Targeted gold nanorods combined with low-intensity nsPEFs enhance antimelanoma efficacy in vitro

High-intensity nanosecond pulsed electric fields (nsPEFs) are a novel treatment with promising applications for cell stimulation and tissue ablation, and many research studies have shown that gold nanorods (GNRs) are high-conductivity nanomaterials that can enhance electroporation for biomedical applications. In addition, the folic acid (FA) receptor has been demonstrated as a valuable therapeutic target that is highly expressed in a variety of cancers. To reduce the electric field strength required to treat tumors by nsPEFs, for the first time, gold nanorods with folic acid were proposed to achieve higher antimelanoma efficacy at lower electric field intensity in this study. The surface of polyethylene glycol-gold nanorods with good biocompatibility was further modified by folic acid (FA) to provide modified gold nanorods (GNR-PEG-FA) with specific targeted recognition of A375 melanoma cells. The binding of GNRs to A375 melanoma cells was observed by dark field microscopy. After combined treatment with nsPEFs and GNRs, cell viability was evaluated by a CCK-8 assay. Flow cytometry was performed to evaluate apoptosis and the cell cycle. And active caspase 3 was also detected after treatment. The antimelanoma efficacy was enhanced in a pulsed electric field-dependent manner. More importantly, compared with the group of nsPEFs alone and gold nanorods without FA, treating cells with nsPEFs combined with GNR-PEG-FA resulted in a lower percentage of viable cells, higher percentages of necrosis and apoptosis and higher concentration of active caspase 3 and induced cell cycle arrest in S phase, effectively inhibiting the proliferation of A375 melanoma cells. nsPEFs combined with GNR-PEG-FA showed the best antimelanoma efficacy in vitro and effectively killed melanoma cells with low-intensity nsPEFs. The combined treatment of cells with nsPEFs and GNR-PEG-FA is expected to become a safer and more efficient physical treatment of melanomas. (paper)