Plasma-activated liquid (PAL) can effectively and selectively kill various types of cancer cells both in superficial and deeper tumors. As a promising novel approach to oncotherapy, the safety of PAL is essential in the clinic but has not been thoroughly assessed. In myeloma and blood tumors, the pathogenesis is in the bone marrow cavity. We have therefore evaluated the safety of PAL in New Zealand rabbits by intra-bone marrow injection, and provide a basis of further clinical research and application of PALs. In this study, both a plasma jet and plasma surface were used to treat saline solution, phosphate-buffered solution, and cell culture medium, to produce PAL. Then, oscillograms and optical emission spectra were evaluated to characterize the plasma discharge. Acute toxicity tests and safety evaluation studies were conducted by intra-bone marrow injection of PAL into New Zealand rabbits, while control rabbits received saline only. Body weight, vital organ coefficient, organ appearance, organ histopathology, blood cell and hemoglobin parameters, and blood biochemical indicators were tested on the 30th day after injection. We found that there was no mortality or loss of mobility throughout the experimental period. Acute toxicity tests showed that there were no PAL-related side effects in rabbits receiving the maximum dose of 700 μL PAL. PAL treatments did not affect body weight, organ coefficient, organ appearance, organ histopathology, or blood biochemical indicators. However, the percentage of lymphocytes decreased while the percentage of neutrophil granulocytes increased compared with the control group. In summary, our results indicate that PAL can be safely injected into bone marrow of New Zealand rabbits without significant toxicity.

Infection can hinder the process of wound healing, so it is important to begin antibacterial treatment quickly after a wound forms. Plasma activated water (PAW) can inactivate a variety of common wound infection bacteria. In this study, we compared the effects of PAW prepared with portable surface discharge plasma equipment and medical alcohol on wound healing in a mouse full-thickness skin wound model. The effectiveness of wound healing processes in mice was ranked accordingly: PAW treatment group > medical alcohol treatment group > control group. In order to further understand the mechanism of PAW in promoting wound healing, we tested the expression levels of the pro-inflammatory factors interleukin (IL)-1β and IL-6, the anti-inflammatory factor IL-10, and vascular endothelial growth factor (VEGF). The results showed that PAW promoted the release of pro-inflammatory factors and anti-inflammatory factors from the wounds in mice, which allowed the mice in the treatment group to transition out of the inflammatory period early and enter the next stage of wound healing. The expression level of VEGF in the wounds of mice in the PAW treatment group was higher, which indicates that the microvessels around the wound in the PAW treatment group proliferated faster, and thus the wound healed faster. PAW biosafety experiments showed that PAW did not significantly affect the appearance, morphology, or tissue structure of internal organs, or blood biochemical indicators in mice. In general, PAW prepared via portable devices is expected to become more widely used given its convenience, affordability, and lack of side effects in promoting wound healing.

In recent years, the emerging technology of cold atmospheric pressure plasma (CAP) has grown rapidly along with the many medical applications of cold plasma (e.g., cancer, skin disease, tissue repair, etc.). Plasma-activated liquids (e.g., culture media, water, or normal saline, previously exposed to plasma) are being studied as cancer treatments, and due to their advantages, many researchers prefer plasma-activated liquids as an alternative to CAP in the treatment of cancer. In this study, we showed that plasma-activated-saline (PAS) treatment significantly inhibited tumor growth, as compared with saline, in melanoma, and a low-pH environment had little effect on tumor growth in vivo. In addition, based on an ultra-high-performance liquid tandem chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) analysis of tumor cell metabolism, the glycerophospholipid metabolic pathway was the most susceptible metabolic pathway to PAS treatment in melanoma in vitro and in vivo. Furthermore, PAS also inhibited cell proliferation in vivo in oral tongue squamous-cell cancer and non-small-cell lung cancer. There were few toxic side effects in the three animal models, and the treatment was deemed safe to use. In the future, plasma-activated liquids may serve as a potential therapeutic approach in the treatment of cancer.

In this study, we investigated the effects of the quartz tube diameter, air flow rate, and applied voltage on the characteristics of an air plasma jet to obtain the optimized discharge characteristics. The physicochemical properties and concentration of reactive oxygen and nitrogen species (RONS) in plasma-activated medium (PAM) were characterized to explore their chemical activity. Furthermore, we investigated the inactivation effect of air plasma jet on tumour cells and their corresponding inactivation mechanism. The results show that the tube diameter plays an important role in sustaining the voltage of the air plasma jet, and the gas flow rate affects the jet length and discharge intensity. Additionally, the air plasma jet discharge displays two modes, namely, ozone and nitrogen oxide modes at high and low gas flow rates, respectively. Increasing the voltage increases the concentration of reactive species and the length of discharge. By evaluating the viability of A549 cells under different parameters, the optimal treatment conditions were determined to be a quartz tube diameter of 4 mm, gas flow rate of 0.5 SLM, and voltage of 18 kV. Furthermore, an air plasma jet under the optimized conditions effectively enhanced the chemical activity in PAM and produced more aqueous RONS. The air plasma jet induced significant cytotoxicity in A549 cancer cells after plasma treatment. H 2 O 2 and -NO 2 are regarded as key factors in promoting cell inactivation. The present study demonstrates the potential use of tumour cell therapy by atmospheric air PAM, which aids a better understanding of plasma liquid chemistry.

In the field of plasma biomedicine, research on a plasma-activated medium (PAM) has attracted increasing attention in recent years because of its excellent characteristics. In this study, we used an atmospheric pressure air plasma jet to treat four different solutions: de-ionized water, RPMI 1640 medium, phosphate buffered saline (PBS), and saline. In order to investigate the property differences of different PAM, we mainly analyzed the physical and chemical properties and liquid-phase active species of different PAM and evaluated the inactivation of A549 lung cancer cells. The results show that the concentrations of long-lived reactive species (H2O2, NO2−, and NO3−) in different PAM increased with increasing treatment time. Biological experiments showed that the antitumor effects were in the order of PBS > saline > RPMI 1640 medium, and the best inactivation effect of plasma-activated PBS for 12 min was 89%. Meanwhile, plasma-activated PBS effectively promoted apoptosis in A549 cells, and the highest apoptosis rate was 91.3%. Therefore, this study demonstrates the medical application of different PAM in killing cancer cells and promotes the understanding of plasma–liquid interaction.