Fig. 8: The effect of Cldn11 in LPS-induced bone destruction and bone formation calvarial mouse models in vivo.

a–d To evaluate the effect of Cldn11 on bone loss in vivo, LPS (500 μg/mouse) or an equal volume of PBS was injected subcutaneously into the calvaria of 5-week-old ICR mice. Recombinant protein of Cldn11 (0.6 mg/mouse) was injected subcutaneously daily for 9 days into the calvaria of LPS-treated mice. e, f Additionally, to investigate the effect of Cldn11 on bone formation in vivo, the same dosage of Cldn11 protein (0.6 mg/mouse) was injected subcutaneously daily for 21 days into the calvaria of normal ICR mice. a TRAP staining of the whole calvaria. b, e Measurements to determine the volume at the treated site and to assess bone formation were carefully performed using 3D image calculator software. The degree of bone density was arbitrarily determined in ROI as follows: low-density (threshold 1101–1600, blue), middle-density (threshold 1601–2100, orange), and high-density (threshold >2100, red). c Histological sections of calvarial bones were stained with hematoxylin and eosin (H&E) and TRAP. d The number of OCs per field was quantified using Image-Pro Plus (Ver 4.5) software. f Histological sections of calvarial bones were stained with H&E. The area of new bone formation was measured using histomorphometric analysis. Data are presented as the mean ± SD. n = 5–6 mice per group. *P < 0.05; **P < 0.01 vs. the control group, ^#P < 0.05; ^###P < 0.001 vs. the LPS group