{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:00:49Z","timestamp":1785427249806,"version":"3.56.0"},"reference-count":73,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"ODU Multidisciplinary Biomedical Research Seed Funding"},{"name":"Air Force Office of Scientific Research of the United States of America","award":["FA9550-22-1-0428"],"award-info":[{"award-number":["FA9550-22-1-0428"]}]},{"name":"U.S. Department of Energy, Office of Science, Office of Fusion Energy Sciences","award":["DE-SC0024627"],"award-info":[{"award-number":["DE-SC0024627"]}]},{"award":["DE-SC0024627"],"award-info":[{"award-number":["DE-SC0024627"]}],"id":[{"id":"https:\/\/ror.org\/02n3j8t12","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Pancreatic cancer remains a highly lethal malignancy, with standard therapies offering limited benefits in advanced stages; thus, novel strategies that exploit specific cancer cell vulnerabilities are urgently needed. Building on our previous findings that nanosecond pulsed electric fields (nsPEF) combined with cold atmospheric plasma (CAP) produce enhanced cytotoxicity, this study investigates the molecular mechanisms underlying this synergy. Pan02 pancreatic cancer cells were subjected to nsPEF, CAP, or a combination of both. We assessed cell viability, reactive oxygen species (ROS) production, and mitochondrial integrity using metabolic assays, flow cytometry, and fluorescence microscopy. Apoptotic markers were evaluated via Western blotting and caspase activity assays. Combined nsPEF\u2013CAP treatment significantly outperformed either modality alone in inducing cell death. Mechanistically, dual treatment triggered a surge in intracellular ROS, particularly mitochondrial superoxide, indicating severe oxidative stress. Distinct mitochondrial responses were observed: nsPEF reduced mitochondrial membrane potential, whereas CAP alone caused a slight elevation. Notably, while CAP induced apoptosis (evidenced by increased cleaved caspase-3 and caspase-3\/7 activity), lethal nsPEF (100 pulses) caused cell death without triggering apoptotic signaling. However, mild nsPEF (20 pulses) significantly potentiated CAP-induced apoptosis. These findings suggest that nsPEF sensitizes cells to CAP treatment by amplifying oxidative stress and mitochondrial dysfunction. This synergistic combination represents a promising therapeutic approach for managing pancreatic cancer cells resistant to conventional therapies.<\/jats:p>","DOI":"10.3390\/ijms27135933","type":"journal-article","created":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T10:41:01Z","timestamp":1782902461000},"page":"5933","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Synergistic Sensitization of Pancreatic Cancer Cells by Nanosecond Pulsed Electric Fields and Cold Atmospheric Plasma via Amplifying ROS and Apoptotic Signaling"],"prefix":"10.3390","volume":"27","author":[{"given":"Zobia","family":"Minhas","sequence":"first","affiliation":[{"name":"Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA 23508, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edwin A.","family":"Oshin","sequence":"additional","affiliation":[{"name":"Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA 23508, USA"},{"name":"Department of Electrical and Computer Engineering, Old Dominion University, Norfolk, VA 23529, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lifang","family":"Yang","sequence":"additional","affiliation":[{"name":"Leroy T. Canoles Jr. Cancer Research Center, Department of Biomedical and Translational Sciences, Macon & Joan Brock Virginial Health Sciences, Old Dominion University, Norfolk, VA 23501, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8895-2580","authenticated-orcid":false,"given":"Chunqi","family":"Jiang","sequence":"additional","affiliation":[{"name":"Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA 23508, USA"},{"name":"Department of Electrical and Computer Engineering, Old Dominion University, Norfolk, VA 23529, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7280-7888","authenticated-orcid":false,"given":"Siqi","family":"Guo","sequence":"additional","affiliation":[{"name":"Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA 23508, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1186\/s43046-025-00323-w","article-title":"Pancreatic cancer in the era of precision medicine: Challenges, advances, and the future of therapeutic strategies","volume":"37","author":"Silva","year":"2025","journal-title":"J. Egypt. Natl. Cancer Inst."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Zhu, H., Li, T., Du, Y., and Li, M. (2018). Pancreatic cancer: Challenges and opportunities. BMC Med., 16.","DOI":"10.1186\/s12916-018-1215-3"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1080\/14737140.2018.1417844","article-title":"The evolution into personalized therapies in pancreatic ductal adenocarcinoma: Challenges and opportunities","volume":"18","author":"Tesfaye","year":"2018","journal-title":"Expert Rev. Anticancer Ther."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1517\/14728222.2011.566216","article-title":"Overcoming drug resistance in pancreatic cancer","volume":"15","author":"Long","year":"2011","journal-title":"Expert Opin. Ther. Targets"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"E1085","DOI":"10.1055\/a-2379-1591","article-title":"Feasibility and safety of endoscopic ultrasound-guided diffusing alpha emitter radiation therapy for advanced pancreatic cancer: Preliminary data","volume":"12","author":"Miller","year":"2024","journal-title":"Endosc. Int. Open"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e530","DOI":"10.1097\/MPA.0000000000002489","article-title":"Factors Driving Pancreatic Cancer Survival Rates","volume":"54","author":"Rahib","year":"2025","journal-title":"Pancreas"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3217","DOI":"10.1200\/JCO.20.01364","article-title":"Metastatic pancreatic cancer: ASCO guideline update","volume":"38","author":"Sohal","year":"2020","journal-title":"J. Clin. Oncol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1089\/bioe.2024.0009","article-title":"Nano-Pulse Stimulation Therapy in Oncology","volume":"6","author":"Nuccitelli","year":"2024","journal-title":"Bioelectricity"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"McDaniel, A., Freimark, B., Navarro, C., Von Rothstein, K., Gonzalez, D., Linder, K., and Nuccitelli, R. (2023). Nano-pulse stimulation\u2122 therapy (NPS\u2122) is superior to cryoablation in clearing murine melanoma tumors. Front. Oncol., 12.","DOI":"10.3389\/fonc.2022.948472"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Pastori, C., Nafie, E.H., Wagh, M.S., Hunt, S.J., and Neal, R.E. (2024). Neoadjuvant chemo-immunotherapy is improved with a novel pulsed electric field technology in an immune-cold murine model. PLoS ONE, 19.","DOI":"10.1371\/journal.pone.0299499"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.15406\/jnmr.2015.02.00016","article-title":"Mechanisms of Nanosecond Pulsed Electric Field (NsPEF)-induced cell death in cells and tumors","volume":"2","author":"Beebe","year":"2015","journal-title":"J. Nanomed. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1016\/j.bbrc.2006.02.181","article-title":"Nanosecond pulsed electric fields cause melanomas to self-destruct","volume":"343","author":"Nuccitelli","year":"2006","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"23745","DOI":"10.1038\/s41598-021-03342-4","article-title":"The role of reactive oxygen species in the immunity induced by nano-pulse stimulation","volume":"11","author":"Guo","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Yao, C., Ma, X., Qian, K., Wang, Y., and Dong, S. (2023). Simulation and experimental study on the responses of subcellular structures in tumor cells induced by 5 ns pulsed electric fields. Appl. Sci., 13.","DOI":"10.3390\/app13148142"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Beebe, S.J., Chen, Y.-J., Sain, N.M., Schoenbach, K.H., and Xiao, S. (2012). Transient features in nanosecond pulsed electric fields differentially modulate mitochondria and viability. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0051349"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"136","DOI":"10.3390\/cells2010136","article-title":"Induction of cell death mechanisms and apoptosis by nanosecond pulsed electric fields (nsPEFs)","volume":"2","author":"Beebe","year":"2013","journal-title":"Cells"},{"key":"ref_17","first-page":"440","article-title":"Intracellular effect of ultrashort electrical pulses","volume":"22","author":"Schoenbach","year":"2001","journal-title":"Bioelectromagn. J. Bioelectromagn. Soc. Soc. Phys. Regul. Biol. Med. Eur. Bioelectromagn. Assoc."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1016\/j.bpj.2020.12.017","article-title":"2-ns Electrostimulation of Ca2+ influx into chromaffin cells: Rapid modulation by field reversal","volume":"120","author":"Zaklit","year":"2021","journal-title":"Biophys. J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1016\/j.bbamem.2012.11.032","article-title":"Primary pathways of intracellular Ca2+ mobilization by nanosecond pulsed electric field","volume":"1828","author":"Semenov","year":"2013","journal-title":"Biochim. Biophys. Acta (BBA)-Biomembr."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1259","DOI":"10.1007\/s10571-010-9573-1","article-title":"Nanosecond electric pulses: A novel stimulus for triggering Ca2+ influx into chromaffin cells via voltage-gated Ca2+ channels","volume":"30","author":"Craviso","year":"2010","journal-title":"Cell. Mol. Neurobiol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Guo, S., Burcus, N.I., Hornef, J., Jing, Y., Jiang, C., Heller, R., and Beebe, S.J. (2018). Nano-pulse stimulation for the treatment of pancreatic cancer and the changes in immune profile. Cancers, 10.","DOI":"10.3390\/cancers10070217"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zhao, J., Chen, S., Zhu, L., Zhang, L., Liu, J., Xu, D., Tian, G., and Jiang, T. (2021). Antitumor effect and immune response of nanosecond pulsed electric fields in pancreatic cancer. Front. Oncol., 10, Erratum in Front. Oncol.\u00a02022, 12, 1052763.","DOI":"10.3389\/fonc.2020.621092"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1038\/s41598-018-36527-5","article-title":"Mechanisms and immunogenicity of nsPEF-induced cell death in B16F10 melanoma tumors","volume":"9","author":"Rossi","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Xu, Z., Pan, C., Chen, L., Qian, J., Chen, X., Zhou, L., and Zheng, S. (2023). Nanosecond pulsed electric field induces an antitumor effect in triple-negative breast cancer via CXCL9 axis dependence in mice. Cancers, 15.","DOI":"10.3390\/cancers15072076"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Nuccitelli, R., Berridge, J.C., Mallon, Z., Kreis, M., Athos, B., and Nuccitelli, P. (2015). Nanoelectroablation of murine tumors triggers a CD8-dependent inhibition of secondary tumor growth. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0134364"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"108803","DOI":"10.1016\/j.bioelechem.2024.108803","article-title":"Anti-tumor effects of nanosecond pulsed electric fields in a murine model of pancreatic cancer","volume":"161","author":"Liang","year":"2025","journal-title":"Bioelectrochemistry"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1038\/s41598-023-27605-4","article-title":"Nanosecond pulsed electric field suppresses growth and reduces multi-drug resistance effect in pancreatic cancer","volume":"13","author":"Szlasa","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Yan, D., Malyavko, A., Wang, Q., Ostrikov, K., Sherman, J.H., and Keidar, M. (2021). Multi-modal biological destruction by cold atmospheric plasma: Capability and mechanism. Biomedicines, 9.","DOI":"10.3390\/biomedicines9091259"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"113513","DOI":"10.1063\/5.0020435","article-title":"Modulation of ROS in nanosecond-pulsed plasma-activated media for dosage-dependent cancer cell inactivation in vitro","volume":"27","author":"Jiang","year":"2020","journal-title":"Phys. Plasmas"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3317","DOI":"10.1109\/TPS.2021.3113260","article-title":"Synergistic effects of an atmospheric-pressure plasma jet and pulsed electric field on cells and skin","volume":"49","author":"Jiang","year":"2021","journal-title":"IEEE Trans. Plasma Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"885","DOI":"10.1038\/s41598-024-51298-y","article-title":"Synergistic effects of nanosecond pulsed plasma and electric field on inactivation of pancreatic cancer cells in vitro","volume":"14","author":"Oshin","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"116214","DOI":"10.1007\/s11356-023-30298-x","article-title":"Cold atmospheric plasma: A sustainable approach to inactivating viruses, bacteria, and protozoa with remediation of organic pollutants in river water and wastewater","volume":"30","author":"Hamza","year":"2023","journal-title":"Environ. Sci. Pollut. Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1002\/ppap.200700154","article-title":"Applied plasma medicine","volume":"5","author":"Fridman","year":"2008","journal-title":"Plasma Process. Polym."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Zhai, S.-y., Kong, M.G., and Xia, Y.-m. (2022). Cold atmospheric plasma ameliorates skin diseases involving reactive oxygen\/nitrogen species-mediated functions. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.868386"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1038\/s41419-022-04660-9","article-title":"Plasma-activated medium induces ferroptosis by depleting FSP1 in human lung cancer cells","volume":"13","author":"Jo","year":"2022","journal-title":"Cell Death Dis."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1038\/s41419-020-2459-3","article-title":"Cold atmospheric plasma induces GSDME-dependent pyroptotic signaling pathway via ROS generation in tumor cells","volume":"11","author":"Yang","year":"2020","journal-title":"Cell Death Dis."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Kniazeva, V., Tzerkovsky, D., Baysal, \u00d6., Kornev, A., Roslyakov, E., and Kostevitch, S. (2023). Adjuvant composite cold atmospheric plasma therapy increases antitumoral effect of doxorubicin hydrochloride. Front. Oncol., 13.","DOI":"10.3389\/fonc.2023.1171042"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Semmler, M.L., Bekeschus, S., Sch\u00e4fer, M., Bernhardt, T., Fischer, T., Witzke, K., Seebauer, C., Rebl, H., Grambow, E., and Vollmar, B. (2020). Molecular mechanisms of the efficacy of cold atmospheric pressure plasma (CAP) in cancer treatment. Cancers, 12.","DOI":"10.3390\/cancers12020269"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Min, T., Xie, X., Ren, K., Sun, T., Wang, H., Dang, C., and Zhang, H. (2022). Therapeutic effects of cold atmospheric plasma on solid tumor. Front. Med., 9.","DOI":"10.3389\/fmed.2022.884887"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Canady, J., Murthy, S.R., Zhuang, T., Gitelis, S., Nissan, A., Ly, L., Jones, O.Z., Cheng, X., Adileh, M., and Blank, A.T. (2023). The first cold atmospheric plasma phase I clinical trial for the treatment of advanced solid tumors: A novel treatment arm for cancer. Cancers, 15.","DOI":"10.3390\/cancers15143688"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"015208","DOI":"10.1088\/1361-6463\/50\/1\/015208","article-title":"Cold atmospheric plasma discharged in water and its potential use in cancer therapy","volume":"50","author":"Chen","year":"2016","journal-title":"J. Phys. D Appl. Phys."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Partecke, L.I., Evert, K., Haugk, J., Doering, F., Normann, L., Diedrich, S., Weiss, F.-U., Evert, M., Huebner, N.O., and Guenther, C. (2012). Tissue tolerable plasma (TTP) induces apoptosis in pancreatic cancer cells in vitro and in vivo. BMC Cancer, 12.","DOI":"10.1186\/1471-2407-12-473"},{"key":"ref_43","unstructured":"Honnorat, B. (2018). Application of Cold Plasma in Oncology, Multidisciplinary Experiments, Physical, Chemical and Biological Modeling, Sorbonne Universit\u00e9."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"15977","DOI":"10.18632\/oncotarget.13304","article-title":"Cold atmospheric plasma, a novel promising anti-cancer treatment modality","volume":"8","author":"Yan","year":"2017","journal-title":"Oncotarget"},{"key":"ref_45","unstructured":"Pouvesle, J.-M., Damany, X., Hamon, A., Valinattajomran, A., Marcelli, G., Stancampiano, A., Vijayarangan, V., Douat, C., Dozias, S., and Robert, E. (2018, January 1\u20135). Plasma\/target interactions in biomedical applications of cold atmospheric pressure plasmas. Proceedings of the PSE 2018, San Diego, CA, USA."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"103","DOI":"10.15344\/2456-8007\/2016\/103","article-title":"Clinical studies applying physical plasma in head and neck cancer-key points and study design","volume":"1","author":"Metelmann","year":"2016","journal-title":"Int. J. Clin. Res. Trials"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Wolff, C.M., Kolb, J.F., and Bekeschus, S. (2022). Combined in vitro toxicity and immunogenicity of cold plasma and pulsed electric fields. Biomedicines, 10.","DOI":"10.3390\/biomedicines10123084"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.abb.2012.08.004","article-title":"Oxidative effects of nanosecond pulsed electric field exposure in cells and cell-free media","volume":"527","author":"Pakhomova","year":"2012","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"580","DOI":"10.1016\/j.bbrc.2013.05.014","article-title":"Nanosecond pulsed electric field stimulation of reactive oxygen species in human pancreatic cancer cells is Ca2+-dependent","volume":"435","author":"Nuccitelli","year":"2013","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"e269","DOI":"10.1038\/emm.2016.119","article-title":"ROS homeostasis and metabolism: A critical liaison for cancer therapy","volume":"48","author":"Kim","year":"2016","journal-title":"Exp. Mol. Med."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"6907","DOI":"10.1038\/ncomms7907","article-title":"Amplification of oxidative stress by a dual stimuli-responsive hybrid drug enhances cancer cell death","volume":"6","author":"Noh","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Pourahmad, J., Salimi, A., and Seydi, E. (2016). Role of oxygen free radicals in cancer development and treatment. Free Radicals and Diseases, IntechOpen.","DOI":"10.5772\/64787"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1089\/ars.2011.4367","article-title":"The tumor microenvironment: Characterization, redox considerations, and novel approaches for reactive oxygen species-targeted gene therapy","volume":"19","author":"Policastro","year":"2013","journal-title":"Antioxid. Redox Signal."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Xu, D., Liu, D., Wang, B., Chen, C., Chen, Z., Li, D., Yang, Y., Chen, H., and Kong, M.G. (2015). In situ OH generation from O2\u2212 and H2O2 plays a critical role in plasma-induced cell death. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0128205"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"20332","DOI":"10.1038\/srep20332","article-title":"Cold atmospheric plasma jet-generated RONS and their selective effects on normal and carcinoma cells","volume":"6","author":"Kim","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1186\/s12964-024-01810-8","article-title":"Non-thermal atmospheric pressure plasma induces selective cancer cell apoptosis by modulating redox homeostasis","volume":"22","author":"Yun","year":"2024","journal-title":"Cell Commun. Signal."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"10048","DOI":"10.1038\/s41598-018-28443-5","article-title":"Cold atmospheric plasma causes a calcium influx in melanoma cells triggering CAP-induced senescence","volume":"8","author":"Schneider","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1002\/cbf.3823","article-title":"Cold atmospheric nitrogen plasma induces metal-initiated cell death by cell membrane rupture and mitochondrial perturbation","volume":"41","author":"Iuchi","year":"2023","journal-title":"Cell Biochem. Funct."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"13931","DOI":"10.1038\/s41598-019-50329-3","article-title":"Dynamics of singlet oxygen-triggered, RONS-based apoptosis induction after treatment of tumor cells with cold atmospheric plasma or plasma-activated medium","volume":"9","author":"Bauer","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"13818","DOI":"10.1038\/srep13818","article-title":"Electroporation of mammalian cells by nanosecond electric field oscillations and its inhibition by the electric field reversal","volume":"5","author":"Gianulis","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Pakhomova, O.N., Gregory, B.W., Semenov, I., and Pakhomov, A.G. (2013). Two modes of cell death caused by exposure to nanosecond pulsed electric field. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0070278"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"41163","DOI":"10.1038\/srep41163","article-title":"Mechanistic insights into the impact of Cold Atmospheric Pressure Plasma on human epithelial cell lines","volume":"7","author":"Dezest","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"17138","DOI":"10.1038\/s41598-019-53219-w","article-title":"Numerical modelling of the effects of cold atmospheric plasma on mitochondrial redox homeostasis and energy metabolism","volume":"9","author":"Murakami","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1016\/j.cell.2004.05.008","article-title":"Disruption of mitochondrial function during apoptosis is mediated by caspase cleavage of the p75 subunit of complex I of the electron transport chain","volume":"117","author":"Ricci","year":"2004","journal-title":"Cell"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"108568","DOI":"10.1016\/j.bioelechem.2023.108568","article-title":"Nanosecond pulsed electric fields (nsPEFs) modulate electron transport in the plasma membrane and the mitochondria","volume":"155","author":"Asadipour","year":"2024","journal-title":"Bioelectrochemistry"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"6288","DOI":"10.1021\/ja0588306","article-title":"Nanopore formation and phosphatidylserine externalization in a phospholipid bilayer at high transmembrane potential","volume":"128","author":"Vernier","year":"2006","journal-title":"J. Am. Chem. Soc."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Vernier, P.T., Sun, Y., and Gundersen, M.A. (2006). Nanoelectropulse-driven membrane perturbation and small molecule permeabilization. BMC Cell Biol., 7.","DOI":"10.1186\/1471-2121-7-37"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1016\/j.bpj.2012.11.3415","article-title":"Non-Ligand Mobilization of Intracellular Free Calcium by Nanosecond Pulsed Electric Field (NsPEF)","volume":"104","author":"Semenov","year":"2013","journal-title":"Biophys. J."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1111\/exd.12303","article-title":"First-in-human trial of nanoelectroablation therapy for basal cell carcinoma: Proof of method","volume":"23","author":"Nuccitelli","year":"2014","journal-title":"Exp. Dermatol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"4271065","DOI":"10.1155\/2017\/4271065","article-title":"Cold Atmospheric Plasma Induces Apoptosis and Oxidative Stress Pathway Regulation in T-Lymphoblastoid Leukemia Cells","volume":"2017","author":"Turrini","year":"2017","journal-title":"Oxid. Med. Cell. Longev."},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Wang, Y., Mang, X., Li, X., Cai, Z., and Tan, F. (2022). Cold atmospheric plasma induces apoptosis in human colon and lung cancer cells through modulating mitochondrial pathway. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.915785"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1016\/j.freeradbiomed.2018.10.434","article-title":"Roles of intracellular and extracellular ROS formation in apoptosis induced by cold atmospheric helium plasma and X-irradiation in the presence of sulfasalazine","volume":"129","author":"Rehman","year":"2018","journal-title":"Free Radic. Biol. Med."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"3289","DOI":"10.1097\/JS9.0000000000002361","article-title":"Nanosecond pulsed electric field ablation as first-line curative therapy for hepatocellular carcinoma in high-risk locations a prospective multicenter","volume":"111","author":"Xu","year":"2025","journal-title":"Int. J. Surg."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/13\/5933\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T10:50:22Z","timestamp":1782903022000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/13\/5933"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,1]]},"references-count":73,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["ijms27135933"],"URL":"https:\/\/doi.org\/10.3390\/ijms27135933","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,1]]}}}