Fig 1: The ALN-functionalized nanosystems demonstrate a dual regulatory capability by promoting the osteogenic differentiation of BMSCs while concurrently inhibiting osteoclastogenesis in BMMs. Representative images of Alizarin Red and ALP staining for BMSCs, and TRAP staining for BMMs (scale bar = 50 μm) (A). The Alizarin Red-stained (B) and ALP-stained (C) areas of cells in each group were quantified using ImageJ software. Statistical analysis of TRAP-positive cell counts in TRAP-stained samples (D). **P < 0.01 vs Control. Data are presented as mean ± SD (n = 3). Western blotting analysis revealed that the ALN-functionalized nanosystems significantly enhanced the expression levels of Collagen I (E) and Osteocalcin (F) in BMSCs. The ALN-functionalized nanosystems significantly inhibited the expression levels of NFATc1 (G) and c-Fos (H) in BMMs, as demonstrated by Western blotting analysis. *P < 0.05, **P < 0.01 vs Control. Data are presented as mean ± SD (n = 3).
Fig 2: The RPT/ALN-PLGA-PEG nanosystems exhibit a rapid and early ability to alleviate inflammation, promote osteogenesis, inhibit bone resorption, and enhance bone mineral density. In the RPT/ALN-PLGA-PEG group, CRP (A) and ESR (B) levels significantly decreased and approached normal values within 2 weeks. BMD returned to normal values within 1 week (C). BALP levels exceeded normal values at 4 weeks (D), while P1NP levels surpassed normal values as early as 2 weeks (E). CTX-I (F) and TRAP-5b (G) levels rapidly declined, reaching near-normal levels within 2 weeks. *P < 0.05, **P < 0.01 vs normal; #P < 0.05, ##P < 0.01 vs TB. Data are presented as mean ± SD (n = 6).
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