Combination effect of cold atmospheric plasma with green synthesized zero-valent iron nanoparticles in the treatment of melanoma cancer model.

Green synthesized zero-valent iron nanoparticles (nZVI) have high potential in cancer therapy. Cold atmospheric plasma (CAP) is also an emerging biomedical technique that has great potential to cure cancer. Therefore, the combined effect of CAP and nZVI might be promising in treatment of cancer. In this study, we evaluated the combined effect of CAP and nZVI on the metabolic activity of the surviving cells and induction of apoptosis in malignant melanoma in comparison with normal cells. Therefore, the effect of various time exposure of CAP radiation, different doses of nZVI, and the combined effect of CAP and nZVI were evaluated on the viability of malignant melanoma cells (B16-F10) and normal fibroblast cells (L929) at 24 h after treatment using MTT assay. Then, the effect of appropriate doses of each treatment on apoptosis was evaluated by fluorescence microscopy and flow cytometry with Annexin/PI staining. In addition, the expression of BAX, BCL2 and Caspase 3 (CASP3) was also assayed. The results showed although the combined effect of CAP and nZVI significantly showed cytotoxic effects and apoptotic activity on cancer cells, this treatment had no more effective compared to CAP or nZVI alone. In addition, evaluation of gene expression showed that combination therapy didn't improve expression of apoptotic genes in comparison with CAP or nZVI. In conclusion, combined treatment of CAP and nZVI does not seem to be able to improve the effect of monotherapy of CAP or nZVI. It may be due to the resistance of cancer cells to high ROS uptake or the accumulation of saturated ROS in cells, which prevents the intensification of apoptosis.