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