Dual-jet transition cold atmospheric plasma suppresses colon cancer stem cells via matrix metalloproteinase regulation

Colon cancer stem cells (CCSCs) are critical drivers of tumor progression, metastasis, and therapeutic resistance, largely mediated by matrix metalloproteinases (MMPs). This study investigates the therapeutic potential of a novel dual-jet transition cold atmospheric plasma (DJTCAP) system to modulateMMP expression and reduce CCSC viability. HT29-derived CCSCs were cultured in 3D Matrigel and exposed to three sequential plasma jets generated by a custom DJTCAP device, applied either via direct irradiation or plasma-activated medium for 120- or 240-seconds using helium or argon.MMP-1, -2, -3, -7, -9, and -14 expression was measured by RT-PCR, and cell viability was assessed using the MTT assay. DJTCAP treatment significantly downregulated allMMPs (p < 0.0001), with the most pronounced effects and cytotoxicity observed following 240-second helium exposure, which consistently outperformed argon. Plasma diagnostics confirmed efficient energy utilization (<1 W) and high generation of reactive oxygen and nitrogen species (RONS), suggesting a RONS-mediated mechanism of action. These results demonstrate that DJTCAP selectively suppressesMMP expression and reduces CCSC viability, highlighting its potential as a safe, energy-efficient, and targeted therapy for metastatic and treatment-resistant colorectal cancer.