Journal: Bioactive Materials

Article Title: Sulfated polysaccharide prevents senescent adipocyte-driven osteonecrosis by stem cell fate reprogramming

doi: 10.1016/j.bioactmat.2025.11.039

Figure Lengend Snippet: Continuous intraosseous administration of SCS prevents glucocorticoid-induced bone degeneration. ( A ) Schematic illustration of the glucocorticoid (GC; MPS)-induced bone deterioration and intraosseous SCS treatment. ( B-D ) Representative H&E staining images of the femur at 6 weeks (B). Magnified views of the cortical bone and trabecular bone in the marrow cavity are shown on the right. Solid arrows indicate normal osteocytes, while hollow arrows indicate empty osteocyte lacunae. Quantification of empty lacunae ratios in cortical bone (C) and trabecular bone (D). n = 6 biological replicates. (Scale bars, 500 μm and 25 μm) ( E-H ) Representative immunofluorescence staining of OPN + mature osteoblasts, osteolectin + osteoprogenitors, and VE-cadherin + endothelial cells (ECs) in femur at 6 weeks (E), and corresponding quantifications (F–H). n = 6 biological replicates. (Scale bars, 100 μm and 20 μm) ( I and J ) Representative flow cytometry plots of capillary subtypes in the femur (I), with quantification of CD45 − Ter119 − CD31 hi Emcn hi ECs (J). n = 6 biological replicates. ( K and L ) Flow cytometry plots showing Sca-1 hi CD31 hi arteriolar ECs (K), and corresponding quantification (L). n = 6 biological replicates. ( M and N ) Representative micro-CT 3D images of the femur (M). Quantitative analysis of percent bone volume (BV/TV) (N). n = 6 biological replicates. (Scale bars, 1.5 mm, 600 μm and 545 μm) ( O and P ) ELISA analysis of VEGF (O) and PDGF-BB (P) levels in bone marrow supernatant and peripheral serum from PBS- and SCS-treated groups at week 6. n = 6 biological replicates. ( Q ) ELISA quantification of the osteogenic factor osteocalcin in peripheral serum at week 6. n = 6 biological replicates. Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant. Statistical significance was determined using one-way ANOVA with Tukey's post hoc test ( C, D, F, G, H, J, L, N, O, P and Q ).

Article Snippet: Serum concentrations of the osteogenic marker osteocalcin (NOVUS, NBP2-68151) were also measured.

Techniques: Staining, Immunofluorescence, Flow Cytometry, Micro-CT, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Sulfated polysaccharide prevents senescent adipocyte-driven osteonecrosis by stem cell fate reprogramming

doi: 10.1016/j.bioactmat.2025.11.039

Figure Lengend Snippet: SCS targets downstream senescent lineage commitment of bone marrow MSCs to mitigate GC-induced bone deterioration. ( A ) Schematic diagram illustrating the experimental design: CD45 − Ter119 − CD31 − LepR + MSCs isolated from mice co-treated with SCS and MPS for 7 days were subjected to in vitro lineage-competitive differentiation, followed by DEX-induced senescence in lineage-mixed cells. These cells were then adoptively transplanted into healthy bone marrow cavity to assess bone deterioration development. ( B ) Representative H&E-stained images of the femur 12 weeks after adoptive transfer. PBS-DEX group: LepR + MSCs from PBS and MPS co-treated mice subjected to in vitro lineage differentiation and DEX-induced senescence, followed by transplantation. SCS-DEX group: LepR + MSCs from SCS and MPS co-treated mice processed similarly. PBS group: solvent control without cell transplantation. Solid arrows indicate intact osteocytes; hollow arrows indicate empty lacunae. (Scale bars, 250 μm and 25 μm) ( C – E ) Quantitative analysis of marrow hypertrophic adipocyte diameter (C), proportion of empty osteocyte lacunae in trabecular bone (D), and adipocyte number (E) in the metaphysis 12 weeks post-transplantation. n = 19 biological replicates (C), n = 6 biological replicates (D), n = 8 biological replicates (E). ( F ) Quantification of empty lacunae in epiphysis at 12 weeks post-transplantation. n = 6 biological replicates. ( G – I ) Representative flow cytometry plots of capillary ECs subtypes in the femur at 12 weeks (G), with quantification of CD45 − Ter119 − CD31 hi Emcn hi ECs (H) and CD45 − Ter119 − CD31 lo Emcn lo ECs (I). n = 6 biological replicates. ( J and K ) Representative flow cytometry plots (J) and corresponding quantification (K) of CD45 − Ter119 − Sca-1 hi CD31 hi arteriolar ECs in the femur at 12 weeks post-transplantation. n = 6 biological replicates. ( L ) Representative micro-CT images of the femur at 12 weeks post-transplantation across different treatment groups. (Scale bars, 1.5 mm and 500 μm) ( M – P ) Quantitative analysis of bone parameters in the metaphysis: bone mineral density (BMD) (M), percent bone volume (BV/TV) (N), trabecular separation (Tb.Sp) (O), and trabecular number (Tb.N) (P). n = 6 biological replicates. ( Q ) Serum ELISA analysis of the osteogenic marker osteocalcin at 12 weeks post-transplantation. n = 6 biological replicates. ( R and S ) ELISA analysis of PDGF-BB (R) and VEGF (S) in both bone marrow supernatant and peripheral serum at 12 weeks post-transplantation. n = 6 biological replicates. Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant. Statistical significance was determined using one-way ANOVA with Tukey's post hoc test ( C, D, E, F, H, I, K, M, N, O, P, Q, R and S ).

Article Snippet: Serum concentrations of the osteogenic marker osteocalcin (NOVUS, NBP2-68151) were also measured.

Techniques: Isolation, In Vitro, Staining, Adoptive Transfer Assay, Transplantation Assay, Solvent, Control, Flow Cytometry, Micro-CT, Enzyme-linked Immunosorbent Assay, Marker

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 reduces Sema4D levels to elevate cortical strength in mice with type 2 diabetes mellitus (T2DM). (A) Schedule of enzyme-linked immunosorbent assay (ELISA) and the corresponding groups. (B) ELISA to determine the levels of ALP, Runx-2, OCN, OPN, and Sema4D in control mice, mice with T2DM, mice with T2DM treated with metformin (Met)/exendin-4 (Ex-4), and GLP-1R knockdown T2DM mice treated with Ex-4. n=6 mice per group. ALP: Alkaline Phosphatase; Runx-2: Runt-related transcription factor 2; OCN: Osteocalcin; OPN: Osteopontin; Sema4D: Semaphorin 4D. (C) Schedule of micro-CT to determine changes in trabecular bone. (D) Micro-CT images representing trabecular formation in control mice, mice with T2DM, mice with T2DM treated with Ex-4, mice with T2DM treated with anti-Sema4D, and mice with T2DM treated with a combination of Ex-4 and anti-Sema4D within a region of interest (ROI). n=6 mice per group. (E) Quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 mice per group. (F, G) Micro-CT images representing cortical bone formation in control mice, mice with T2DM, mice with T2DM treated with Ex-4, mice with T2DM treated with anti-Sema4D, and mice with T2DM treated with a combination of Ex-4 and anti-Sema4D within an ROI, and quantitative analysis of micro-CT images. n=6 mice per group. (H) Effects of exendin-4 or anti-Sema4D on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 mice per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: ELISA was conducted following the protocols of the Mouse soluble Cluster of differentiation 100 (sCD100) ELISA Kit (EM1344, FineTest), Mouse Alkaline Phosphatase (ALP) ELISA Kit (EM0828, FineTest), Mouse Runt Related Transcription Factor 2 (RUNX2) ELISA Kit (EM1334, FineTest), Mouse Osteocalcin (OC/BGP) ELISA Kit (E-EL-M0864, Elabscience), Mouse Osteopontin (OPN) ELISA Kit (E-EL-M3030, Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Knockdown, Micro-CT, Standard Deviation

Journal: bioRxiv

Article Title: Interleukin-3 as a Potential Bone Anabolic Agent in treating Postmenopausal Osteoporosis

doi: 10.1101/2025.07.12.664485

Figure Lengend Snippet: IL-3 alters OVX-induced changes in osteoclasts- and osteoblasts-specific gene expression. Marrow free bones from IL-3 treated animals from both Prophylactic and Therapeutic treatment strategy, as indicated, were subjected to mRNA expression profile and ( A, B ) Osteoclast-specific markers: integrinβ3, Dc-stamp, and Nfatc1 , Ctsk, Tnfrsf11a(RANK), Mmp9, Trap, and Ctr. (B, D) Osteoblast-specific markers: Runx2, Osx, Opn, Alp, Bmp2, Bmp4, Ocn, Bmp6, and Col1 were assessed by qPCR. Serum ( E, G ) CTX and ( F, H ) OCN were also measured by ELISA in both the strategies. Data is presented as mean ± SEM with n=3 mice/group for qPCR, and n=8 mice/group for ELISA.

Article Snippet: The serum level of CTX-I (CUSABIO; CSB-E12782m) and OCN (CUSABIO ; CSB-E06917m) was assessed by ELISA according to manufacturers’ instructions.

Techniques: Gene Expression, Expressing, Enzyme-linked Immunosorbent Assay