Figure 3.

miR-672-5p Treatment Has an Anabolic Effect and Promotes Bone Formation

(A) Immunofluorescence assay in decalcified bone sections showed that miR-672-5p overexpression promoted Runx2 expression (red) and suppressed Smurf1 expression (green) as compared with the PBS-treated group (n = 3, magnification 20×). (B and C) Representative photomicrographs of Goldner’s trichrome (GT) staining (B) of distal femurs from different groups and histomorphometric quantification; BS/TV (C) using the Bioquant software. Data are mean ± SE. *p < 0.05 compared with the PBS-treated group, or as shown ˆp < 0.05; (n = 3, magnification 20×). (D) Osteogenic marker; serum P1NP concentration of different groups by ELISA. Data are mean ± SE. **p < 0.01 compared with the PBS-treated group, or as shown ˆˆˆp < 0.001; (n = 6). (E) Representative photomicrographs of the calcein double labeling in the femur (n = 3, magnification 20×). (F) Quantification of data showing mineralizing surface normalized with bone surface (MS/BS), mineral apposition rate (MAR), and rate of bone formation, surface referent (BFR/BS). Data are mean ± SE. *p < 0.05, **p < 0.01 and ***p < 0.001 compared with the PBS-treated group, or as shown ˆˆp < 0.01 and ˆˆˆp < 0.001. See also Figures S2A–S2C for histological examination of the femur metaphysis for osteoclast numbers and bone resorption surface estimates for in vivo experiments.