High-intensity pulsed electromagnetic fields (HI-PEMF) can be used to trigger contactless permeabilization of the plasma membrane similar to electroporation (EP). The permeabilization efficiency and gene delivery by HI-PEMF in vitro are currently inferior to EP. It was suggested that the methodology can be improved with conductive gold nanoparticles (AuNPs), which are reported to amplify the induced electric field in close proximity to the cell membrane.
Objectives:
Therefore, in this work, we have studied different NPs, which varied in material/conductivity (gold and silica), size (10–50+ nm), shape (i.e., round and rods), concentration (50–200 µg/mL), and functionalization (pegylated or not), and combined them with HI-PEMF (6.7 T × 100 pulses, 1 Hz).
Methods:
The normal Chinese hamster ovary cell line (CHO) and the cancer human urinary bladder’s transitional carcinoma cell line (T24) were used as a model. We have characterized cell membrane permeabilization using propidium iodide (PI) and the efficacy of gene delivery using pEGFP-N1.
Results:
Larger NPs and higher NP concentrations resulted in up to a 10% increase in membrane permeability. In contrast, semispherical and rod-shaped AuNPs did not further enhance permeabilization efficiency. Gene delivery efficiency increased from 3% in control samples to 6% in the presence of 50 nm AuNPs. Overall, CHO cells were more susceptible to HI-PEMF-induced effects than T24 cells.
Conclusions:
This study shows the potential to increase gene delivery efficacy by combining HI-PEMF treatment with conductive NPs. However, it was concluded that the HI-PEMF-induced effects are highly dependent on the cell line, NP type, and concentration and therefore require further investigation.
GinnSL, AmayaAK, AlexanderIE, et al.Gene therapy clinical trials worldwide to 2017: An update. J Gene Med, 2018; 20(5):e3015–e3016; doi: 10.1002/jgm.3015
2.
BroderickKE, ChanA, LinF, et al.Optimized in vivo transfer of small interfering RNA targeting dermal tissue using in vivo surface electroporation. Mol Ther Nucleic Acids, 2012; 1(2):e11; doi: 10.1038/mtna.2012.1
3.
SachdevS, PotočnikT, RemsL, et al.Revisiting the role of pulsed electric fields in overcoming the barriers to in vivo gene electrotransfer. Bioelectrochemistry, 2022; 144:107994; doi: 10.1016/J.BIOELECHEM.2021.107994
4.
RosazzaC, MeglicHS, ZumbuschA, et al.Gene electrotransfer: A mechanistic perspective. Curr Gene Ther, 2016; 16(2):98–129; doi: 10.2174/1566523216666160331130040
5.
HernándezJL, CollT, CiudadCJ. A highly efficient electroporation method for the transfection of endothelial cells. Angiogenesis, 2004; 7(3):235–241; doi: 10.1007/s10456-004-4180-8
6.
MashelTV, TarakanchikovaYV, MuslimovAR, et al.Biomaterials Overcoming the delivery problem for therapeutic genome editing : Current status and perspective of non-viral methods. Biomaterials, 2020; 258(July):120282; doi: 10.1016/j.biomaterials.2020.120282
7.
GeboersB, SchefferHJ, GraybillPM, et al.High-Voltage electrical pulses in oncology: Irreversible electroporation, electrochemotherapy, gene electrotransfer, electrofusion, and electroimmunotherapy. Radiology, 2020; 295(2):254–272; doi: 10.1148/radiol.2020192190
8.
BradleyCJ, HainesDE. Pulsed field ablation for pulmonary vein isolation in the treatment of atrial fibrillation. J Cardiovasc Electrophysiol, 2020; 31(8):2136–2147.
9.
ChunK-RJ, MiklavčičD, VlachosK, et al.State-of-the-Art pulsed field ablation for cardiac arrhythmias : Ongoing evolution and future perspective. Europace, 2024; 26(6):euae134; doi: 10.1093/europace/euae134
NovickijV, DermolJ, GrainysA, et al.Membrane permeabilization of mammalian cells using bursts of high magnetic field pulses. PeerJ, 2017; 5(April):e3267; doi: 10.7717/peerj.3267
12.
KranjcS, KranjcM, ScancarJ, et al.Electrochemotherapy by pulsed electromagnetic field treatment (PEMF) in mouse melanoma B16F10 in vivo. Radiol Oncol, 2016; 50(1):39–48; doi: 10.1515/raon-2016-0014
13.
NovickijV, KranjcM, StaigvilaG, et al.High-Pulsed electromagnetic field generator for contactless permeabilization of cells in vitro. IEEE Trans Magn, 2020; 56(5):1–6; doi: 10.1109/TMAG.2020.2979120
14.
ChenC, EvansJA, RobinsonMP, et al.Electroporation of cells using EM induction of ac fields by a magnetic stimulator. Phys Med Biol, 2010; 55(4):1219–1229; doi: 10.1088/0031-9155/55/4/021
15.
Kranjc BrezarS, KranjcM, ČemažarM, et al.Electrotransfer of siRNA to silence enhanced green fluorescent protein in tumor mediated by a high intensity pulsed electromagnetic field. Vaccines (Basel), 2020; 8(1):49; doi: 10.3390/vaccines8010049
16.
MiklavcicD, NovickijV, KranjcM, et al.Contactless electroporation induced by high intensity pulsed electromagnetic fields via distributed nanoelectrodes. Bioelectrochemistry, 2020; 132:107440; doi: 10.1016/j.bioelechem.2019.107440
17.
KranjcM, Dermol-ČerneJ, PotočnikT, et al.High-Intensity pulsed electromagnetic field-mediated gene electrotransfection in vitro. Int J Mol Sci, 2022; 23(17):9543; doi: 10.3390/ijms23179543
18.
GoršakT, DrabM, KrižajD, et al.Magneto-mechanical actuation of barium-hexaferrite nanoplatelets for the disruption of phospholipid membranes. J Colloid Interface Sci, 2020; 579:508–519; doi: 10.1016/j.jcis.2020.06.079
19.
GhorbelA, AndréFM, MirLM, et al.Electrophoresis-assisted accumulation of conductive nanoparticles for the enhancement of cell electropermeabilization. Bioelectrochemistry, 2021; 137:107642; doi: 10.1016/j.bioelechem.2020.107642
20.
RolongA, ProkopKJ, DavalosRV. Impact of the use of nanoparticles on electric field distribution during irreversible electroporation treatments: Can the lesion be enhanced beyond IRE margin? In: IFMBE Proceedings, 2015:793–796; doi: 10.1007/978-3-319-11128-5_197
21.
RaffaV, RiggioC, SmithMW, et al.BNNT-Mediated irreversible electroporation: Its potential on cancer cells. Technol Cancer Res Treat, 2012; 11(5):459–465; doi: 10.7785/tcrt.2012.500258
22.
ZuY, HuangS, LiaoWC, et al.Gold nanoparticles enhanced electroporation for mammalian cell transfection. J Biomed Nanotechnol, 2014; 10(6):982–992; doi: 10.1166/jbn.2014.1797
RezaeeZ, YadollahpourA, BayatiV, et al.Gold nanoparticles and electroporation impose both separate and synergistic radiosensitizing effects in HT-29 tumor cells: An in vitro study. Int J Nanomedicine, 2017; 12:1431–1439; doi: 10.2147/IJN.S128996
25.
NikoobakhtB, El-SayedMA. Preparation and growth mechanism of Gold Nanorods (NRs) using seed-mediated growth method. Chem Mater, 2003; 15(10):1957–1962; doi: 10.1021/cm020732l
Ojea-JiménezI, BastúsNG, PuntesV. Influence of the sequence of the reagents addition in the citrate-mediated synthesis of gold nanoparticles. J Phys Chem C, 2011; 115(32):15752–15757; doi: 10.1021/jp2017242
28.
BastúsNG, ComengeJ, PuntesV. Kinetically controlled seeded growth synthesis of citrate-stabilized gold nanoparticles of up to 200 nm: Size focusing versus ostwald ripening. Langmuir, 2011; 27(17):11098–11105; doi: 10.1021/la201938u
29.
StöberW, FinkA, BohnE. Controlled growth of monodisperse silica spheres in the micron size range. J Colloid Interface Sci, 1968; 26(1):62–69; doi: 10.1016/0021-9797(68)90272-5
30.
HanahanD, WeinbergRA. Hallmarks of cancer: The next generation. Cell, 2011; 144(5):646–674; doi: 10.1016/j.cell.2011.02.013
31.
DermolJ, PakhomovaON, PakhomovAG, et al.Cell electrosensitization exists only in certain electroporation buffers. PLoS One, 2016; 11(7):e0159434; doi: 10.1371/journal.pone.0159434
32.
PolajžerT, Dermol-ČerneJ, ReberšekM, et al.Cancellation effect is present in high-frequency reversible and irreversible electroporation. Bioelectrochemistry, 2020; 132:107442; doi: 10.1016/J.BIOELECHEM.2019.107442
33.
PolajžerT, KranjcM, KraljS, et al.Limited efficacy of nanoparticle-assisted electroporation for membrane permeabilization and gene electrotransfer. Pharmaceutics, 2025; 17(8):964; doi: 10.3390/pharmaceutics17080964
34.
Franco-UlloaS, TatulliG, BoreSL, et al.Dispersion state phase diagram of citrate-coated metallic nanoparticles in saline solutions. Nat Commun, 2020; 11(1):5422; doi: 10.1038/s41467-020-19164-3
35.
ParkJ-W, Shumaker-ParryJS. Structural study of citrate layers on gold nanoparticles: Role of intermolecular interactions in stabilizing nanoparticles. J Am Chem Soc, 2014; 136(5):1907–1921; doi: 10.1021/ja4097384
36.
WuX, DaiH, LiuL, et al.Citrate reduced oxidative damage in stem cells by regulating cellular redox signaling pathways and represent a potential treatment for oxidative stress-induced diseases. Redox Biol, 2019; 21:101057; doi: 10.1016/j.redox.2018.11.015
37.
CaiazzaC, D’AgostinoM, PassaroF, et al.Effects of long-term citrate treatment in the PC3 prostate cancer cell line. Int J Mol Sci, 2019; 20(11):2613; doi: 10.3390/ijms20112613
38.
von SchirndingC, GiopanouI, HermawanA, et al.Synergistic combination of calcium and citrate in mesoporous nanoparticles targets pleural tumors. Chem, 2021; 7(2):480–494; doi: 10.1016/j.chempr.2020.11.021
39.
CostelloLC, FranklinRB. The implications of the hypocitricemic response to surgery and the role of liver function and hepatocyte metabolism: An important, but neglected, clinical relationship. J Liver Res Disord Ther, 2018; 4(3):114–119; doi: 10.15406/jlrdt.2018.04.00112
40.
SukJS, XuQ, KimN, et al.PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Adv Drug Deliv Rev, 2016; 99(Pt A):28–51; doi: 10.1016/j.addr.2015.09.012
41.
RosazzaC, DeschoutH, BuntzA, et al.Endocytosis and endosomal trafficking of DNA after gene electrotransfer in vitro. Mol Ther Nucleic Acids, 2016; 5(2):e286; doi: 10.1038/mtna.2015.59
42.
WuM, YuanF. Membrane binding of plasmid DNA and endocytic pathways are involved in electrotransfection of mammalian cells. PLoS One, 2011; 6(6):e20923; doi: 10.1371/journal.pone.0020923
43.
PotočnikT, SachdevS, PolajžerT, et al.Efficient gene transfection by electroporation—In vitro and in silico study of pulse parameters. Appl Sci, 2022; 12(16):8237; doi: 10.3390/app12168237
