Improvement of cell membrane permeability using a cell-solution electrode for generating atmospheric-pressure plasma

This study reports the potential of cold atmospheric plasma (CAP) as a versatile tool for delivering oligonucleotides into mammalian cells. Compared to lipofection and electroporation methods, plasma transfection showed a better uptake efficiency and less cell death in the transfection of oligonucleotides. We demonstrated that the level of extracellular aqueous reactive oxygen species (ROS) produced by gas plasma is correlated with the uptake efficiency and that this is achieved through an increase of intracellular ROS levels and the resulting increase in cell membrane permeability. This finding was supported by the use of ROS scavengers, which reduced CAP-based uptake efficiency. In addition, we found that cold atmospheric plasma could transfer oligonucleotides such as siRNA and miRNA into cells even in 3D cultures, thus suggesting the potential for unique applications of CAP beyond those provided by standard transfection techniques. Together, our results suggest that cold plasma might provide an efficient technique for the delivery of siRNA and miRNA in 2D and 3D culture models.

Section: Methodsmentioning

confidence: 99%

“…However, Sakai reported that the plasma could induce DNA strand breaks in a PBS solution27. A recent study showed that by using the cell-solution electrode, plasma jet could improve cell permeability compared with the conventional diffusion type plasma31. Some publications have discussed transdermal drug delivery by cold atmosphere plasma323334.…”

mentioning

confidence: 99%

Intracellular ROS mediates gas plasma-facilitated cellular transfection in 2D and 3D cultures

Wang

et al. 2016

“…observed the improvement of the cell membrane permeability using a cell-solution electrode CAP and they attributed this to the increase of the electric field and the density of the short-lived ROS reaching the cells in the solution 11 . Moreover, Hong et al .…”

Section: Introductionmentioning

confidence: 99%

Effect of head group and lipid tail oxidation in the cell membrane revealed through integrated simulations and experiments

Yusupov

Wende

Kupsch

et al. 2017

We report on multi-level atomistic simulations for the interaction of reactive oxygen species (ROS) with the head groups of the phospholipid bilayer, and the subsequent effect of head group and lipid tail oxidation on the structural and dynamic properties of the cell membrane. Our simulations are validated by experiments using a cold atmospheric plasma as external ROS source. We found that plasma treatment leads to a slight initial rise in membrane rigidity, followed by a strong and persistent increase in fluidity, indicating a drop in lipid order. The latter is also revealed by our simulations. This study is important for cancer treatment by therapies producing (extracellular) ROS, such as plasma treatment. These ROS will interact with the cell membrane, first oxidizing the head groups, followed by the lipid tails. A drop in lipid order might allow them to penetrate into the cell interior (e.g., through pores created due to oxidation of the lipid tails) and cause intracellular oxidative damage, eventually leading to cell death. This work in general elucidates the underlying mechanisms of ROS interaction with the cell membrane at the atomic level.In recent years, cold atmospheric plasmas (CAPs) are gaining increasing interest for cancer treatment, i.e., so-called "plasma oncology" [1][2][3][4] . In a recent review, Schlegel et al. 1 summarize the results of several studies on plasma oncology and show the progress in the potential use of CAPs to effectively kill cancer cells (either by apoptosis or necrosis), in vitro as well as in vivo. CAP sources seem to be a powerful tool for cancer treatment, either alone or in combination with other conventional therapies. Indeed, recent experimental results showed that CAPs could enhance the effects of conventional chemotherapy even in resistant tumorous cells; the resistant cell population, if pre-treated with CAP, becomes sensitive to treatment with chemotherapy 5,6 . Moreover, it was demonstrated that plasma treatment, both in vitro and in vivo, is able to attack a wide range of cancer cell lines without damaging their normal counterparts 1, 2 . Thus, preliminary results seem very promising. Nevertheless, the application of CAPs for cancer treatment is still in its initial stage, and there is an enormous need for a better understanding of the underlying mechanisms.CAPs generate reactive oxygen species (ROS, e.g., OH, HO 2 , H 2 O 2 ) and reactive nitrogen species (RNS, e.g., NO, NO 2 , ONOO − ), which are generally believed to play a key role in plasma treatment 3,4,7 . Several studies showed that CAPs elevate intracellular ROS levels, thereby inducing oxidative damage in cancer cells, which can lead to cell death, i.e., apoptosis 3,8 . Normal cells, on the other hand, are able to defend themselves from this harmful effect of ROS by activating multiple anti-oxidative systems that reduce the increased oxidative stress and restore the balance 9, 10 . Besides, also the RNS generated by CAP might play an important role in cancer therapy (see ref. 4 and references therein). ...

“…Plasma medicine is a rapidly emerging field, and a number of researchers have reported innovative applications of non-equilibrium atmospheric-pressure plasma (APP) 1 2 3 4 , including its use in the selective killing of cancer cells 5 , in blood coagulation for minimally invasive surgery, in assisting wound healing and tissue regeneration 6 , and as a gene transfer tool 7 8 9 . Non-equilibrium APP has a higher electron temperature (~several eV) than gas (ion) temperature 10 .…”

mentioning

confidence: 99%

Calcium influx through TRP channels induced by short-lived reactive species in plasma-irradiated solution

Sasaki

Kanzaki

Kaneko

2016

Self Cite

Non-equilibrium helium atmospheric-pressure plasma (He-APP), which allows for a strong non-equilibrium chemical reaction of O2 and N2 in ambient air, uniquely produces multiple extremely reactive products, such as reactive oxygen species (ROS), in plasma-irradiated solution. We herein show that relatively short-lived unclassified reactive species (i.e., deactivated within approximately 10 min) generated by the He-APP irradiation can trigger physiologically relevant Ca2+ influx through ruthenium red- and SKF 96365-sensitive Ca2+-permeable channel(s), possibly transient receptor potential channel family member(s). Our results provide novel insight into understanding of the interactions between cells and plasmas and the mechanism by which cells detect plasma-induced chemically reactive species, in addition to facilitating development of plasma applications in medicine.