Intracellular ROS mediates gas plasma-facilitated cellular transfection in 2D and 3D cultures.
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.
Centre for Plasma Biomedicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.|State Key Laboratory of Electrical Insulation and Power Equipment, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.
Dehui
D
Xu
Centre for Plasma Biomedicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.|State Key Laboratory of Electrical Insulation and Power Equipment, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.
Biqing
B
Wang
Centre for Plasma Biomedicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.|State Key Laboratory of Electrical Insulation and Power Equipment, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.
Yujing
Y
Xu
Centre for Plasma Biomedicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.|State Key Laboratory of Electrical Insulation and Power Equipment, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.
Zeyu
Z
Chen
Centre for Plasma Biomedicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.|State Key Laboratory of Electrical Insulation and Power Equipment, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.
Qinjie
Q
Cui
Department of Cardiovascular Medicine, First Affiliated Hospital of the Medical School, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.
Yanjie
Y
Yang
Frank Reidy Center for Bioelectrics, Old Dominion University, Norfolk, VA, 23508, USA.
Hailan
H
Chen
Centre for Plasma Biomedicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.|State Key Laboratory of Electrical Insulation and Power Equipment, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, P.R. China.|Frank Reidy Center for Bioelectrics, Old Dominion University, Norfolk, VA, 23508, USA.|Department of Electrical and Computer Engineering, Old Dominion University, Norfolk, VA, 23529, USA.
Michael G
MG
Kong
D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D004247:DNA / Q000235:genetics*; D018274:Electroporation; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics*; D058626:Plasma Gases; D034741:RNA, Small Interfering / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D035683:MicroRNAs; D058626:Plasma Gases; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D004247:DNA