated. n = 7 per group. (C–E) Mice (16 wk old, female) were treated with clodronate liposomes; three consecutive injections were followed by booster injections (every third day, 10 μL/g for 3 wk and 6 μL/g for 1 wk). (C) Flow cytometric analysis of whole marrow cells using anti-CD163 antibody. (D) Genes related to M1/M2 macrophage were analyzed by real-time PCR from whole marrow mRNA. (E) Protein levels of TGF-β1 in bone marrow. **P < 0.01 versus vehicle. (F–H) Mice (16 wk old, female) were treated with clodronate liposomes; three consecutive injections were followed by booster injections (every third day, 10 μL/g for 3 wk and 6 μL/g for 1 wk). Four weeks of intermittent PTH (50 μg/kg) were started after the initial 3 d. Whole marrow mRNA was analyzed by real-time PCR using specific primers for (F) Tgfb1, (G) Wnt3a, and (H) Wnt10b. *P < 0.05; **P < 0.01 versus vehicle-treated PBS; ^#P < 0.05 versus vehicle-treated CLOD. Data are mean ± SEM of two independent experiments. n = 6 per group. CLOD, clodronate liposome; PBS, PBS liposome.