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H2O2-mediated Redox Priming by Non-thermal Microplasma Enhances Macromolecule Delivery to HL-60 Cells via Endocytosis

delete2026-06-12
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M
Mahedi Hasan *
J
Jaroslav Kristof
M
Md. Jahangir Alam
F
Farhana Begum
R
Rubaiya Islam
K
Kazuo Shimizu *
DOI:10.1007/s11090-026-10682-4delete
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Abstract

Abstract

En 中文
Efficient delivery of therapeutic macromolecules into hematopoietic suspension cells remains a bottleneck for gene- and immune-therapy workflows. Here we frame a contact-free microplasma priming approach as ReMod (Redox-guided Modulation): a reversible, cytocompatible “tune-rather-than-force” strategy that gates uptake through controlled extracellular redox inputs. A 150-kDa fluorescein isothiocyanate–dextran (FD-150) was used as a model cargo in HL-60 leukemia cells. Culture-medium RONS were tuned using N2 + 0–10% O2 + H2O, Ar + 0–10% O2 + H2O, and Ar + 0–10% N2 + H2O, all at a total flow of 4 Lmin⁻1. A brief 3-min non-thermal exposure produced a chemically tunable plasma–liquid interface and revealed a cytocompatible RONS “sweet spot” for FD-150 internalization, maximized under N2 + 5% O2 + H2O. Scavenger experiments, together with exogenous H2O2, indicated that extracellular H2O2 is a principal effector of uptake. Time-integrated intracellular ROS showed a modest O2-dependent increase, whereas NO-related fluorescence exhibited a larger fold-increase at 1–5% O2, suggesting that NO signaling may act as a later, reinforcing component of the redox response. qRT-PCR profiling of endocytic markers was consistent with clathrin-mediated endocytosis and macropinocytosis as dominant uptake routes under ReMod priming. These findings define a practical redox-gating window for macromolecule delivery into suspension leukemia cells without electroporation and provide a basis for extending microplasma ReMod to nucleic-acid or immunotherapeutic payloads.
Keywords:
Microplasma
ReMod (Redox-guided Modulation)
Redox priming
RONS
HL-60
FITC-dextran
Macromolecule delivery
Clathrin-mediated endocytosis
Macropinocytosis
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Journal

Plasma Chemistry and Plasma Processing cover
Plasma Chemistry and Plasma Processing
IF:
2.5
Papers:
175
Citations:
4.1K

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G
graduate school of medical photonics
Scholars:
7
Papers: 2
Citations: 0
G
Graduate School of Science and Technology
Scholars:
135
Papers: 72
Citations: 1
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