Figure 5.

Therapeutic Inhibition of miR-208a-3p with Bone-Targeting Delivery System Counteracts Decrease in Bone Formation of Hindlimb-Unloaded Mice

(A) qPCR analysis of miR-208a-3p levels in HLU bone and other tissues from C57BL/6J mice. (B) A schematic diagram illustrating experimental design. (C) Relative expression of miR-208a-3p and ACVR1 in femurs collected from baseline, control, HLU, HLU+PBS, HLU+antagomiR-NC, and HLU+antagomiR-208a-3p groups. (D) Representative μCT reconstructive images of distal femurs collected from baseline, control, HLU, HLU+PBS, HLU+antagomiR-NC, and HLU+antagomiR-208a-3p mice. (E) Representative images showing new bone formation assessed by double calcein labeling in mouse femurs collected from baseline, control, HLU, HLU+PBS, HLU+antagomiR-NC, and HLU+antagomiR-208a-3p. Scale bars, 10 μm. (F) BMD and BMC measurements in the femurs collected from baseline, control, HLU, HLU+PBS, HLU+antagomiR-NC, and HLU+antagomiR-208a-3p mice. (G) Trabecular bone parameters of bone volume/tissue volume ratio (BV/TV), trabecular separation (Tb.Sp), trabecular number (TB.N), and trabecular thickness (Tb.Th) in baseline, control, HLU, HLU+PBS, HLU+antagomiR-NC, and HLU+antagomiR-208a-3p mice. (H) Histomorphometric analysis of bone-formation-related parameters BFR and Ob.S/BS in HLU, HLU+PBS, HLU+antagomiR-NC, and HLU+antagomiR-208a-3p mice. (I) qPCR analysis expression of osteogenic genes ALP, Col Ia1, Ocn, and Runx2 in six group. Each experimental group was compared to the others. All data were expressed as means ± SD. Significance is noted at these thresholds: *p < 0.05, **p < 0.01, ***p < 0.001. One-way ANOVA with a post hoc test was performed. Statistical differences between two groups were determined by Student’s t test. n = 8 mice in each group.