Fig 1: AOPPs accumulation was accompanied by the bone-fat imbalance during skeletal aging. A Serum AOPPs levels in young mice (3-month-old) and aged mice (18-month-old) (n = 6). B AOPPs relative levels in bone marrow from young and aged mice (n = 6). C Serum ALP level (n = 6). D Serum FABP4 level (n = 6). E, F HE staining of bone trabeculae and adipocytes in distal femora, and the area of fat vacuoles was quantified. Scale bars = 100 μm. G, H IHC staining for OCN and number of osteoblasts in distal femoral trabeculas. Scale bars = 20 μm. I, J IHC staining for FABP4 and number of adipocytes. Scale bars = 20 μm. K, L Micro-CT scanning of osmium tetroxide-stained tibia and MAT content below the growth plate (n = 3). M, N The protein and mRNA expression of OCN and PPARγ in proximal tibial metaphysis. O Representative micro-CT images of distal femora from young and aged mice (n = 6) (Scale bars = 1 mm). P-V Quantitative trabecular and cortical bone parameters data were displayed in Bone mineral density (BMD), Trabecular Bone Volume/Total Volume (Tb.BV/TV), Trabecular Number (Tb.N), Trabecular thickness (Tb.Th), Trabecular Separation (Tb.Sp), Cortical Thickness (Ct.Th) and Cortical Bone Area (Ct.BArea) (n = 6). W-Y The correlation analysis of serum AOPPs and BMD, ALP, FABP4. Data were shown as mean ± SD. ∗P < 0.05, ∗∗P < 0.01 vs. young mice group (Student's t test).
Fig 2: Chronic AOPPs loading lead to the bone-fat imbalance in young mice. A Serum AOPPs levels in 6-month-old mice and AOPPs-intervened mice (n = 6). B AOPPs relative levels in bone marrow from 6-month-old mice and AOPPs-intervened mice (n = 6). C Serum ALP level (n = 6). D Serum FABP4 level (n = 6). E, F HE staining of bone trabeculae and adipocytes in distal femora, and the area of fat vacuoles was quantified. Scale bars = 100 μm. G, H IHC staining for OCN and number of osteoblasts in distal femoral trabeculae. Scale bars = 20 μm. I, J IHC staining for FABP4 and number of adipocytes. Scale bars = 20 μm. K, L Micro-CT scanning of osmium tetroxide-stained tibia and MAT content below the growth plate (n = 3). M, N The protein and mRNA expression of OCN and PPARγ in proximal tibial metaphysis. O Representative micro-CT images of distal femora from AOPPs-intervened mice (n = 6) (Scale bars = 1 mm). P-V Quantitative trabecular and cortical bone parameters data were displayed in BMD, Tb.BV/TV, Tb.N, Tb.Th, Tb.Sp, Ct.Th and Ct.BArea (n = 6). Data were presented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01 vs. PBS group (one-way ANOVA).
Fig 3: Decreased AOPPs level by antioxidant leaded to higher bone formation and lower marrow adiposity in aged mice. A Serum AOPPs levels in aged and NAC-treated aged mice (n = 6). B AOPPs relative levels in bone marrow from aged and NAC-treated aged mice (n = 6). C Serum ALP level (n = 6). D Serum FABP4 level (n = 6). E, F HE staining of bone trabeculae and adipocytes in distal femora, and the area of fat vacuoles was quantified. Scale bars = 100 μm. G, H IHC staining for OCN and number of osteoblasts in distal femoral trabeculas. Scale bars = 20 μm. I, J IHC staining for FABP4 and number of adipocytes. Scale bars = 20 μm. K, L Micro-CT scanning of osmium tetroxide-stained tibia and MAT content below the growth plate (n = 3). M, N The protein and mRNA expression of OCN and PPARγ in proximal tibial metaphysis. O Representative micro-CT images of distal femora from aged and NAC-treated aged mice (n = 6) (Scale bars = 1 mm). P-V Quantitative trabecular and cortical bone parameters data were displayed in BMD, Tb.BV/TV, Tb.N, Tb.Th, Tb.Sp, Ct.Th and Ct.BArea (n = 6). Data were shown as mean ± SD. ∗P < 0.05, ∗∗P < 0.01 vs. aged mice group (Student's t test).
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