Engineering Multifunctional Cell Membrane-Derived Nanoparticles for Hepatic Steatosis and Hepatocellular Carcinoma Therapy

Abstract

A significant challenge in pharmacotherapy is poor drug delivery, highlighting the need for platforms that efficiently deliver therapeutic agents to diseased tissues and improve efficacy. Biologically derived, engineered cell membrane-derived nanoparticles (CMNs) have emerged as promising drug-delivery systems in biomedical applications. Therefore, this thesis investigates the potential of CMNs as a drug-delivery platform to improve therapeutic efficacy for liver diseases, particularly metabolic dysfunction-associated steatotic liver disease (MASLD) and hepatocellular carcinoma (HCC). CMNs offer biocompatibility and tunable properties, with a favorable nanobiointerface that mimics native tissues, reduces immune recognition, and evades rapid clearance. Compared to synthetic nanoparticles or exosomes, CMNs are easier to produce, yield higher quantities, are noncytotoxic, and can be readily surface modified. In this thesis, CMNs were successfully fabricated via cell extrusion, with uniform shape and size confirmed by transmission electron microscopy (TEM) and other physicochemical characterization techniques. To assess CMN integrity via phosphatidylserine (PS) externalization (an ‘eat-me’ signal for phagocytic clearance), flow cytometry was performed. CMNs revealed only 1.13% PS in the outer leaflet compared with controls, indicating healthy “ghost” nanoparticles capable of prolonged systemic circulation. The CMNs demonstrated excellent drug-loading versatility and efficient cellular internalization capabilities, resulting in enhanced therapeutic efficacy. This was confirmed by loading a model drug (Rosuvastatin, RS), in which the RS-loaded CMN reduced hepatic steatosis in an in vitro hepatocyte steatosis model. Next, to achieve the targeted delivery of CMNs for hepatocyte-specific targeting, CMNs were prepared from red blood cells (RBCs) and functionalized with hepatocyte-specific targeting molecules to improve cellular specificity. The optimized CMNs were then loaded with the FDA-approved drug Resmetirom for MASLD therapy, significantly reducing hepatic steatosis in vitro. To further evaluate CMN's therapeutic potential, RBC-derived CMNs were applied to HCC treatment by incorporating the sonosensitizer indocyanine green (ICG) as a stimulus-responsive platform. In vitro evaluations showed that, upon ultrasound exposure, CMN-ICG produced 2.62-fold more reactive oxygen species (ROS) and induced mild hyperthermia (43-46 °C), ultimately leading to cancer cell death. Overall, these findings demonstrate the potential of CMNs as effective drug-delivery platforms and establish a foundation for future in vivo studies, supporting their clinical translation for liver diseases.