Preparation of M2 macrophage membrane hybrid lipid nanoparticles based on microfluidic device for atherosclerosis treatment.

With the continuous advancement of bionanomaterial technology, the design and fabrication strategies of drug delivery systems have undergone significant strategic transformations and innovations. Herein, we report a microfluidic chip that enables one-step production of cell membrane hybrid lipid nanoparticles by fusing synthetic phospholipids with M2-macrophage membrane fragments and simultaneously encapsulating simvastatin and rapamycin. This greatly simplifies the preparation process, improves efficiency and produces M2-macrophage-membrane hybrid lipid nanoparticles loaded with simvastatin and rapamycin (M2LNPs@SIM&RAPA) with small particle size, homogeneous distribution and enhanced cumulative release in vitro. Western blotting and SDS-PAGE confirm the successful embedding of M2-membrane proteins. The formulation merges the long-circulating stability of synthetic phospholipids with the inflammation-targeting capacity and biocompatibility of M2 macrophage membranes. Stability tests and cell uptake experiments both proved its excellent storage stability and inflammation targeting. Cell cytotoxicity tests optimized the safe dosages of simvastatin and rapamycin. In the subsequent experiments, M2LNPs@SIM&RAPA could significantly reduce the expression of TNF-α in inflammatory cells and inhibit the formation of lipid droplets in foam cells, demonstrating its excellent anti-inflammatory activity and enhanced cholesterol excretion ability. In summary, the M2LNPs@SIM&RAPA prepared based on microfluidics can precisely regulate lipid metabolism and inflammatory responses through the synergistic effect of low-dose drugs, and is expected to provide a theoretical basis for the future treatment of atherosclerosis.