Non-thermal atmospheric pressure plasma has been proposed as a new tool for various biological and medical applications. Plasma in close proximity to cell culture media or water creates reactive oxygen and nitrogen species containing solutions known as plasma-activated media (PAM) or plasma-activated water (PAW) – the latter even displays acidification. These plasma-treated solutions remain stable for several days with respect to the storage temperature. Recently, PAM and PAW have been widely studied for many biomedical applications. Here, we reviewed promising reports demonstrating plasma-liquid interaction chemistry and the application of PAM or PAW as an anti-cancer, anti-metastatic, antimicrobial, regenerative medicine for blood coagulation and even as a dental treatment agent. We also discuss the role of PAM on cancer initiation cells (spheroids or cancer stem cells), on the epithelial mesenchymal transition (EMT), and when used for metastasis inhibition considering its anticancer effects. The roles of PAW in controlling plant disease, seed decontamination, seed germination and plant growth are also considered in this review. Finally, we emphasize the future prospects of PAM, PAW or plasma-activated solutions in biomedical applications with a discussion of the mechanisms and the stability and safety issues in relation to humans.

Atmospheric-pressure non-thermal bio-compatible plasma is a partially ionized gas with electrically charged particles. Previous studies demonstrated that dielectric barrier discharge (DBD) plasma could induce apoptosis of various cancer cells, in particular demonstrating the selective cytotoxicity of cancer cells over normal cells. Therefore, DBD plasma can be considered as a potential cancer treatment method for clinical applications. We previously developed a microwave jet plasma system, producing nitric oxide called nitric oxide-plasma activated water (NO-PAW). In this study, we explored the effects of NO-PAW on a cervical cancer cell line, in comparison with DBD plasma. The cytotoxicity results showed that the treatment of HeLa cell with DBD for 4 minutes and 7 μM concentration of NO-PAW could reach almost IC₆₀. For the apoptosis assay, 4 minutes treatment of DBD could induce 7% apoptotic effect, whereas 7 μM NO-PAW could induce 18% apoptotic effect. In addition, we assumed that both DBD plasma and NO-PAW could induce HeLa cell apoptosis by facilitating an accumulation of intracellular reactive oxygen and nitrogen species (RONS). Although further detail on the molecular signal pathway is still needed, DBD and NO-PAW could become promising applications for effective and safe clinical trials for cancer therapy.

Non-thermal biocompatible plasma (NBP) has recently emerged as an attractive tool for surface modification of biomaterials in tissue engineering. Three dimensional chitosan scaffolds have been widely used in bone tissue engineering due to biodegradable and biocompatible properties. The present study aimed to evaluate osteogenic potential of NBP treated chitosan scaffold. The surface characteristics of scaffolds were analyzed by scanning electron microscopy (SEM) and X-ray diffraction (XRD), cell proliferation and differentiation was tested with osteoprogenitor cell line MC3T3-E1. The results show that NBP modified scaffold increase cell metabolic by MTT assay and live/dead assay. More importantly, we evidenced enhancement of osteogenic differentiation on NBP treated scaffolds by an increase of alkaline phosphatase (ALP) activity, high degree of extracellular mineralization and up-regulated osteogenic marker genes expression level. The findings in our study highlighted NBP as the innovative method to modified chitosan scaffold and to fine-tuning the scaffold a more suitable and beneficial biomaterial for in vivo bone tissue engineering and clinical bone defects therapies.

Recently, a growing body of evidence has shown the role of reactive species as secondary messengers in cell proliferation and differentiation, as opposed to the harmful metabolism byproducts that they were previously solely recognized as. Thus, the balance of intracellular reduction-oxidation (redox) homeostasis plays a vital role in the regulation of stem cell self-renewal and differentiation. Nonthermal biocompatible plasma (NBP) has emerged as a novel tool in biomedical applications. Recently, NBP has also emerged as a powerful tool in the tissue engineering field for the surface modification of biomaterial and the promotion of stem cell differentiation by the regulation of intracellular redox biology. NBP can generate various kinds of reactive oxygen species (ROS) and reactive nitrogen species (RNS), which may play the role of the second passenger in the cell signaling network and active antioxidant system in cells. Herein, we review the current knowledge on mechanisms by which NBP regulates cell proliferation and differentiation through redox modification. Considering the importance of redox homeostasis in the regulation of stem cell differentiation, understanding the underlying molecular mechanisms involved will provide important new insights into NBP-induced stem cell differentiation for tissue engineering.

Low‐temperature plasma, an engineered technology to generate various reactive species, is actively studied in cancer treatment in recent years, yet mainly by using a traditional 2D cell culture system. In this study, we explored the effect of the plasma‐activated medium (PAM) on lung cancer cells in vitro and in vivo by using a 3D cell culture model. The results showed that PAM markedly inhibited 3D spheroid formation and downregulated stemness‐related gene expression. We found that reactive oxygen species (ROS) penetrated throughout the whole spheroids and induced cell death surrounding and in the core of the tumor spheroid. Besides, PAM treatment suppressed migration and invasion of lung cancer cells and downregulated epithelial‐mesenchymal transition‐ (EMT‐) related gene expression. In the mouse xenograft model, the tumor volume was significantly smaller in the PAM‐treated group compared with the control group. By using transcriptome sequencing, we found that PI3K/Akt and MAPK pathways were involved in the inhibition effects of PAM on lung cancer cells. Therefore, our results indicated that PAM exhibits potential anticancer effects on lung cancer and provides insight into further exploration of PAM‐induced cell death and translational preclinical use.

The application of non-thermal bio-compatible plasma (NBP) for stem cell differentiation is promising in tissue engineering. However, the differentiation efficiency of NBP treatment on various types of human tissue-derived stem cells and the underlying mechanisms is yet understood. This study is the first time to investigate the role of NBP in inducing differentiation and its potential molecular mechanism by using human bone marrow-derived stem cells (hBMSCs) and human periodontal ligament-derived stem cells (hPDLSCs). Our results showed that NBP promote osteogenic differentiation of hPDLSCs more effectively than hBMSCs under the same treatment condition, indicating a tissue-dependent manner of NBP interacts with stem cells. Furthermore, an increase of intracellular reactive oxygen species (ROS) production of hPDLSCs and antioxidant enzymes activation was observed after NBP treatment. Particularly, mitogen-activated protein kinases (MAPKs) level was also increased and in consistent with ROS level increase. Taken together, this study revealed that with NBP induction, hPDLSCs is a more suitable stem cell source than hBMSCs for bone regeneration and tissue engineering, and ROS-induced activation of MAPKs are possibly involved in the osteogenic differentiation process.