INVESTIGATION OF CELL MEMBRANE PERMEABILIZATION INDUCED BY PLASMA TREATMENT FOR DRUG DELIVERY APPLICATIONS
Résumé
Cold atmospheric pressure plasmas have demonstrated their ability in biomedical applications thanks to their low gas temperature and their capacity to produce radicals, ions, electrons, UV radiation and electric field. However, the understanding of the interactions between plasma, living cells and tissues is still far from being completely understood. Several studies have shown that plasma could be used to enhance cell permeabilization to deliver chemotherapy [1] or nucleic acids [2].
In this work, we investigated the parameters needed for helium plasma to induce a cell membrane permeabilization on cancerous cells for drug delivery purposes. Using our optimal settings, different fluorescent molecular markers were used in order to determine pore size, opening kinetics and the importance of endocytosis in the process. We were able to deliver propidium iodide in almost 30% of HeLa cells 30 minutes after plasma treatment generated at 14kV, a frequency of 100Hz during 100s using 0.5slm of helium. No significant cytotoxicity was measured 24h after treatment. Efficient drug delivery was obtained using molecules up to 70kDa (figure 1). Experiments using plasma activated media were done to understand the contribution of radical formation on cell membrane permeabilization. The influence of plasma-generated electric field has also been investigated. For this purpose, a dielectric barrier was used between the plasma and the biological target. A comparison with and without the barrier will be presented for biological application. In conclusion drug delivery using plasma is dependent of the electric field, endocytosis and plasma-induced chemistry.
Figure 1. Fluorescence microscopy of HeLa cells 30 minutes after treatment by helium plasma in the presence of propidium iodide, 70kDa FITC-Dextran or 150kDa FITC-Dextran.
References
[1] W. Zhu, et al., Scientific Report, 6, 21974 (2016).
[2] M. Jinno, et al., Arch. Biochem. Biophys, 605, 59 (2016).