Cold atmospheric plasma mediated cell membrane permeation and gene delivery-empirical interventions and pertinence

Delivery of genetic material to cells is an integral tool to analyze and reveal the genetic interventions in normal cellular processes and differentiation, disease development and for gene therapy. It has profound applications in pharmaceutical, agricultural, environmental and biotechnological sectors. The major methods relied for gene delivery or transfection requires either viral vectors or xenogenic carrier molecules, which renders probabilistic carcinogenic, immunogenic and toxic effects. A newly evolved physical method, Cold atmospheric plasma induced transfection neither needs vector nor carriers. The 4th state of matter 'Plasma' is a quasineutral ionized gas-containing ions, neutral atoms, electrons and reactive radical molecules; and possess electric and magnetic field, along with emanating photons and UV radiations. Plasma produced at atmospheric pressure conditions, and having room temperature is conferred as Low temperature plasma or Cold atmospheric plasma. Selective and controlled application of cold atmospheric plasma on tissues creates temporary, restorable pores on cell membranes that could be diligently manipulated for gene delivery. Research in this regard attained pace since 2016. Cold atmospheric plasma induces transfection by lipid peroxidation, electroporation, and clathrin dependent endocytosis in cell membranes, by virtue of its reactive radicals and electric field. Plasma formed reactive radicals, especially 'OH' penetrates to the cell membrane and cleaves the phosphate head group of membrane lipids, peroxidize and detaches fatty acid tails. This decreases membrane thickness, increases membrane fluidity and permeability. Simultaneously plasma formed ions, electrons and reactive radicals accumulate over cells, generating local electric field and neutralize the negative charge of cell membrane. This induces stress on cell membrane and disrupts its structural integrity, by infringing the dynamic equilibrium between surface tension, spatial repulsion and linear tension between the head groups of phospholipids, generating minute pores. Neutralization of membrane charge promote foreign, external plasmid and gene movement towards cells and its enhanced binding with ligands and receptors on cell membrane, instigating clathrin dependent endocytosis. In vitro and in vivo studies have successfully delivered plasmids, linear DNA, siRNA and miRNA to several established cell lines like, HeLa, PC12, CHL, HUVEC, Jurkat, MCF, SH-SY5Y, HT, B16F10, HaCaT, LP-1, etc., and live C57BL/6 and BALB/c mice, using cold atmospheric plasma. This review delineates the cell surface mechanism of plasma-induced transfection, critically summarizes the research progress in this context, plasma devices used, and the inimitable features of this method. Metabolic activity, cell function, and viability are not adversely affected by this process; moreover, the cell permeating plasma-formed reactive radicals are effectively defended by cellular antioxidant mechanisms like superoxide dismutase, glutathione reductase and cytokines, alleviating its toxicity. A deeper understanding on mechanism of plasma action on cells, its aftermath, and the research status in this field would provide a better insight on future avenues of research.