New insights on molecular internalization and drug delivery following plasma jet exposures

In this study, we evaluated cold atmospheric plasmas as a physical drug delivery tool for human cervical cancer HeLa cells and murine breast carcinoma 4T1 cells. Different cell exposure protocols - plasma jet, plasma treated medium, and combinations of plasma-induced electric field and plasma treated medium- have been proposed and assessed to provide new insight on plasma-induced uptake mechanism. Cell culture medium composition and volume are key parameters to achieve an efficient molecular uptake. The plasma device enabled the delivery of molecules having 150 kDa-size into 4Tlcells. For the first time to our knowledge, substance uptake kinetics after plasma treatment were investigated. The percentage of positive cells for propidium iodide and an anticancer agent, doxorubicin, was higher when the drugs were added a few minutes after treatment. The Plasma treated medium was not found to be as efficient as direct plasma treatment in 4T1 cells while allowing an efficient delivery in HeLa cells. Uptake levels as high as 39.3 +/- 2.9% and 40.1 +/- 9.5% for HeLa and 4 T1 cells respectively were achieved for optimized operating conditions, for which the viability of the cells was not severely affected. We also observed that plasma treatment induced the formation of actin stress fibers into cells revealing a mechanical stress.