In Vitro Cell Work of Two Types of Targeted Nanoparticles with Ultrasound Triggering

Abstract

Targeted liposomes have shown promising potential as effective chemotherapeutics delivery vehicles in Smart Drug Delivery Systems (SDDSs). Upon accumulation and internalization at the tumor site, the liposomes need to be potentiated by an external trigger to effectively and controllably release their contents. In this study, synthesis, characterization and in vitro cell work of two types of targeted liposomes with ultrasound (US) triggering were considered. The MTT assays of PEGylated liposomes, encapsulating Doxorubicin (DOX), conjugated to human serum albumin (HSA), and Herceptin (HER) were carried out on different cell lines. The size of these nanocarriers was measured using dynamic light scattering (DLS). All three nanoparticles were found to be large unilamellar vesicles (LUVs), with radii of 83.5 ± 0.734 nm, 101 ± 1.56 nm, and 103 ± 1.86 nm for the control, HSA-conjugated, and HER-conjugated liposomes, respectively. The lipid content of the different liposomes was determined using the spectrophotometrical Stewart assay, and the confirmation of the moiety-conjugation was established using the bicinchoninic acid (BCA) assay. The MTT results revealed that functionalizing the liposomes with HSA coupled with US exposure for 20 seconds in a 40-kHz sonicating bath significantly enhanced the nanocarrier’s cellular uptake by the MCF-7 (HSA+ breast cancer) cells compared to HeLa (HSA- cervical cancer) cells, with cell viabilities of 15.7 ± 0.613 % and 47.3 ± 0.621% (p-value=8.05x10⁻⁷), respectively. As for the liposomes functionalized with HER under the same experimental conditions, the cell viabilities in SKBR-3 (HER+ breast cancer) cells and MDA-MB-231 (triple-negative breast cancer) were 27.4 ± 0.260 % and 40.1 ± 0.216% (p-value= 7.53x10⁻⁷), respectively. Thus, it is suggested that coupling US with active targeting elicits synergistic effects and enhanced drug uptake by the cells. Also, liposomal treatments, i.e., control and targeted, along with US exposure, showed more pronounced effects in both cell lines, as the observed cell viabilities were significantly less than in control non-sonicated cells. The results presented in this thesis show promise of utilizing targeted liposomal delivery and ultrasound in the treatment of cancer.