Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: Synthesis and Characterization of ROS-Responsive Hydrogel D-EVs@Gel. (A) Schematic illustration of ROS-responsive smart hydrogel fabrication process (B) Photographs demonstrating the thermosensitive sol-gel transition of D-EVs@Gel ROS formulation. (C) Degradation curve of blank Gel and D-EVs@Gel ROS under collagenase. (D) Confocal microscopy illustrating the spatial distribution of PHK26-labeled EVs (red) within D-EVs@Gel ROS . (E) Representative SEM images of Gel and D-EVs@Gel ROS . (F) Particle Size Distribution Chart for Gel and D-EVs@Gel ROS . (G) Solubility characteristics of Gel and D-EVs@Gel ROS in PBS during a duration of 48 h. (H) Rheological measurements for Gel and D-EVs@Gel ROS hydrogel under alternating high (20 %) and low (0.1 %) shear. (I) Compression test of Gel and D-EVs@Gel ROS . (J) Cumulative release profile of EVs from D-EVs@Gel incubated with PBS containing different H 2 O 2 concentrations or not. (K) Cumulative release profile of EVs from D-EVs@Gel ROS incubated with PBS containing MMP3/MMP13 or not. (L) Live/dead cytofluorograms, and (M) quantitative assessment of the cytotoxicity of Gel and D-EVs@Gel ROS .

Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibodies against GAPDH (1:5000, 104941-AP, Proteintech), TSG101 (1:1000, DF8427, Affinity), CD9 (1:1000, AF5139, Affinity), CD63 (1:2000, 25682-1-AP, Proteintech), Calnexin (1:5000, 10427-2-AP, Proteintech), GM130 (1:20000, 11308-1-AP, Proteintech), CXCR3 (1:5000, 26756-1-AP, Proteintech), CXCL10 (1:2000, 10937-1-AP, Proteintech), MMP3 (1:2000, 17873-1-AP, Proteintech), ADAMTS5 (DF13268, Affinity), P16 (AF5484, Affinity), P21 (10355-1-AP, Proteintech), GPX4 (1:1000, 381958, Zen-bio), SLC7A11 (1:1000, 26864-1-AP, Proteintech), ACSL4 (1:5000, 22401-1-AP, Proteintech) and Tubulin (1:10000, T40103 , Abmart) overnight at 4 °C.

Techniques: Formulation, Confocal Microscopy, Labeling, Solubility, Shear, Incubation

Journal: Non-coding RNA Research

Article Title: Hsa_circ_0101645 contributes to excessive autophagy and apoptosis in intervertebral disc degeneration by acting as a miR-1304-5p sponge modulating BNIP3 expression

doi: 10.1016/j.ncrna.2025.11.007

Figure Lengend Snippet: Hsa_circ_0101645 accelerating the IVDD process in vivo . A: Diagram of the animal procedure for this study. B: Grouping information for this section. C: Representative X-rays of each group of rats. Statistical graph demonstrating the disc height index (DHI) changes for L4/5 in each group of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). D: HE staining exhibiting pathological changes of CEP, NP, and AP in IVD in each group of rats (N = 6). Scale bar: 500 μm. E: EdU staining was used to detect cell proliferation in IVD tissues of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 50 μm. F: TUNEL (white light) staining exhibiting TUNEL-positive cells in IVD tissues of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 50 μm. G: The effect of hsa_circ_0101645 on the protein levels of Collagen Ⅱ, Aggrecan, MMP-3 and MMP-13 in IVD was observed by IHC staining (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 100 μm. H-I: The expression of hsa_circ_0101645 (H) and miR-1304-5p (I) in each group of IVD tissues (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). J-K: Changes in expression of apoptosis ( J; Caspase 3, Bcl-2 and Bax) and autophagy markers ( K; LC3B, Beclin and P62) in IVD tissues (N = 3) (One-way ANOVA test with Tukey's multiple comparisons test or Kruskal-Wallis test with Dunn's multiple comparisons test). ∗ indicates P < 0.05.

Article Snippet: Sections were then incubated with primary antibodies against Collagen II (28459-1-AP; 1:200; Proteintech, USA), Aggrecan (13880-1-AP; 1:100; Proteintech), MMP-3 (17873-1-AP; 1:200; Proteintech), and MMP-13 (18165-1-AP; 1:100; Proteintech), followed by the corresponding secondary antibodies.

Techniques: In Vivo, Staining, TUNEL Assay, Immunohistochemistry, Expressing

Journal: Pharmaceuticals

Article Title: Biluo Qianyuan Formula Ameliorates Post-Traumatic Osteoarthritis by Suppressing FN1-Mediated Synovial Inflammation and Restoring Joint Homeostasis

doi: 10.3390/ph19030500

Figure Lengend Snippet: Integrated network pharmacology and molecular docking analysis of candidate compounds and targets. ( A ) A Venn diagram illustrates the intersection of differentially expressed genes from the GSE178557 dataset with disease-related genes from GeneCards and drug-target genes from DrugBank. ( B ) The PPI network of overlapping targets was generated to visualize functional associations. Central nodes (red) indicate core hub genes identified based on degree centrality and connectivity. ( C ) A Sankey diagram correlates specific target genes ( left ) with their respective enriched KEGG biological pathways ( right ). Connectivity indicates involvement in signaling cascades such as the AGE-RAGE, TGF-beta, and IL-17 signaling pathways. ( D ) Representative 3D docking poses demonstrate the binding orientations and intermolecular interactions between active compounds (bavachinin, hederagenin, and myricanone) and primary protein targets (FN1, MMP3, and TGF-β). ( E ) A heatmap displays the molecular docking scores (binding energy, kcal/mol) for the interaction between candidate compounds and hub targets. Color intensity and numerical values represent the predicted binding stability.

Article Snippet: The serum concentrations of MMP3 (Cat# E-EL-M0626, Elabscience Biotechnology Co., Ltd., Wuhan, China), TGF-β (Cat# E-EL-M0051, Elabscience Biotechnology Co., Ltd., Wuhan, China), and FN1 (Cat# KE00039, Elabscience Biotechnology Co., Ltd., Wuhan, China) were quantified by ELISA using commercial kits according to the manufacturers’ instructions.

Techniques: Generated, Functional Assay, Protein-Protein interactions, Binding Assay

Journal: Pharmaceuticals

Article Title: Biluo Qianyuan Formula Ameliorates Post-Traumatic Osteoarthritis by Suppressing FN1-Mediated Synovial Inflammation and Restoring Joint Homeostasis

doi: 10.3390/ph19030500

Figure Lengend Snippet: BLQYF attenuates systemic inflammation and FN1-associated responses in PTOA. ( A ) Serum levels of MMP3, TGF-β, and FN1 in sham, PTOA, and PTOA mice treated with BLQYF or celecoxib. ( B ) Representative morphology and vimentin immunofluorescence staining of primary fibroblast-like synoviocytes (FLSs) isolated from synovial tissues of patients with post-traumatic osteoarthritis. Nuclei were counterstained with DAPI. Scale bar = 20 μm. ( C ) qRT–PCR analysis of MMP3 , TGF-β , and FN1 mRNA expression in FLSs from PTOA patients and mesenchymal stem cells (MSCs) from non-osteoarthritic hip arthroplasty donors. ( D ) CCK-8 assay showing MSC viability following BLQYF treatment (0–80 ng/mL, 24 h). ( E ) qRT–PCR analysis of MMP3 , TGF-β , and FN1 expression in FLSs following FN1 knockdown and BLQYF treatment (5, 10, 20 ng/mL, 24 h). Data were presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests. p < 0.05, p < 0.01, p < 0.001.

Article Snippet: The serum concentrations of MMP3 (Cat# E-EL-M0626, Elabscience Biotechnology Co., Ltd., Wuhan, China), TGF-β (Cat# E-EL-M0051, Elabscience Biotechnology Co., Ltd., Wuhan, China), and FN1 (Cat# KE00039, Elabscience Biotechnology Co., Ltd., Wuhan, China) were quantified by ELISA using commercial kits according to the manufacturers’ instructions.

Techniques: Immunofluorescence, Staining, Isolation, Quantitative RT-PCR, Expressing, CCK-8 Assay, Knockdown