Background: Glioblastoma (GBM) resists current therapies due to its rapid proliferation, diffuse invasion, and heterogeneous cell populations. We previously showed that a single cold atmospheric plasma discharge tube (DT) reduces GBM viability via broad-spectrum electromagnetic (EM) emissions. Here, we tested whether two DTs arranged in a helmet configuration could generate overlapping EM fields to amplify the anti-tumor effects without thermal injury. Methods: The physical outputs of the single- and dual-DT setups were characterized by infrared thermography, broadband EM field probes, and oscilloscope analysis. Human U87-MG cells were exposed under the single or dual configurations. The viability was quantified with WST-8 assays mapped across 96-well plates; the intracellular reactive oxygen species (ROS), membrane integrity, apoptosis, and mitochondrial potential were assessed by multiparametric flow cytometry. Our additivity models compared the predicted versus observed dual-DT cytotoxicity. Results: The dual-DT operation produced constructive EM interference, elevating electric and magnetic field amplitudes over a broader area than either tube alone, while temperatures remained <39 °C. The single-DT exposure lowered the cell viability by ~40%; the dual-DT treatment reduced the viability by ~60%, exceeding the additive predictions. The regions of greatest cytotoxicity co-localized with the zones of highest EM field overlap. The dual-DT exposure doubled the intracellular ROS compared with single-DT and Annexin V positivity, confirming oxidative stress-driven cell death. The out-of-phase operation of the discharge tubes enabled the localized control of the treatment regions, which can guide future treatment planning. Conclusions: Two synchronously operated plasma discharge tubes synergistically enhanced GBM cell killing through non-thermal mechanisms that coupled intensified overlapping EM fields with elevated oxidative stress. This positions modular multi-DT arrays as a potential non-invasive adjunct or alternative to existing electric-field-based therapies for glioblastoma.
背景：胶质母细胞瘤（GBM）因其快速增殖、弥漫性侵袭及异质性细胞群而对现有疗法产生抵抗。我们先前研究表明，单根冷大气等离子体放电管（DT）可通过广谱电磁（EM）发射降低GBM细胞活性。本研究旨在探讨以头盔式排列的双放电管能否通过重叠电磁场增强抗肿瘤效应，同时避免热损伤。方法：通过红外热成像、宽带电磁场探头及示波器分析，对单管与双管装置的物理输出特性进行表征。将人源U87-MG细胞分别置于单管或双管配置下进行暴露处理。采用WST-8法对96孔板内细胞活性进行空间分布定量分析；通过多参数流式细胞术检测细胞内活性氧（ROS）水平、膜完整性、细胞凋亡及线粒体膜电位。通过加和性模型比较双管细胞毒性的预测值与观测值。结果：双管运行产生相长电磁干涉，在更广区域内提升电场与磁场振幅，同时温度始终低于39°C。单管暴露使细胞活性降低约40%，而双管处理使活性降低约60%，显著超过加和预测值。最大细胞毒性区域与电磁场重叠最显著区域高度重合。与单管相比，双管暴露使细胞内ROS水平倍增，Annexin V阳性率升高，证实氧化应激驱动的细胞死亡。放电管的异相运行为治疗区域的局部调控提供了可能，可为未来治疗规划提供指导。结论：双管同步运行通过非热机制协同增强GBM细胞杀伤效果，该机制耦合了增强的重叠电磁场与升高的氧化应激水平。这表明模块化多管阵列有望成为胶质母细胞瘤现有电场疗法的潜在无创辅助或替代方案。
Dual-Plasma Discharge Tube for Synergistic Glioblastoma Treatment
肿瘤(癌症)患者之家