Canady Helios Cold Plasma: The First Non-Thermal (240 C), Non-Contact, Irreversible Electroporation (NTNC-IRETM), Plasma Treated Electromagnetic Field (PTEFTM) Induces Cell Death in Breast Cancer and Tumor Cells at the Surgical Margin

Canady Helios Cold Plasma (CHCP) system has emerged as a promising modality for cancer therapy, offering selective tumor cell death while sparing surrounding healthy tissue. The CHCP system, a Cold atmospheric plasma (CAP) technology, creates reactive oxygen and nitrogen species via plasma treated electromagnetic fields (PTEFTM) producing irreversible electroporation (IRE). In this study, we evaluated CHCP-induced IRE across four biologically distinct breast cancer cell lines: triple-negative (MDA-MB-231 and Hs578T), ER⁺/PR⁺/HER2⁻ (MCF-7), and ER⁺/PR⁺/HER2⁺ (BT-474). Membrane permeability was quantified via propidium iodide (PI) uptake following 5-minute exposures at 25 V (~1675 V/cm PTEF) and 30 V (~2010 V/cm). CHCP induced voltage and time dependent IRE at 30 V produced robust and sustained PI uptake across all lines, compared to 25 V at temperatures ranging from 22-240 C. Morphological analyses revealed membrane blebbing, cell shrinkage, and detachment consistent with IRE and oxidative stress. Functional studies demonstrated CHCP-mediated intracellular delivery of siRNA targeting BCL2A1 and suppression of clonogenic potential, confirming pore formation sufficient for macromolecular transport. Ex vivo treatment of tumor margins from our Phase I clinical trial (NCT04267575) specimens corroborated selective tumor cell death while preserving non-cancerous cell integrity. The authors are the first to demonstrate CHCP as a non-thermal (240 C), non-contact plasma-based IRE platform which induce controlled membrane permeabilization and selective cancer cell death. CHCP offers a translational strategy to eradicate residual tumor cells at the surgical margins, and prevent local recurrence, positioning it as a versatile adjunct in precision surgical oncology.