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. 2020 Mar 4;10(9):3925–3938. doi: 10.7150/thno.41378

Figure 6.

Figure 6

Apilimod treatment improves calvarial defect repair and BMMSC-mediated bone formation. (A) Micro-CT three-dimensional reconstruction of calvarial bone repair in WT mice after treatment with vehicle or apilimod. Red dotted lines indicate the periphery of the defect site. n = 5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). (B) Defect area coverage (%) was calculated via micro-CT images. n = 5 per group. (C-E) Micro-CT analysis including (C) bone volume (BV), (D) bone volume/tissue volume (BV/TV), and (E) healing score for the extent of bony bridging and bone union. n = 5 per group. (F) The H&E staining of calvarial bone defects. Scale bar, 100 µm. (G, H) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT mice for 8 weeks. The mice were given vehicle or apilimod. (G) New bone formation was detected with H&E staining. n = 5 per group. Scale bar, 50 µm. (H) Quantitative analysis of new bone volume. n = 5 per group. (I-K) Bone histomorphometric measurements among each group, including (I) osteoblast number (N.Ob), (J) osteoblast number per bone surface (N.Ob/BS), and (K) osteocyte number (N.Ot). (L, M) Representative images (L) and quantification (M) of immunohistochemical staining of OCN in implants. n = 5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; OCN, osteocalcin; WT, wild-type. Results are shown as mean ± S.D. *p<0.05, **p<0.01, ***p<0.001.