Exosomes fused liposomes: formulation, stability studies and theranostic evaluation for breast cancer applications

This study presents exosomes fused liposomes vesicles as a multifunctional nanocarriers for breast cancer theranostics. Exosomes isolation from human plasma using polyethylene glycol (PEG) precipitation is an efficient and cost-effective method that preserves vesicle integrity. These exosomes were fused with liposomes to enhance drug loading, stability, and tumor-targeting. The PLB-EXO-LIPO-VCs and MB-EXO-ICG-LIPO-VCs fused vesicles showed particle sizes of 194.5 ± 2.8 nm and 175.0 ± 2.1 nm, PDI values of 0.234 ± 0.01 and 0.225 ± 0.02, and zeta potentials of - 30.31 ± 0.05 mV and - 28.33 ± 0.04 mV, respectively. TEM, SEM, and SPM confirmed a spherical morphology with smooth membranes, and FTIR analysis validated membrane fusion, with retention of exosomal components. The PLB-EXO-LIPO-VCs exhibited encapsulation efficiency and drug loading (EE/DL) of 86.78 ± 0.05 % and 22.01 ± 0.02 %, while EE/DL of MB-EXO-ICG-LIPO-VCs showed 94.06 %/27.45 % for ICG and 78.91 %/20.23 % for MB. The vesicles demonstrated pH-responsive, sustained drug release governed by anomalous (non-Fickian) diffusion. The fused vesicles showed superior stability at 25 °C, 4 °C, and -20 °C. Furthermore, PLB-EXO-LIPO-VCs exhibited 19.53-fold higher cytotoxicity than free PLB, accompanied by increased cellular uptake, ROS generation, and mitochondria-mediated apoptosis. Safety of the vesicles was confirmed by haematological, biochemical, haemolysis, and histopathological evaluations. In vivo imaging revealed tumor-specific accumulation and significant reduction in tumor volume, hypoxia, and angiogenesis in DMBA-induced breast tumors, highlighting their potential in breast cancer diagnostic and therapeutic applications.