The efficacy of helium (He) and argon (Ar) plasma jets are being investigated for different healthcare applications including wound and cancer therapy, sterilisation and surface disinfections. Current research points to a potential link between the generation of reactive oxygen and nitrogen species (RONS) and outcomes in a range of biological and medical applications. As new data accrue, further strengthening this link, it becomes important to understand the controlled delivery of RONS into solutions, tissue fluids and tissues. This paper investigates the use of He and Ar plasma jets to deliver three RONS (hydrogen peroxide—H ₂ O ₂ , nitrite— NO 2 − and nitrate— NO 3 − ) and molecular oxygen (O ₂ ) directly into deionised (DI) water, or indirectly into DI water through an agarose target. The DI water is used in place of tissue fluid and the agarose target serves as a surrogate of tissue. Direct plasma jet treatments deliver more RONS and O ₂ than the through-agarose treatments for equivalent treatments times. The former only deliver RONS whilst the plasma jets are ignited; the latter continues to deliver RONS into the DI water long after the plasmas are extinguished. The He plasma jet is more effective at delivering H ₂ O ₂ and NO 2 − directly into DI water, but the Ar plasma jet is more effective at nitrating the DI water in both direct and through-agarose treatments. DI water directly treated with the plasma jets is deoxygenated, with the He plasma jet purging more O ₂ than the Ar plasma jet. This effect is known as ‘sparging’. In contrast, for through-agarose treatments both jets oxygenated the DI water. These results indicate that in the context of direct and indirect plasma jet treatments of real tissue fluids and tissue, the choice of process gas (He or Ar) could have a profound effect on the concentrations of RONS and O ₂ . Irrespective of operating gas, sparging of tissue fluid (in an open wound) for long prolonged periods during direct plasma jet treatment, could have implications for healthy tissue function; whilst through-tissue plasma jet treatment may provide a method to reperfuse oxygen-starved tissue. The assays described in this paper can be readily adopted (by others) and may support the future development of plasma sources to deliver specific (metered) doses of RONS.

We explore how to configure an argon atmospheric-pressure plasma jet for enhancing its production of hydrogen peroxide (H₂O₂) in deionised water (DIW). The plasma jet consists of a quartz tube of 1.5 mm inner diameter and 3 mm outer diameter, with an upstream internal needle electrode (within the tube) and a downstream external cylindrical electrode (surrounding the tube). The plasma is operated by purging argon through the glass tube and applying a sinusoidal AC voltage to the internal needle electrode at 10 kV (peak–peak) with a frequency of 23.5 kHz. We study how the following operational parameters influence the production rate of H₂O₂ in water: tube length, inter-electrode separation distance, distance of the ground electrode from the tube orifice, distance between tube orifice and the DIW, argon flow rate and treatment time. By examining the electrical and optical properties of the plasma jet, we determine how the above operational parameters influence the major plasma processes that promote H₂O₂ generation through electron-induced dissociation reactions and UV photolysis within the plasma core and in the plasma afterglow; but with a caveat being that these processes are highly dependent on the water vapour content from the argon gas supply and ambient environment. We then demonstrate how the synergistic action between H₂O₂ and other plasma generated molecules at a plasma induced low pH in the DIW is highly effective at decontaminating common wound pathogens Gram-positive Staphylococus aureus and Gram-negative Pseudomonas aeruginosa. The information presented in this study is relevant in the design of medical plasma devices where production of plasma reactive species such as H₂O₂ at physiologically useful concentrations is needed to help realise the full clinical potential of the technology.

Cold atmospheric plasmas have attracted significant worldwide attention for their potential beneficial effects in cancer therapy. In order to further improve the effectiveness of plasma in cancer therapy, it is important to understand the generation and transport of plasma reactive species into tissue fluids, tissues and cells, and moreover the rates and depths of delivery, particularly across physical barriers such as skin. In this study, helium (He) plasma jet treatment of a 3D cancer tumour, grown on the back of a live mouse, induced apoptosis within the tumour to a depth of 2.8 mm. The He plasma jet was shown to deliver reactive oxygen species through the unbroken skin barrier before penetrating through the entire depth of the tumour. The depth and rate of transport of He plasma jet generated H ₂ O ₂ , NO ₃ ⁻ and NO ₂ ⁻ , as well as aqueous oxygen [O ₂ (aq)], was then tracked in an agarose tissue model. This provided an approximation of the H ₂ O ₂ , NO ₃ ⁻ , NO ₂ ⁻ and O ₂ (aq) concentrations that might have been generated during the He plasma jet treatment of the 3D tumour. It is proposed that the He plasma jet can induce apoptosis within a tumour by the ‘deep’ delivery of H ₂ O ₂ , NO ₃ ⁻ and NO ₂ ⁻ coupled with O ₂ (aq); the latter raising oxygen tension in hypoxic tissue.

The influence of protein and molecular, ground state oxygen (O₂) on the plasma generation, and transport of reactive oxygen and nitrogen species (RONS) in tissue are investigated. A tissue target, comprising a 1 mm thick gelatin film (a surrogate for real tissue), is placed on top of a 96-well plate; each well is filled with phosphate buffered saline (PBS, pH 7.4) containing one fluorescent or colorimetric reporter that is specific for one of three RONS (i.e., H₂O₂, NO₂⁻, or OH^•) or a broad spectrum reactive oxygen species reporter (2,7-dichlorodihydrofluorescein). A helium cold atmospheric plasma (CAP) jet contacts the top of the gelatin surface, and the concentrations of RONS generated in PBS are measured on a microplate reader. The data show that H₂O₂, NO₂⁻, or OH^• are generated in PBS underneath the target. Independently, measurements are made of the O₂ concentration in the PBS with and without the gelatin target. Adding bovine serum albumin protein to the PBS or gelatin shows that protein either raises or inhibits RONS depending upon the O₂ concentration. Our results are discussed in the context of plasma-soft tissue interactions that are important in the development of CAP technology for medicine, biology, and food manufacturing.