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Figure 2. Characterization of PgNVs and their inhibitory effects on LPS-induced proinflammatory cytokine expression in RAW 264.7 macrophages. (A) Schematic representation of PgNVs isolation using differential centrifugation, TFF, and SEC; (B) Representative TEM obtained from PgNVs; (C) Representative NTA images obtained from PgNVs; (D) Determinations of particles/protein ratio and mode size by NTA and protein quantification; (E) Relative mRNA expression of Tnfa, Il6 and Il1b in RAW 264.7 cells incubated with LPS or LPS and PgNVs (n = 3). Tbp was used as a housekeeping gene. Measurements expressed as mean ± SD. *Significantly different from control group, P < 0.05; **significantly different from control group, P < 0.01; ^#significantly different from LPS group, P < 0.05; ^##significantly different from LPS group P < 0.01. Created in BioRender. MARCILLA, A. (2026). https://BioRender.com/pi0m2pq. EVs: Extracellular vesicles; LPS: lipopolysaccharide; RAW: murine macrophage cell line RAW 264.7; PgNVs: pomegranate-derived nanovesicles; TFF: tangential flow filtration; SEC: size exclusion chromatography; TEM: transmission electron microscopy; NTA: nanoparticle tracking analysis; mRNA: messenger RNA; Tnfa: tumor necrosis factor alpha; Il6: interleukin 6; Il1b: interleukin 1 beta; Tbp: TATA-box binding protein; SD: standard deviation.