## 1 Introduction  
The extent rate of melanoma is rising consistently and faster than any other malignancy over the last few years worldwide [1]. In South Korea only, the incidence rate of cutaneous melanoma in women increased continuously every year due to poor prognosis at advanced stages of the disease [2]. Patients diagnosed with melanoma in its later stages have limited treatment possibilities, and intermediate survival ranges from 6 to 12 months in clinical trials [3]. Despite the restricted treatment options over the past few years, some molecular targeted and immune therapies are in regular practice to diagnose and treat melanoma [4; 5; 6]. Presently, the US Food and Drug Administration (FDA) has approved a combination of drugs to treat advanced-stage melanoma. Most of them contain Interleukin-2 and dacarbazine in different combinations, but a very low percentage of patients respond to them [3]. Some researchers recently discovered and claimed that the mixture of chemical compounds showedpromising potential activity against malignant melanoma, like veumarafenib (zelboraf), dabrafenib (Tafinlar), and encorafenib (Braftovi), that primarily interact with the BRAF gene and inhibit its activity [7; 8; 9]. Nowadays, standard therapies for actual melanoma treatment are using certain immunotherapies. Apart from BRAF, immunosuppression of the antitumor immune response by certain immune regulatory molecules and processes, such as T-lymphocyte-associated antigen 4 (CTLA4) and programmed cell death 1 (PD-1), is of great interest [10]. In the past few years, cold atmospheric plasma (CAP) has emerged as an effective tool for treatment against several kinds of cancers [11]. The most important aspect of anticancer therapy is the selective eradication of cancer cells without influencing healthy cells [12]. Cold plasma plays a pivotal role in maintaining this cellular homeostasis [13]. CAP acts as a plasma medicine in the biological system because of the production of partially ionized clouds of ions, electrons, and reactive neutral species, like reactive oxygen species (ROS), hydroxyl radicals (OH\(\bullet\)), and nitrogen dioxide (NO\({}_{2}\)), by its direct application [14]. Further studies revealed by some researchers concluded that cancer cells are more sensitive to its destructive effect than normal cells, which help CAP as a promising application in cancer therapy [15; 16; 17]. Another aspect of plasma medicine depends on its indirect application (off-site treatment) in which CAP is applied to some liquid medium, which leads to an increase in the reactive species within the liquid, and hence, helps in the reduction of cancer and other dreadful diseases [16]. After the CAP treatment, the plasma-treated liquid is possessed with numerous long-lived RONS, which can be retained from hours to several days, and hence, can act as a replaceable agent of CAP where direct CAP treatment is not possible [18]. Plasma-Activated Media (PAM) generates some short-lived reactive species like OH radicals, which induced apoptosis in both extracellular (culture medium) and intracellular [19]. These short-lived species lead to the development of long-lived species, like H\({}_{2}\)O\({}_{2}\), which acts as an effective apoptosis inducer and can be applied to treat cancer cells [20; 21]. In recent times, new and advanced techniques have been used in the management of melanoma reduction to understand the disease behavior and help in the development and testing of novel therapeutic strategies. As a different approach to treating melanoma, our previous publication showed promising results of the use of CAP and PAM when used with silymarin nanoemulsion in _in vitro_ and _in vivo_ studies [22]. For the biological aspects of understanding melanoma, animal models contributed greatly as a useful tool for testing possible treatment approaches.