Fig. 7: Snhg18 delivery mitigates bone loss in OVX- and disuse-induced osteoporosis.

a Study and treatment methods diagram in OVX-induced osteoporosis model with AAV9 injection (i.v., 4 × 10¹¹ GC). Seven weeks later, mice received subcutaneous calcein and alizarin injections at 6-day intervals for dynamic histomorphometry. b Double calcein/alizarin labeling showing new bone formation rates in femurs, with MAR quantification. Scale bars, 100 µm. n = 6 mice per group. c Representative micro-CT images of the femur and mandible in Sham + NC, OVX + NC, and OVX + Snhg18 groups. Scale bars, 1 mm. d Quantitative analysis of BV/TV and Tb.Th in femur. n = 6, 8, and 8 mice per group. e Von Kossa staining of femur. Scale bars, 1 mm. f Masson’s trichrome staining of the femur in Sham + NC, OVX + NC, and OVX + Snhg18 groups. Scale bars, 100 µm. Experiments were conducted independently 3 times, consistently producing similar results. g Immunofluorescence images and quantification of Yap localization in LepR⁺ MSCs in the femur, with DAPI (blue), Yap (yellow), and LepR (purple) staining, and AAV-driven fluorescence (green). The white arrow points to the nucleus of the AAV-targeted LepR⁺ MSCs. Scale bars, 20 µm. n ≥ 100 cells per group from three independent experiments. h Study and treatment timeline in disuse osteoporosis model with AAV9 injection (i.v., 4 × 10¹¹ GC). Created in BioRender. Tu, D. (2025) https://BioRender.com/kdp734f. i Masson’s trichrome staining of mandible with BV/TV and Tb.Th quantification. Scale bars, 500 µm. n = 6 mice per group. j Immunofluorescence images and quantification of Yap localization in LepR⁺ MSCs in the mandible, with DAPI (blue), Yap (yellow), and LepR (purple) staining, and AAV-driven fluorescence (green). The white arrow points to the nucleus of the AAV-targeted LepR⁺ MSCs. Scale bars, 50 µm. n ≥ 100 cells per group from three independent experiments. Statistical analyses were conducted as follows: One-way ANOVA for (b, d, g, i, j). Data are presented as mean ± SD. MAR mineral apposition rate, BV/TV bone volume per tissue volume, Tb.Th trabecular thickness. Source data and exact p values are provided in the Source data file.