Journal: Bioactive Materials

Article Title: Inhalable PD-L1-engineered hybrid cellular vesicles suppress excessive neutrophil activation and restore mitochondrial homeostasis to alleviate ischemia–reperfusion lung injury and pneumonia

doi: 10.1016/j.bioactmat.2026.03.024

Figure Lengend Snippet: Res-PD-L1@nmEVs Attenuate Inflammation and Oxidative Damage in Lung Epithelial Cells In Vitro . (A-B) Flow cytometric analysis and quantification (B) of DiO-labeled Res-PD-L1@nmEVs uptake by BEAS-2B cells under H/R conditions after pretreatment with different endocytic inhibitors (chlorpromazine, chloroquine, and filipin) or incubation at 4 °C. (C) mRNA expression levels of IL-6, TNF-α, and IL-1β in BEAS-2B cells with or without H/R injury following pretreatment with Res, nEVs, PD-L1@mEVs, PD-L1@nmEVs, or Res-PD-L1@nmEVs. (D-E) Representative fluorescence images (D) and quantitative analysis (E) of cell proliferation assessed by BrdU incorporation (red; nuclei stained with DAPI, blue). Scale bar: 50 μm. (F-G) Apoptosis rates detected by flow cytometry (F) and flow cytometric analysis of Annexin V-positive BEAS-2B cells under the indicated conditions (G). (H–K) Fluorescence microscopy images and quantitative analysis of intracellular nitric oxide (NO, green) (H-I) and reactive oxygen species (ROS, red) (J-K). Scale bar: 100 μm. (L) Flow cytometry analysis of intracellular ROS levels. (M − O) Levels of malondialdehyde (MDA) (M), superoxide dismutase 2 (SOD2) activity (N), and glutathione (GSH) content (O) in cells. (P-Q) Cell migration ability evaluated by wound healing assay under different treatments. ∗ vs. Control; # vs. H/R; & vs. H/R + PD-L1@nmEVs, p < 0.05.

Article Snippet: For cell proliferation assessment, cells were pre-treated with BrdU (40 μM, GC310002 , Servicebio) for 4 h and incubated with anti-BrdU antibody at 4 °C overnight.

Techniques: In Vitro, Labeling, Incubation, Expressing, Fluorescence, BrdU Incorporation Assay, Staining, Flow Cytometry, Microscopy, Activity Assay, Migration, Wound Healing Assay, Control

Journal: Cell Reports Medicine

Article Title: Macrophage-mimetic photothermal nanotherapeutics regulate mitochondrial homeostasis and inflammatory cascades in lung ischemia-reperfusion injury

doi: 10.1016/j.xcrm.2026.102768

Figure Lengend Snippet: Targeted repair of hypoxia/reoxygenation (H/R)-injured lung epithelial cells by Rg3@PACVs (A) Cellular uptake of Dio-labeled Rg3@PACVs (green) by H/R-injured BEAS-2B cells. Fluorescent localization was analyzed with or without pretreatment of endocytosis inhibitors (filipin, chlorpromazine, or chloroquine) (scale bars, 20 μm). (B–E) Cytokine adsorption capacity assessed by co-incubation of Rg3@PACVs (500 or 250 μg/mL) with inflammatory cytokines (IL-6, TNF-α, IL-1β, and CCL-2). Cytokine concentrations in supernatants were measured after centrifugation ( ∗∗∗ p < 0.001 vs. 0 μg/mL; # p < 0.05, ## p < 0.01 vs. 250 μg/mL; n = 3 biological replicates). (F) Cell viability of H/R-treated BEAS-2B cells assessed by cell counting kit-8 (CCK-8) assay. Cells were treated with Rg3, PACVs, or Rg3@PACVs with or without near-infrared (NIR) irradiation. (G–I) Real-time quantitative polymerase chain reaction (RT-qPCR) analysis of inflammatory gene expression (IL-6, IL-1β, and TNF-α) in H/R-stimulated BEAS-2B cells under different pretreatment conditions. (J) Immunofluorescence showing expression and co-localization of NLRP3 (green) and COX2 (red) in H/R-injured lung epithelial cells. Nuclei stained with DAPI (blue). Scale bars, 100 μm. (K) Cell proliferation detected by bromodeoxyuridine (BrdU) staining (red) in the above epithelial cells. Nuclei stained with DAPI (blue). Scale bars, 50 μm. (L) Apoptosis ratio analyzed by flow cytometry using Annexin V-Allophycocyanin (Annexin V-APC) and 7-Aminoactinomycin D (7-AAD) staining. Data are presented as the mean ± SD and analyzed using one-way ANOVA (B–E and G–I) and two-way ANOVA (F) with Tukey’s post hoc test (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 3 biological replicates, for F–I).

Article Snippet: Cell proliferation was assessed using BrdU kits ( GC310002 , Servicebio) and Cell Counting Kit-8 (CCK-8) assays (C0041, Beyotime) per manufacturer protocols.

Techniques: Labeling, Adsorption, Incubation, Centrifugation, Cell Counting, CCK-8 Assay, Irradiation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Gene Expression, Immunofluorescence, Expressing, Staining, BrdU Staining, Flow Cytometry, Control

Journal: Cell Reports Medicine

Article Title: Macrophage-mimetic photothermal nanotherapeutics regulate mitochondrial homeostasis and inflammatory cascades in lung ischemia-reperfusion injury

doi: 10.1016/j.xcrm.2026.102768

Figure Lengend Snippet: Targeted repair of hypoxia/reoxygenation (H/R)-injured lung epithelial cells by Rg3@PACVs (A) Cellular uptake of Dio-labeled Rg3@PACVs (green) by H/R-injured BEAS-2B cells. Fluorescent localization was analyzed with or without pretreatment of endocytosis inhibitors (filipin, chlorpromazine, or chloroquine) (scale bars, 20 μm). (B–E) Cytokine adsorption capacity assessed by co-incubation of Rg3@PACVs (500 or 250 μg/mL) with inflammatory cytokines (IL-6, TNF-α, IL-1β, and CCL-2). Cytokine concentrations in supernatants were measured after centrifugation ( ∗∗∗ p < 0.001 vs. 0 μg/mL; # p < 0.05, ## p < 0.01 vs. 250 μg/mL; n = 3 biological replicates). (F) Cell viability of H/R-treated BEAS-2B cells assessed by cell counting kit-8 (CCK-8) assay. Cells were treated with Rg3, PACVs, or Rg3@PACVs with or without near-infrared (NIR) irradiation. (G–I) Real-time quantitative polymerase chain reaction (RT-qPCR) analysis of inflammatory gene expression (IL-6, IL-1β, and TNF-α) in H/R-stimulated BEAS-2B cells under different pretreatment conditions. (J) Immunofluorescence showing expression and co-localization of NLRP3 (green) and COX2 (red) in H/R-injured lung epithelial cells. Nuclei stained with DAPI (blue). Scale bars, 100 μm. (K) Cell proliferation detected by bromodeoxyuridine (BrdU) staining (red) in the above epithelial cells. Nuclei stained with DAPI (blue). Scale bars, 50 μm. (L) Apoptosis ratio analyzed by flow cytometry using Annexin V-Allophycocyanin (Annexin V-APC) and 7-Aminoactinomycin D (7-AAD) staining. Data are presented as the mean ± SD and analyzed using one-way ANOVA (B–E and G–I) and two-way ANOVA (F) with Tukey’s post hoc test (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 3 biological replicates, for F–I).

Article Snippet: BrdU ELISA kit , Servicebio , Cat# GC310002.

Techniques: Labeling, Adsorption, Incubation, Centrifugation, Cell Counting, CCK-8 Assay, Irradiation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Gene Expression, Immunofluorescence, Expressing, Staining, BrdU Staining, Flow Cytometry, Control