Journal: Journal of Translational Medicine

Article Title: Platelet membrane-coated nanoparticles inhibit platelet activation and neutrophil extracellular traps formation in acute lung injury

doi: 10.1186/s12967-025-06649-2

Figure Lengend Snippet: Preparation and characterization of PNPs ( A ) Schematic illustration of PNP preparation. ( B ) Transmission electron microscopy image of uranyl acetate-stained PNPs (scale bar: 100 nm). ( C ) Dynamic diameter and ( D ) zeta potential of PNPs were assessed by a Malvern particle size analyzer ( n = 3). ( E ) The hydrodynamic diameter of PNPs in mouse serum at 4 or 37 °C. ( F ) Fluorescence images of the Dil-labeled PNPs (red) coincubated with Hoechst-stained neutrophils (scale bar: 50 μm). ( G ) Western blotting analysis of CD62P levels in platelet lysates, platelet vesicles and PNPs

Article Snippet: Blood was collected in ethylenediaminetetraacetic acid (EDTA) tubes, and neutrophils were isolated with the Human Neutrophil Isolation Kit (Solarbio, China) according to the manufacturer’s instructions.

Techniques: Transmission Assay, Electron Microscopy, Staining, Zeta Potential Analyzer, Fluorescence, Labeling, Western Blot

Journal: Journal of Translational Medicine

Article Title: Platelet membrane-coated nanoparticles inhibit platelet activation and neutrophil extracellular traps formation in acute lung injury

doi: 10.1186/s12967-025-06649-2

Figure Lengend Snippet: PNPs inhibited thrombin-mediated platelet activation and reduced NETs formation. ( A, B ) Flow cytometry analysis of CD41 and CD62P expression in human platelets preincubated with PNPs or RBC-NPs followed by PBS or thrombin treatment. (C, D) Quantitative analysis showing ( C ) the percentage of CD62P-positive platelets and ( D ) the mean fluorescence intensity (MFI) of CD62P. ( E, F ) The neutrophils were cocultured with differentially treated platelets and subsequently stained with SYTOX Green, with fluorescence intensity quantified using a high-throughput multimodal system, and NETs formation was visualized by fluorescence microscopy (scale bar: 50 μm). ( G ) Immunostained neutrophils (MPO/citH3) after coculture with processed platelets from different treatment groups were visualized by fluorescence microscopy (scale bar: 100 μm). The data are presented as the means ± standard errors of the means (SEMs). ns, nonsignificant; *** p < 0.001

Article Snippet: Blood was collected in ethylenediaminetetraacetic acid (EDTA) tubes, and neutrophils were isolated with the Human Neutrophil Isolation Kit (Solarbio, China) according to the manufacturer’s instructions.

Techniques: Activation Assay, Flow Cytometry, Expressing, Fluorescence, Staining, High Throughput Screening Assay, Microscopy

Journal: Journal of Translational Medicine

Article Title: Platelet membrane-coated nanoparticles inhibit platelet activation and neutrophil extracellular traps formation in acute lung injury

doi: 10.1186/s12967-025-06649-2

Figure Lengend Snippet: PNPs reduced mitochondrial superoxide production and inhibited Syk phosphorylation in neutrophils. ( A, B ) Fluorescence microscopy (scale bar: 100 μm) and flow cytometry analysis of mitoSOX-stained neutrophils after coculture with thrombin-activated platelets pretreated with PNPs or RBC-NPs. ( C ) Western blotting analysis of neutrophil protein expression showing phosphorylated Syk (p-Syk), total Syk, and GAPDH expression levels. p-Syk and Syk band intensities were quantified using ImageJ software. ( D, E ) Flow cytometry analysis of CD62P expression in platelets treated with or without a CD62P-neutralizing antibody. ( F, G ) Platelets were mixed with PNPs or an anti-CD62P antibody, treated with thrombin and centrifuged to obtain processed platelets. Neutrophils were either treated with mitoTEMPO or left untreated before coincubation with the processed platelets. The cocultures were stained with SYTOX Green, the fluorescence intensity was quantified using a high-throughput multimodal system, and NETs formation was visualized by fluorescence microscopy (scale bar: 50 μm). ns, nonsignificant; *** p < 0.001

Article Snippet: Blood was collected in ethylenediaminetetraacetic acid (EDTA) tubes, and neutrophils were isolated with the Human Neutrophil Isolation Kit (Solarbio, China) according to the manufacturer’s instructions.

Techniques: Phospho-proteomics, Fluorescence, Microscopy, Flow Cytometry, Staining, Western Blot, Expressing, Software, High Throughput Screening Assay

Journal: Journal of Translational Medicine

Article Title: Platelet membrane-coated nanoparticles inhibit platelet activation and neutrophil extracellular traps formation in acute lung injury

doi: 10.1186/s12967-025-06649-2

Figure Lengend Snippet: PNP treatment reduced MPO levels and decreased peripheral blood neutrophil percentages in mice with ALI. (A) Experimental design for establishing the ALI model and treatment the mice with PNPs. The mice received intratracheal administration of PNPs or RBC-NPs followed by intranasal LPS challenge after 1 h, and samples were collected at 24 h. (B) Plasma MPO levels were measured by ELISA. (C) The percentages of neutrophils and CXCR2 + neutrophils in the blood were measured using flow cytometry analysis. (D) Complete blood count data, including the neutrophil absolute value count (NEUT), monocyte absolute value count (MONO), platelet count (PLT) and lymphocyte absolute value count (LYM), were recorded. The data are presented as the means ± standard errors of the means (SEMs). ns, nonsignificant, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Blood was collected in ethylenediaminetetraacetic acid (EDTA) tubes, and neutrophils were isolated with the Human Neutrophil Isolation Kit (Solarbio, China) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: Platelet membrane-coated nanoparticles inhibit platelet activation and neutrophil extracellular traps formation in acute lung injury

doi: 10.1186/s12967-025-06649-2

Figure Lengend Snippet: PNP treatment alleviated acute lung inflammation and injury. The mice were intratracheally challenged with either PNPs or RBC-NPs, followed by intranasal administration of LPS. ( A ) BALF cells were subjected to Giemsa staining and counted across 3 microscopic fields (scale bar: 100 μm). ( B ) Total protein concentration in BALF was measured using a BCA assay kit. ( C ) IL-1β, IL-6 and TNF-α levels in the BALF were measured by ELISA. ( D, E ) Representative lung tissue sections stained with H&E and Sirius Red (scale bar: 100 μm). ( F-H ) The percentages of neutrophils, CXCR2 + neutrophils and F4/80 + macrophages in the BALF were measured using flow cytometry analysis. The data are presented as the means ± standard errors of the means (SEMs). ns, nonsignificant, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Blood was collected in ethylenediaminetetraacetic acid (EDTA) tubes, and neutrophils were isolated with the Human Neutrophil Isolation Kit (Solarbio, China) according to the manufacturer’s instructions.

Techniques: Staining, Protein Concentration, BIA-KA, Enzyme-linked Immunosorbent Assay, Flow Cytometry