Fig. 4
From: Inhibition of overactive TGF-β attenuates progression of heterotopic ossification in mice
Systemic injection of TGF-β neutralizing antibody reduces ectopic bone formation and type H vessel formation. a–c Mice were treated with 5 mg/kg body weight of the TGF-β neutralizing antibody 1D11 three times a week for 3 weeks from 1 day (D1), 3 weeks (W3), 6 weeks (W6), and 12 weeks (W12) after ATP and analyzed 15 weeks after ATP or sham surgery. a Micro CT images of the Achilles tendon (sagittal view) of mice and b quantitative analysis of bone volume of heterotopic bone determined by μCT analysis. Scar bar, 2 mm. c H&E staining of ectopic bone of Achilles tendons. Scale bar, 50 μm. d–l Mice were treated with 5 mg/kg body weight of 1D11 three times a week for 3 weeks from 1 day (D1), 3 weeks (W3), and 6 weeks (W6) after ATP and analyzed 9 weeks after ATP or sham surgery. d Immunostaining and e quantification of pSmad2/3+ cells in ectopic bone marrow. Scale bar, 50 μm. f Active TGF-β in serum determined by ELISA. g Nestin+ (red) cells in the ectopic bone marrow and h quantification. Scale bar, 50 μm. Blue indicates DAPI staining of nuclei. i CD31+ (red) and Emcn+ (green) cells in the ectopic bone marrow. j Quantification of the fold change of Type H vessels in 1D11-treated ATP mice normalized to that of Sham mice. Scale bar, 100 μm. Yellow indicates type H vessels. k Immunostaining and l quantification of Ocn+ cells of in ectopic bone marrow. Red arrow shows Ocn+ cells. Scale bar, 50 μm. All data are shown as the mean ± s.d. n = 8 per group. *p < 0.05 as determined by one-way ANOVA
