Preparation and characterization of the engineered EVs (TPCS@EV). (A) Western blot analysis of the positive markers (i.e., CD9 and CD63) and negative markers (i.e., calnexin and APOB100) of M1 macrophage-derived EVs. TEM imaging (B) and nanoparticle tracking analysis (C) of EVs. (D) Fluorescence colocalization analysis of TPCS@EV by CLSM. Red, TPCS nano-complexes. Green, 3,3–dioctadecyloxacarbocyanine perchlorate (DiO)-labeled EVs. (E) Fluorescence colocalization analysis of TPCS@EV by NanoView. Red, TPCS. Green, CD81-positive EVs. Blue, CD9-positive EVs. TEM imaging (F) and dynamic light scattering analysis (G) of TPCS and TPCS@EV. Data in G are represented as mean ± SD (n = 3).