Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: In vitro and in vivo biological effects and targeting characterization of PFNBs. (A) Schematic diagram of the endothelial barrier penetration ability evaluation of PFNBs, and (B) quantitative analysis results (n = 3). (C) Schematic diagram of the enhanced uptake effect of PFNBs by microglia with/without inflammatory activation. (D) confocal microscopy images, and (E) quantitative analysis results (n = 3). Scale bar, 10 μm. (F) Comparison of the PNBs and PFNBs uptaken by microglia with/without inflammatory activation at 4 h (n = 3). (G) Near-infrared fluorescence images of PFNBs targeting AIS lesions, and (H) quantitative analysis results (n = 3). (I) Near-infrared fluorescence images of PFNBs distribution in brains and major organs, as well as (J) quantitative analysis results (n = 3). Error bars: mean ± standard deviation. ns indicates non-significant (p > 0.05). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (two-tailed Student's t -test).

Article Snippet: Then, the fluorescence signals from the head region were acquired at predetermined time points (Pre, 0.5, 1, 2, 4, 8, 12, and 24 h) using a near-infrared fluorescence in vivo imaging system (IVIS Spectrum Imaging System, Caliper Life Sciences, USA).

Techniques: In Vitro, In Vivo, Activation Assay, Confocal Microscopy, Comparison, Fluorescence, Standard Deviation, Two Tailed Test

Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: Evaluation of the immunomodulatory effects of PFNBs in vitro and in vivo . (A) Confocal fluorescence microscopy images of iNOS and CD206 immunofluorescence staining in inflammatory activated microglia after being incubated with Saline, free FTY720, PNBs, and PFNBs. Scale bar, 50 μm. (B) Quantitative analysis of iNOS (M1) and CD206 (M2) fluorescence intensity based on confocal images (n = 3). (C) WB bands of iNOS, CD206, p-STAT3 and t-STAT3 proteins. (D) Quantification of iNOS/GAPDH, CD206/GAPDH and p-STAT3/t-STAT3 levels,/Saline represents the change of experimental group relative to Saline group (n = 3). (E) Immunofluorescence staining and quantitative analysis of M1 (F) and M2 (G) phenotypes of microglia in the ischemic lesion (n = 5). Scale bar: 50 μm. (H) WB bands of iNOS, CD206, p-STAT3, and t-STAT3 proteins in lesion brain tissues of AIS mice. (I) Quantification of iNOS/GAPDH, CD206/GAPDH and p-STAT3/t-STAT3 levels in lesion brain tissues of AIS mice. Saline represents the change of experimental group relative to Saline group (n = 3). Error bars: mean ± standard deviation. ns indicates non-significant (p > 0.05). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (two-tailed Student's t -test).

Article Snippet: Then, the fluorescence signals from the head region were acquired at predetermined time points (Pre, 0.5, 1, 2, 4, 8, 12, and 24 h) using a near-infrared fluorescence in vivo imaging system (IVIS Spectrum Imaging System, Caliper Life Sciences, USA).

Techniques: In Vitro, In Vivo, Fluorescence, Microscopy, Immunofluorescence, Staining, Incubation, Saline, Standard Deviation, Two Tailed Test

Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: Evaluation of the protective effects on BBB permeability and vascular integrity of PFNBs in vitro and in vivo . (A) Schematic time line protocol of in vitro BBB model establishment and subsequent operations. (B) TEER value monitoring of endothelial cell monolayers after OGD/R treatment with addition of Saline, free FTY720, PNBs, and PFNBs (n = 3). (C) Proteomic analysis of the species and expression abundance of growth factors carried by platelet membranes in PLTs, PMVs, PNBs, and PFNBs (n = 3). (D) TEER value monitoring of endothelial cell monolayers seeded with inflammatory activated microglia in the lower chamber after OGD/R treatment with addition of Saline, free FTY720, PNBs, and PFNBs (n = 3). Confocal fluorescence microscopy images of CD34 and ZO-1 (E) or occluding (F) double-labeled immunofluorescence staining in the lesion area 24 h after AIS modeling and administration. Scale bar, 20 μm. Quantitative analysis of the ratio of ZO-1/CD34 positive area (G) or occludin/CD34 positive area (H) (n = 5). Photos of AIS mice brains in each group after Evans blue injection (I) and quantitative analysis of Evans blue leakage (J) (n = 3). (K) Quantitative analysis of brain edema in each group of AIS mice (n = 3). Error bars: mean ± standard deviation. ns indicates non-significant (p > 0.05). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (two-tailed Student's t -test).

Article Snippet: Then, the fluorescence signals from the head region were acquired at predetermined time points (Pre, 0.5, 1, 2, 4, 8, 12, and 24 h) using a near-infrared fluorescence in vivo imaging system (IVIS Spectrum Imaging System, Caliper Life Sciences, USA).

Techniques: Permeability, In Vitro, In Vivo, Saline, Expressing, Fluorescence, Microscopy, Labeling, Immunofluorescence, Staining, Injection, Standard Deviation, Two Tailed Test