A biohybrid nanovesicle hijacks inflammatory chemotaxis to deliver colchicine for myocardial infarction therapy

Figures

• 

  FIGURE 1. Preparation and characterization of MM-LP@COL nanovesicles. (A) FRET analysis validating the fusion between the DiO/DiI-labeled lipid membrane and the macrophage membrane. The recorded fluorescence intensity profile confirms successful integration. (B) Hydrodynamic diameter distribution of the nanovesicles measured by dynamic light scattering (DLS). Data are presented as mean ± standard deviation (n = 3). (C) Zeta potential distribution of the synthesized nanovesicles. Data are presented as mean ± standard deviation (n = 3). (D) Representative transmission electron microscopy (TEM) image revealing the morphology of MM-LP nanovesicles. Scale bar, 100 nm. (E) Confocal laser scanning microscopy (CLSM) image of MM-LP nanoparticles. A simple physical mixture of the two components served as the control. Scale bar, 50 μm. (F) Cellular uptake analysis. CLSM images of endothelial cells (ECs) following a 4-h incubation with MM-LP@DiD NPs. The MM-LP component was labeled with DiO (green), and the COL was replaced with DiD (red). Cells were pre-stimulated with TNF-α to induce expression of von Willebrand factor (vWF, aqua). Nuclei were counterstained with DAPI (blue). The composite image is…

• 

  FIGURE 2. Anti-inflammatory efficacy of MM-LP@COL nanovesicles. (A) In vitro cytokine neutralization by MM-LP nanovesicles. Neutralization capacity against TNF-α, IL-1β, IL-6, and IFN-γ was assessed following incubation with increasing concentrations of MM-LP nanovesicles (0, 1, and 3 mg/mL). Data are presented as mean ± SD (n = 3). (B) Systemic anti-inflammatory effects in a murine myocardial infarction model. Serum levels of TNF-α, IL-1β, IL-6, and CCL2 were measured by ELISA after treatment with PBS, Free COL, LP@COL, or MM-LP@COL nanovesicles (n = 6 per group). *p < 0.05, and **p < 0.01, data are presented as the mean ± SD and analyzed by using one-way ANOVA, Tukey’s multiple comparisons test.

• 

  FIGURE 3. MM-LP@COL nanovesicles reduce neutrophil levels in the blood of post-myocardial infarction mice. (A) UMAP analysis of immune cells in peripheral blood. Representative UMAP plots are shown for myocardial infarction (MI) model mice treated with PBS, Free COL, LP@COL, or MM-LP@COL nanovesicles (n = 3 per group). (B) Heatmap depicting the relative quantification of distinct immune cell populations across the different treatment groups. (C) UMAP plots illustrating the expression patterns of key immune cell markers within each identified metacluster.

• 

  FIGURE 4. MM-LP@COL nanovesicles attenuate neutrophil mobilization from the bone marrow following myocardial infarction. (A) UMAP analysis of bone marrow immune cells from MI mice treated with PBS, Free COL, LP@COL, or MM-LP@COL nanovesicles (n = 3 per group). (B) Heatmap showing the relative abundance of immune cell populations in bone marrow across treatment groups. (C) UMAP plots displaying expression patterns of characteristic surface markers within each metacluster.

• 

  FIGURE 5. MM-LP@COL nanovesicles mitigate neutrophil accumulation in the spleen after myocardial infarction. (A) UMAP analysis of splenic immune cells. Representative UMAP plots are shown for MI mice treated with PBS, Free COL, LP@COL, or MM-LP@COL nanovesicles (n = 3 per group). (B) Heatmap displaying the relative proportions of various immune cell types in the spleen across different treatment groups. (C) UMAP plots demonstrating the expression patterns of specific immune cell markers within each metacluster.

• 

  FIGURE 6. MM-LP@COL nanovesicles confer long-term therapeutic benefits after myocardial infarction. (A) Representative echocardiographic images of myocardial infarction (MI) mice treated with PBS, Free COL, LP@COL, or MM-LP@COL. (B) Quantitative analysis of cardiac function parameters: left ventricular ejection fraction (EF, left) and fractional shortening (FS, right). Data are presented as mean ± SD (n = 6). (C) Quantification of inflammatory cell infiltration in cardiac tissue based on H&E-stained sections across treatment groups. Data are presented as mean ± SD (n = 6). (D) Measurement of cardiomyocyte cross-sectional area from WGA-stained sections. Data are presented as mean ± SD (n = 6). (E) Representative photomicrographs of H&E-stained myocardial tissue. Scale bar, 500 μm. (F) Representative fluorescent images of WGA-stained myocardial tissue, outlining cardiomyocyte membranes. Scale bar, 20 μm. *p < 0.05, **p < 0.01, and ***p < 0.001, data are presented as the mean ± SD and analyzed by using one-way ANOVA, Tukey’s multiple comparisons test.

• 

  FIGURE 7. MM-LPs nanovesicles demonstrate favorable biosafety. (A–D) Biochemical markers relevant to hepatic and kidney function. Data are presented as mean ± SD (n = 6 per group). (E) Hematoxylin and eosin (H&E) staining of key organs from mice treated with LP nanoparticles and MM-LP nanoparticles, scale bar, 100 μm.