title: Helium generated cold plasma finely regulates activation of human fibroblast-like primary cells.
abstract: Non-thermal atmospheric pressure plasmas are being developed for a wide range of health care applications, including wound healing. However in order to exploit the potential of plasma for clinical applications, the understanding of the mechanisms involved in plasma-induced activation of fibroblasts, the cells active in the healing process, is mandatory. In this study, the role of helium generated plasma in the tissue repairing process was investigated in cultured human fibroblast-like primary cells, and specifically in hepatic stellate cells and intestinal subepithelial myofibroblasts. Five minutes after treatment, plasma induced formation of reactive oxygen species (ROS) in cultured cells, as assessed by flow cytometric analysis of fluorescence-activated 2',7'-dichlorofluorescein diacetate probe. Plasma-induced intracellular ROS were characterized by lower concentrations and shorter half-lives with respect to hydrogen peroxide-induced ROS. Moreover ROS generated by plasma treatment increased the expression of peroxisome proliferator activated receptor (PPAR)-γ, nuclear receptor that modulates the inflammatory responses. Plasma exposure promoted wound healing in an in vitro model and induced fibroblast migration and proliferation, as demonstrated, respectively, by trans-well assay and partitioning between daughter cells of carboxyfluorescein diacetate succinimidyl ester fluorescent dye. Plasma-induced fibroblast migration and proliferation were found to be ROS-dependent as cellular incubation with antioxidant agents (e.g. N-acetyl L-cysteine) cancelled the biological effects. This study provides evidence that helium generated plasma promotes proliferation and migration in liver and intestinal fibroblast-like primary cells mainly by increasing intracellular ROS levels. Since plasma-evoked ROS are time-restricted and elicit the PPAR-γ anti-inflammatory molecular pathway, this strategy ensures precise regulation of human fibroblast activation and can be considered a valid therapeutic approach for liver and gut lesions.
authors.affiliation: Department of Molecular Medicine, Unit of Microbiology, University of Padova, Padova, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Brun
authors.affiliation: Department of Molecular Medicine, Unit of Microbiology, University of Padova, Padova, Italy; Department of Surgical, Oncological and Gastroenterological Sciences, University of Padova, Padova, Italy.
authors.forename: Surajit
authors.initials: S
authors.lastname: Pathak
authors.affiliation: Department of Molecular Medicine, Unit of Microbiology, University of Padova, Padova, Italy.
authors.forename: Ignazio
authors.initials: I
authors.lastname: Castagliuolo
authors.affiliation: Department of Molecular Medicine, Unit of Microbiology, University of Padova, Padova, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Palù
authors.affiliation: Department of Molecular Medicine, Unit of Histology, University of Padova, Padova, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Brun
authors.affiliation: Consorzio RFX, Padova, Italy; Istituto Gas Ionizzati del CNR, Padova, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Zuin
authors.affiliation: Consorzio RFX, Padova, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Cavazzana
authors.affiliation: Consorzio RFX, Padova, Italy; Istituto Gas Ionizzati del CNR, Padova, Italy.
authors.forename: Emilio
authors.initials: E
authors.lastname: Martines
mesh_terms: D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism*; D006371:Helium; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D047495:PPAR gamma / Q000378:metabolism; D058626:Plasma Gases / Q000494:pharmacology*; D061251:Primary Cell Culture; D017382:Reactive Oxygen Species / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects
chemical_list: D016207:Cytokines; D047495:PPAR gamma; D058626:Plasma Gases; D017382:Reactive Oxygen Species; D006371:Helium