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 modulates mesenchymal stem cell lineage bias via activation of the IGF-1/PI3K/Akt/mTOR signaling pathway. ( A ) Quantitative analysis of osteocyte morphology in the trabecular bone matrix of the bone marrow at week 6 after MPS treatment with or without SCS, in the presence of various neutralizing antibodies (NAbs) and antagonistic proteins. ( B ) ELISA analysis of IGF-1 and BMP-2 levels in the femoral bone marrow and peripheral serum at day 7 following SCS treatment under MPS conditions. ( C and D ) Western blot analysis of phospho-PI3K, phospho-Akt, and phospho-mTOR (C), as well as phospho-Smad1/5/8, phospho-ERK, and phospho-p38 (D), in CD45 − Ter119 − CD31 − LepR + MSCs after 15-min stimulation with conditioned medium (CM) derived from bone marrow fluid at day 7 following SCS treatment. ( E – G ) Representative flow cytometry plots (E, F) and quantitative analysis (G) of CD45 − CD31 − Sca-1 + CD24 − adipocyte progenitor cells (APCs), CD45 − CD31 − Sca-1 + CD24 + MSCs (E), and CD45 − CD31 − Sca-1 − PDGFRα + (Pα + ) osteoprogenitor cells (OPCs) (F) from femoral bone marrow at day 14 post-MPS induction with or without combined treatment using SCS and IGF-1 NAb or Noggin. ( H and I ) Representative SA-β-Gal staining images (green) of the femur (H), and corresponding quantification (I), at week 4 following MPS treatment with SCS in combination with IGF-1 NAb or DMH1. Insets show magnified views of bone marrow (BM) and trabecular bone matrix (TBM) regions. (Scale bars, 100 μm and 25 μm) ( J ) qPCR analysis of 12 senescence-associated markers in ex vivo femoral bone tissues at week 4 following MPS treatment with SCS in combination with IGF-1 NAb or DMH1. ( K ) Representative Oil Red O staining images of CD45 − Ter119 − CD31 − LepR + MSCs sorted from femurs at day 7 following MPS treatment with SCS in combination with LY294002 or LDN-193189, after in vitro adipogenic induction. (Scale bars, 50 μm and 25 μm) ( L and M ) γ-H2A.X and telomere-associated DNA damage foci (TAFs) co-localization analysis (L), and corresponding quantification (M), in CD45 − Ter119 − CD31 + arteriolar ECs sorted from femurs at day 28 following MPS treatment with SCS in combination with rapamycin or LDN-193189, using immuno-FISH staining. (Scale bars, 7 μm and 1 μm) ( N and O ) Sequential fluorescent labeling using calcein (N) and quantification of mineral apposition rate (O) in femurs treated with SCS and MPS for 4 weeks, with or without LY294002 and/or GW9662. (Scale bars, 50 μm) ( P ) ELISA analysis of five senescence-associated cytokines in femoral bone marrow at day 28 following MPS treatment with SCS in combination with rapamycin and/or T0070907. ( Q and R ) Representative t-distributed stochastic neighbor embedding (t-SNE) plots (Q) from flow cytometric analysis of CD45 − CD31 − Sca-1 + CD24 − APCs, CD45 − CD31 − Sca-1 + CD24 + MSCs, CD45 − CD31 − Sca-1 − Pα + OPCs, CD45 − Ter119 − CD31 + arteriolar ECs, and CD45 − Ter119 − Emcn + sinusoidal ECs at day 14 following MPS treatment with SCS in combination with IGF-1 and/or rosiglitazone, and quantitative analysis of APCs (R) ( S ) Heatmap showing the fluorescent intensity distribution of Lamin-B1 expression across five cellular subpopulations as identified in the t-SNE clustering plot. ∗ P < 0.05 vs. IgG (empty lacunae); # P < 0.05 vs. IgG (filled lacunae). ∗ P < 0.05 vs. SCS; # P < 0.05 vs. SCS + IGF-1 NAb. 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 an unpaired two-tailed Student's t -test ( B ), or one-way ANOVA with Tukey's post hoc test ( A, G, I, J, O, P and R ).

Article Snippet: Other drugs and compounds used in this study included: GW9662 (MCE, HY-16578; intraperitoneal injection, 1 mg/kg body weight/day, administered continuously for 4 weeks), T0070907 (Selleck, S2871; intraperitoneal injection, 2 mg/kg body weight/day, administered continuously for 4 weeks), rapamycin (MCE, HY-10219; subcutaneous injection, 3 mg/kg body weight/day, administered continuously for 4 weeks), Rosiglitazone (MCE, HY-17386; oral gavage, 3 mg/kg body weight/day, administered continuously for 2 weeks), LY294002 (Selleck, S1105; intraosseous injection, 10 μM, 5 μL per dose per week, administered for 1 or 4 weeks), DMH1 (Selleck, S7146; intraperitoneal injection, 5 mg/kg body weight/day, administered continuously for 4 weeks), Noggin (PeproTech, 250-38; intraosseous injection, 50 ng per dose, twice per week, administered for 2 or 4 weeks), LDN-193189 (Selleck, S2618; intraperitoneal injection, 3 mg/kg body weight/day, administered for 1 or 4 weeks), IGF-1 (PeproTech, 250-19; intraosseous injection, 4 μg per dose per week, administered for 2 weeks), IGF-1 neutralizing antibody (R&D Systems, AF-791; intraosseous injection, 2 μg per dose, twice per week, administered for 2 or 4 weeks), VEGF neutralizing antibody (R&D Systems, AF-493-NA; intraosseous injection, 2 μg per dose, twice per week, administered for 4 weeks), PDGF-AA neutralizing antibody (R&D Systems, AF-221-NA; intraosseous injection, 2 μg per dose, twice per week, administered for 4 weeks), PDGF-BB neutralizing antibody (R&D Systems, AF-220-NA; intraosseous injection, 2 μg per dose, twice per week, administered for 4 weeks), TGF-β1 neutralizing antibody (R&D Systems, MAB2401; intraosseous injection, 2 μg per dose, twice per week, administered for 4 weeks), TGF-β2 neutralizing antibody (R&D Systems, AB-112-NA; intraosseous injection, 2 μg per dose, twice per week, administered for 4 weeks).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Derivative Assay, Flow Cytometry, Staining, Ex Vivo, In Vitro, Labeling, Expressing, Two Tailed Test

Journal: Molecular Medicine Reports

Article Title: Role and mechanism of tetrahedral DNA nanostructures in the repair of urethral injury in rats

doi: 10.3892/mmr.2026.13815

Figure Lengend Snippet: (A) Urethral tissue collection images from the control, model, model + rapamycin, and model + TDN group. (B) Body weight monitoring curves for rats in all groups over time. (C) Urethrographic imaging results for each group, with red arrows indicating the location of urethral injury model establishment. (D) Hematoxylin and eosin staining results for urethral tissues from each group. Red arrows indicate urethral lumen narrowing and associated fibrotic changes in the injured urethra. Scale bar, 500 µm (left) and 50 µm (right). TDN, tetrahedral DNA nanostructure.

Article Snippet: The rats were randomly divided into four groups: Control (n=6; intraperitoneal injection of an equal volume of saline every other day; injection volume, 0.2 ml per rat), model (n=6; urethral injury followed by intraperitoneal injection of saline every other day; injection volume, 0.2 ml per rat), model + rapamycin [n=6; 2.0 mg/kg of rapamycin (cat. no. HY-10219; MedChemExpress) injected intraperitoneally every other day after injury] and model + TDN (n=6; 10 nmol/day TDN administered via tail vein injection daily after injury; injection volume, 0.2 ml per rat).

Techniques: Control, Imaging, Staining

Journal: Molecular Medicine Reports

Article Title: Role and mechanism of tetrahedral DNA nanostructures in the repair of urethral injury in rats

doi: 10.3892/mmr.2026.13815

Figure Lengend Snippet: (A) PCA results. Different colors represent different treatment groups. (B) Sample correlation heatmap. The color intensity corresponds to correlation values. (C) Combined volcano plot showing the distribution of FCs in differentially expressed genes in the three group comparisons (model vs. control; model + rapamycin vs. model; model + TDN vs. model), with yellow dots representing upregulated genes and green dots representing downregulated genes. (D) Bar chart of differential gene counts showing the number of differential genes in the three group comparisons. (E) Venn diagram of differential genes displaying the distribution of differential genes in the three group comparisons. The numbers in different areas represent specific intersections or unique differential genes. (F) Heatmap showing the expression patterns of 25 common differentially expressed genes identified from three pairwise comparisons, displayed across four experimental groups (Control, Model, Model + rapamycin, and Model + TDN). Each row represents one gene and each column represents an individual sample. Color gradients indicate normalized gene expression levels. (G) KEGG pathway enrichment analysis of differentially expressed genes from the three pairwise comparisons (model vs. control; model + rapamycin vs. model; model + TDN vs. model). Enrichment results are presented as dot plots. The x-axis represents the GeneRatio, and the size of each dot reflects the proportion of genes enriched in the corresponding pathway. Dot color indicates the statistical significance expressed as -log10(P-value). KEGG pathways are displayed consistently across the three comparisons to facilitate direct visual comparison. KEGG, Kyoto Encyclopedia of Genes and Genomes; TDN, tetrahedral DNA nanostructure; FC, fold change; PCA, principal component analysis.

Article Snippet: The rats were randomly divided into four groups: Control (n=6; intraperitoneal injection of an equal volume of saline every other day; injection volume, 0.2 ml per rat), model (n=6; urethral injury followed by intraperitoneal injection of saline every other day; injection volume, 0.2 ml per rat), model + rapamycin [n=6; 2.0 mg/kg of rapamycin (cat. no. HY-10219; MedChemExpress) injected intraperitoneally every other day after injury] and model + TDN (n=6; 10 nmol/day TDN administered via tail vein injection daily after injury; injection volume, 0.2 ml per rat).

Techniques: Control, Expressing, Gene Expression, Comparison