A Reproducible Workflow for Macrophage Membrane Isolation and Nanocore Selection Toward Bioinspired Nanoparticles...

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Immunology

Background:

• Macrophage membrane-coated nanoparticles (M2M-NPs) are promising for treating
• cold
• tumors resistant to conventional therapies.
• Current methods for membrane isolation and NP coating lack reproducibility, hindering clinical translation.

Purpose of the Study:

• To establish a reproducible workflow for isolating M2 macrophage plasma membranes.
• To develop and characterize a reliable method for preparing M2M-NPs.
• To evaluate the efficacy of M2M-NPs loaded with paclitaxel against specific cancer types.

Main Methods:

• Optimized workflow combining hypotonic lysis, Dounce homogenization, and differential centrifugation for membrane isolation.
• Development and characterization of membrane-derived nanovesicles (M2M-NVs) using fluorescent lipids and cholesterol.
• Systematic evaluation of different nanocores (TPGS, VD3, PLGA) for M2M coating efficiency, with PLGA NPs yielding optimal results.

Main Results:

• Consistent M2M fractions with reproducible protein, lipid, and DNA profiles were obtained.
• PLGA/M2M-NPs demonstrated high colloidal stability, hemocompatibility, and immune evasion.
• Paclitaxel-loaded PLGA/M2M-NPs exhibited selective cytotoxicity against triple-negative breast cancer, pancreatic, and glioblastoma cells.

Conclusions:

• A reproducible workflow for engineering macrophage membrane-based biomimetic NPs was established.
• The developed M2M-NPs show enhanced therapeutic potential for immunologically
• cold
• tumors.
• This work advances the translational potential of biomimetic nanoparticles in cancer therapy.