Purpose: This research was conducted to analyse and compare a Renuvion hand piece and TheraDep's deposition device at various gas flow rates and power settings. Both devices were powered by an Apyx Ultimate Electrosurgical generator. Methods: Current, voltage, optical emission spectroscopy (OES) and thermal imaging were used to investigate the resulting plasma intensities and species being ionised. It was found that the Renuvion device could deliver 5.6 to 8.2 W of power whereas the Deposition Accessory was significantly less at between 1.9 and 4.2 W depending on the power setting selected. Differences in the OES results were also observed which are thought to have resulted from variations in the atmospheric entrainment between the two devices. Results: Taken in combination, the electrical, optical emission and thermal data analysis has demonstrated that the addition of the Deposition Accessory transforms the Renuvion electrosurgical tool from a device that operates in the coagulation and cutting arena into a device with a dramatic reduction in the energy delivered to the target tissue. Consequently, this limits the localised heating of adjacent tissue and results in a device that has controlled coagulation capability and one which is not capable of cutting tissue. Conclusions: This offers significant potential to allow clinicians to evaluate applications in plasma medicine using an FDA cleared electrosurgical power source.
Preventing nosocomial infections is one of the most significant challenges in modern medicine. The disinfection of medical facilities and medical devices is crucial in order to prevent the uncontrolled spread of bacteria and viruses. Cost-effective, eco-friendly and fast-acting antibacterial coatings are being developed as the prevention of bacteria and viruses' multiplication on various surfaces. One of the possibilities to create such antimicrobial coatings can rely on a photoactive material, that produces singlet oxygen. However, a remote production of the singlet oxygen and disinfection of the desired surface is a time-consuming process. Hence, a coating material that would autonomously produce singlet oxygen employing ambient light will have a significant impact on the shortening of the disinfection time; leading into an increased number of patients that can be cured in one facility. In this work, an ultra-fast and eco-friendly method for decreasing the disinfection time of the photoactive surface is presented. The atmospheric pressure plasma surface treatment on the hydrophobic carbon quantum dotspolydimethylsiloxane nanocomposite is employed. The plasma-treated samples exhibited improved antibacterial properties compared to non-plasma treated samples, with the best results obtained after only 30 seconds of plasma treatment. The short duration and the scalability potential of the here described method open new possibilities of how to improve the already existing antibacterial coatings.
Breast cancer is a heterogenous disease which can be classified into subtypes by the presence or absence of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor 2 receptor (HER2). Cold atmospheric plasma (CAP) has been shown to be a potential treatment for cancer. In this report, a 92-99% reduction of viability by CAP treatment was achieved across all seven tested breast cancer cell lines (p <= 0.05). Increasing treatment duration and power significantly reduced breast cancer cell viability (** f(()(2,)(2)) <= 0.0176, ***f ((5,14)) <= 0.0033). The authors are the first to report that breast cancer sensitivity to CAP is based on receptor status. Cells with identical receptor status showed the least difference in CAP sensitivity (p <= 0.05), the difference being 33% between the two ER +/PR + /HER2- cell lines (p <= 0.05) and 22-44% between the three TNBC cell lines (p <= 0.05). HER2-negative cell lines, irrespective of ER/PR status, also showed <= 50% difference in CAP sensitivity (p <= 0.05). Moreover, demonstration of ER-/PR-/HER2 + CAP susceptibility and ER + /PR + /HER2 + CAP resistance suggests that ER/PR status is a significant factor in determining CAP sensitivity in HER2-positive cells. Our novel findings on CAP sensitivity will provide insight on how to optimize CAP treatment to better overcome CAP resistance and thus prevent tumor recurrence.
Background: Nitric oxide (NO) is a gaseous molecule produced by NO-synthases that regulates a wide range of physiological and pathophysiological processes. Endogenous NO signaling system plays an important role in all stages of wound healing. In 1998, a team from Bauman Moscow State Technical University and Sechenov First Moscow State Medical University (Sechenov University) developed the Plason device. Plason generates NO-containing air-plasma flow from atmospheric air and allows to apply required doses of NO to affected areas. It was shown, that exogenous NO penetrates through a wound surface, an intact skin, mucous membranes and a cornea, which leads to the wide range of applications. The experimental and clinical studies of 1992-2006 proved exogenous NO-therapy to be beneficial in patients with extensive purulent wounds, trophic ulcers, diabetic foot syndrome, purulent-inflammatory disease of soft tissues and scars. Methods: Up to now Plason is the only certified in Russia and Europe medical device generating NO from atmospheric air at high therapeutically significant concentrations (500-1200 ppm). It is used in Russia, Ukraine, Belarus and Slovakia. Other air-plasma devices, "Tianox" (Sarov, Russia) and "iNO pulse" (Bellerophon Inc., USA) generated low concentrations of NO (10-100 ppm) were found effective against severe acute respiratory syndrome and recently tested for treatment of COVID-19. The purpose of this review is to acquaint global medical community with the extensive experience of Russian clinicians in NO-therapy. It covers the NO-therapy studies in multiple medical fields published after 2006. Results and conclusions: Application of the Plason device was beneficial in treatment of wounds, purulent, traumatic and scar lesions, joint diseases, including sports injuries, as well as in abdominal surgery, dentistry, ophthalmology, otolaryngology and other areas of medicine.
Background: To optimize wound healing, the use of low-temperature atmospheric pressure plasma (ionized gas) has been proposed as an innovative therapeutic method for treating extensive and chronic wound. The aim of this study was to evaluate the specific effects of an indirect helium plasma treatment in a sheep surgical wound model based on clinical, histopathological and molecular analyses. This study was part of a larger study aimed at assessing five different wound healing improvement methods. Methods: Six sheep were used in this study. Six square wounds were performed on the back of each sheep. Five of the lesions were used to analyze the effect of five different treatments, one of which was the plasma treatment. The sixth lesions was used as control. A 2-min plasma treatment was daily performed, using a radiofrequency helium plasma source for indirect treatments, until complete wound healing was obtained. Biopsies were performed at two and six weeks. The plasma effects were tested through clinical evaluation, histopathological and immunohistological evaluation, real time PCR analysis, bacteriological evaluation and intracellular ROS evaluation. Results: Wound closure time for the plasma-treated wounds was quicker than that of the control group. Plasma treatment drastically reduced the bacterial load in the wounds. A strong increase in intracellular ROS was observed. After six weeks, the inflammatory process was significantly reduced in the plasma-treated case with respect to the control. The plasma treatment was found to lead to an anticipated induction of blood vessel formation, as detected through mRNA expression of VEGF. The high Ki67-positivity at two weeks indicated a strong stimulation of cell proliferation induced by the plasma treatment. The expression of the hair keratine (hKER) at six weeks indicated that the plasma was promoting hair regrowth. The plasma treatment induced an increased rate of reformation of cutaneous adnexa at six weeks. Conclusions: The obtained results suggest that the plasma action, at the used dose, induces an increase in cell proliferation, a reduction of inflammation, a reduction of the bacterial load, a stimulation of blood vessel formation, and an improvement in the formation of cutaneous adnexa with a positive consequence on hair regrowth.
Background: Cold atmospheric plasma (CAP) treatment has shown beneficial effects on wound healing treatment. Methods: A patient had a lesion on the middle finger of the left hand that was indicated to skin flap grafting. The patient then received the CAP treatment using a plasma device with a dose of 30 s/cm(2)/day. The patient's ulcer was treated according to standard procedure, and the ulcer's size, infection status, and sensation at the irradiation area were recorded each week. Results: After five weeks of this CAP treatment, the ulcer was completely epithelialized. Infection status and sensory abnormalities were not detected in the area of irradiation. Conclusion: CAP treatment may be a viable choice to treat patients wherein skin-grafting has failed and where the lesions have tendon and bone exposure.
Background: Acne vulgaris is a multifactorial skin disease that may be triggered by the presence of the bacteria Cutibacterium acnes and is characterized by an increase in the sebum excretion. Cold atmospheric plasma (CAP) represents an innovative technological device that uses plasma, the fourth state of matter, to treat several skin conditions, considering its bactericidal potential and its efficacy in promoting tissue proliferation. Objectives: In the light of its well-known antibacterial properties and its promising regenerative properties, we assumed that CAP could be highly effective to treat acne vulgaris both in its acute phase and in the long-term cicatricial period. Methods: We reported the cases of two young patients affected by mild-moderate acne vulgaris of the face treated with CAP. In addition to the evaluation of inflammatory skin lesions, treatment efficacy was assessed by sebometry and trans epidermal water loss (TEWL) evaluation. Results: In both patients, acne lesions reduced after treatment. Both sebum excretion parameters and TEWL improved considering basal values. Conclusion: Despite the small number of patients enrolled, our data support CAP effectiveness in patients with mild to moderate acne vulgaris poorly responsive to other conventional treatments.
The aim of this study was to evaluate color change, mineral composition and topography of the enamel after the pre-treatment with non-thermal atmospheric plasma (NTAP) followed by 38% hydrogen peroxide (HP) bleaching. Buccal enamel of bovine incisors teeth was stained with black tea solution and then divided into five groups (n = 12), according to the plasma application time: PL1 (one minute) or PL2 (2 min) and number of HP applications: HP1 (one) and HP3 (three). The following groups were investigated: HP3, PL1 + HP1, PL1 +HP3, PL2 + HP1 and PL2 + HP3. Four color measurements: initial (untreated), after staining and after bleaching (first and second sessions), were performed using an intraoral spectrophotometer (Vita Easyshade). Energy Dispersive X-ray Spectroscopy (EDX) was used to analyze the mineral composition of the enamel before and after two bleaching sessions. Laser confocal microscopy was used to analyze the enamel topography. The pre-treatment with NTAP did not improve the whitening results. The use of NTAP preserves the concentration of calcium and phosphorus, only HP3 group experimented a significant decrease in the mineral concentration. All the bleached groups, with or without NTAP treatment increased their enamel roughness. The application of NTAP did not influence the color change, but it was important to maintain the mineral composition of bleached teeth. All the groups increased their roughness level.
Background: Cold atmospheric plasma (CAP) has antimicrobial properties and may have a role in decontamination. However, CAP can coagulate blood, which could lead to the fixing of blood or proteins and make surface decontamination challenging after blood spillages. Methods: In vitro assays were performed to assess whether CAP treatment of a surface impaired removal of a blood soil from steel through visual inspection and bicinchoninic acid assays. To assess if remaining blood proteins promoted biofilm formation, crystal violet biomass analysis was performed. Results: In an in vitro study, CAP treatment did not hinder the removal of blood and pre-treatment decreased protein remaining on the surface. CAP inhibited Staphylococcus aureus biofilm formation on blood-soiled surfaces. Conclusions: CAP does not enhance the persistence of blood and protein on steel surfaces. Biofilm formation after CAP treatment of blood soiled steel was decreased possibly due to changes in electrostatic charge on surfaces. CAP should be further evaluated as an alternative surface decontaminant.
Background: Cold atmospheric plasma (CAP) is widely used in the cancer therapy field. This type of plasma is very close to room temperature. This paper illustrates the effects of CAP on breast cancer tissues both in vivo and in vitro. Methods: The mouse mammary adenocarcinoma cell line AN3 was used for the in vivo study, and the MCF7, AMJ13, AMN3, and HBL cell lines were used for the in vitro study. A floating electrode-dielectric barrier discharge (FE-DBD) system was used. The cold plasma produced by the device was tested against breast cancer cells. Results: The induced cytotoxicity percentages were 61.7%, 68% and 58.07% for the MCF7, AMN3, and AMJ13 cell lines, respectively, whereas the normal breast tissue HBL cell line exhibited very little or no cytotoxicity. Reactive oxygen species (ROS) were measured, and we found that more ROS were generated under the impact of CAP in cancer cells, whereas the normal HBL cell line had the lowest ROS level. The in vivo study showed that CAP treatment could reduce the volume of treated tumors compared to those in untreated mice. Conclusions: CAP has anticancer effects both in vitro and in vivo and this effect is mediated by the ROS and induce apoptosis in p53 independent pathway. the current method is promising for breast cancer therapy.
Background: It is known that exogenous nitric oxide (NO) has a bactericidal effect, activates proliferation of fibroblasts, enhances angiogenesis and collagen synthesis, accelerates maturation of granulation tissue. Objective: The purpose of this research was to study the dose-dependent therapeutic effect of nitric oxide (NO) in NO-containing gas flows (NO-CGF) applied to the wound surfaces. Methods: The experiment was performed in 36 white Wistar rats with full-thickness skin wounds with an area size of 3 cm(2). We used Plason device modified with an experimental plasma-chemical generator of nitric oxide (manipulator) for the following parameters of NO-CGF in the wound surface area: mass flow rate of nitric oxide 2.0 mg/s, axial gas flow velocity 9 m/s, nitric oxide content 1340 ppm, nitrogen dioxide content 108 ppm, gas flow temperature 39 degrees C. Each group included 6 animals. The mass of nitric oxide delivered to the wound varied by the exposure time of 15, 30, 60, 120 and 360 seconds and was 30, 60, 120, 240 and 720 mg, respectively. Wounds were left untreated in the control group. Wound tissues were excised 4 days after the operation and studied with histological, semi-quantitative and morphometric methods. Results: Histological analysis revealed beneficial effects of NO therapy on wound healing, including reduction of microcirculatory disorders and exudate volume and acceleration of fibroblast proliferation and collagen synthesis. We proved that the most effective stimulation of the wound healing process occurred with a duration of exposure of 120 and 360 seconds (integral mass doses of NO on the wound surfaces were 240 mg and 720 mg, relative mass doses of NO per wound areas were 80 mg/cm(2) and 240 mg/cm(2), respectively). We did not identify the overdose phenomena after prolonged exposure of the wound to NO. Conclusion: Our results are important for further improvement of the technique and the standardization of NO-therapy for the treatment of wounds, inflammatory and destructive processes in clinical practice.
Purpose: In Endodontics, in order to complete a root canal treatment, the use of guttapercha to completely seal the root apex must be coupled with endodontic cements known as sealers to improve the adhesion with dentine, despite their cytotoxicity and solubility. The present study investigates the effect of enhancement of adhesion between these materials and Cold Atmospheric Plasma (CAP)-treated dentine of the apical region of ex-vivo teeth as a first step towards the reduction on the use of cytotoxic materials and the use of CAP in-vivo at a clinical stage. Methods: A dielectric barrier discharge (DBD) helium plasma jet was used to treat for 180 sec the shaped root canal dentin. Pushout tests and confocal microscopy analysis have been performed to evaluate the effect of cold plasma treatment in terms of adhesion performances and interaction between filling materials and dentin respectively. Results: The pushout WA results highlight how a 180 seconds CAP treatment of dentin promotes an enhancement of bonding strength between filling materials and dentine in all investigated cases. In particular, when the dentine is CAP-treated in the apical region, the adhesion performances of guttapercha achieved a relevant increase (similar to + 200%), revealing how it was able to "self-bond" with the substrate with results comparable with the conventional procedure but without any sealer application, while the conventional (guttapercha + sealer) apical sealing procedure shows an improvement around + 50%. Moreover, confocal images qualitatively confirmed the higher spreading and penetration of both guttapercha and sealer into dentinal tubules. The ergonomics of the prototype has been also improved to comply with the dimensions of devices commonly used in the dental field, with a plasma source diameter of 21.5 mm and with a choice of materials to minimise its weight. Conclusion: CAP treatment of dentin increases the adhesion performances and paves the way for a sealer-free and safer procedure for apical restoration.
Background: In recent years, cold physical plasma gained significant interest as a new anticancer tool. Cold plasmas are versatile apparatuses in medicine and material processing. Among different cold plasma devices, the argon plasma jet is an effective apparatus and favorite for researches in plasma medicine science. Methods: We developed an argon plasma jet for cancer treatments. Using optical emission spectroscopy, the newly developed plasma jet produced a substantial amount of hydroxyl radicals and other reactive oxygen and nitrogen species known to show cytotoxic effects at higher concentrations. In this work, we studied the efficacy of the argon-based plasma jet treatment in murine B16 melanoma cells in vitro and in vivo. In vitro, cell viability, apoptotic gene expression, and cell death patterns were assessed at 24 h and compared against non-malignant murine dermal fibroblasts (MDF). In vivo, B16 cells inoculated into the flank of the mice developed melanoma tumors, which were plasma-treated for 5 min on days 1, 5, and 10 or were left untreated. Results: Melanoma cells were significantly more sensitive to plasma treatment than MDF under the same in vitro conditions (p = 0.00023). The results revealed that the argon-based plasma jet damaged B16 melanoma cells in vitro selectively without a detrimental effect on non-malignant MDF cells. In the animal model, the tumor volume was significantly reduced after three treatments with the plasma jet(p < 0.01). Conclusions: The argon plasma jet showed therapeutic anti-melanoma efficacy in vitro and in vivo, which may spur new research lines in melanoma therapy.
Background: Low temperature plasma is an ionised gas generated at atmospheric pressure with the temperature of heavy particles (ions, molecules, atoms) close to room temperature. Recent progress highlighted the efficiency of plasma to induce cell death in many cancer cell lines. In the current context of the emergence of the use of cold plasma for medical purposes, this article aims to study the effect of a cold plasma of helium on ex-vivo prostate cells. Methods: A cold atmospheric plasma torch was developed to treat different tumor cells. Parameters like electric voltage, gas flow, cell/torch distance, and exposure time were studied to improve the effects of plasma onto two human prostate tumor cells lines, with different metastatic potential, LNCaP and PC3 and then, compared to human prostate epithelial cells P69. The plasma torch operated by a voltage up to 10 kV, without any risk of arcing, exhibited a visible plasma plume up to about 70 mm, with a temperature close to room temperature. The induced temperature rise due to the plasma treatment is measured to ensure that observed results are not thermal effects. After plasma treatment, microscopy and MTT analyses are achieved to measure cells viability. An ordinary one-way ANOVA test were used between two groups with p-value less than 0.05. Results: An optimized protocol leads to the tumor cells death with regard to cell viability. Cold plasma treatment exhibits differential effect, regarding tumor status of cells without any significant influence of thermal effect (temperature increases of only few degrees Celcius above room temperature). The results show that non-tumor cells P69 are less sensitive to cold plasma exposure with a cell viability of 67% after 120 s of treatment, compared to tumor cell lines LNCaP and PC3, respectively at cell viability of 29% and 23% for the same treatment time. Conclusion: The cold helium plasma treatment showed a selective effect on prostate cells, with an enhanced apoptosis way for tumor cells, whereas the healthy cells are more resistant to the treatment. Moreover, during the treatment, the surface temperature rises only few degrees Celcius. This result is very promising for future in-vivo treatments.
Cold plasma sterilization offers an efficient way to sterilize medical components and instruments. This paper reports using a magnetized plasma to realize low-temperature sterilization. A radio frequency dielectric barrier discharge is created in a quartz tube using a mixture of argon and oxygen gas. A uniform amount of Escherichia coli is applied onto glass slides and exposed to the plasma afterglow at different pressures with and without a magnetic field. Optical emission spectroscopy is used to identify the plasma species present. The magnetic field significantly promotes the intensity of the plasma and the sterilization efficiency. A process gas pressure of 100 mTorr presents the most effective treatment with a sterilization time less than one minute and sample temperature below 32 °C.
Purpose: To evaluate the effect of application of 3% air in helium cold atmospheric plasma jet, using an inexpensive device termed iCAP, in corneal scratch wound closure in vitro and the treatment of Pseudomonas aeruginosa (P. aeruginosa) keratitis in vivo. Methods: Thermal imaging to measure temperature of surfaces to which iCAP was applied and UV energy density delivered by iCAP were measured. Scratch wounds inflicted on in vitro cultures of a human corneal epithelial cell line were treated with iCAP and wound widths at various times post-application were measured. Rabbit eyes infected with P. aeruginosa were treated with iCAP and slit lamp biomicroscope examination conducted to determine corneal health outcomes 25 h post infection. Corneal homogenates were plated on agar and viable bacterial colonies enumerated to determine the effect of iCAP on bacterial load in vivo in P. aeruginosa keratitis. Results: iCAP was shown to operate in the non-thermal regime and also shown to deliver much lower UV energy density than that necessary to cause harmful effects on ocular tissue. iCAP treatment significantly improved the rate of scratch wound gap closure in vitro in a human corneal epithelial cell line compared to controls. In vivo, iCAP treatment of P. aeruginosa keratitis infection in the rabbit eyes (N = 20) significantly reduced the incidence of corneal ulcer (P = 0.003) and corneal edema (P = 0.011) and significantly improved total cornea health (P = 0.034) compared to untreated (N = 10). Finally, in vivo iCAP treatment of P. aeruginosa keratitis infection in the rabbit eyes (N = 19) significantly reduced bacterial loads (P = 0.012) compared to untreated (N = 9). Conclusion: Our results strongly suggest that iCAP treatment was effective in improving corneal epithelial defect closure in vitro, reducing ulcer formation and decreasing inflammation in P. aeruginosa infected corneas in vivo and decreasing bacterial loads in P. aeruginosa infected corneas in vivo which led to improved overall cornea health outcomes in vivo. Further studies to investigate iCAP's safety and efficacy against other infectious microbes responsible for causing ulcerative keratitis, with and without co-treatment with antimicrobial therapies are warranted.
Purpose: The possibility of obtaining plasmas at low temperatures and atmospheric conditions, the Cold Atmospheric Plasmas (CAPs), has triggered several studies on its possible medical applications. The use of CAPs devices applied to oncological treatments is still in a pre-clinical stage, but early investigations have shown promising results, both in vitro and in vivo. In this work, is reported the study of a custom made Cold Atmospheric Pressure Plasma jet device for indirect treatment of Squamous Cell Carcinoma. Methods: The plasma plume produced by the jet device was characterized by optical emission spectroscopy and the vulnerability of two different cell lines, SCC-15 (Squamous Cell Carcinoma) and HGF-1 (non-cancer cell line) to different working parameters factors such as the volume of medium used on indirect plasma treatments, the number of cells used in the biological assays and the volume of treated medium in contact with the cultured cells were systematically tested and studied. Results: The effectiveness of CAPs' treatments is time, volume and cell dependent. Cancer cells, specifically SCC-15 cells have shown to be more sensitive to indirect plasma treatments than the non-cancer cell line used in the present work. Conclusions: According to the obtained results, this study showed that thein vitro anti-cancer capacity of the developed jet plasma device will be higher when small volumes of medium are used together with moderate plasma treatment times, keeping the non-cancerous cells almost unaffected.
Cancer continues to be a significant threat to human health. Oncotherapy, therefore, relies on the combination of different approaches to increase a patient's chances. If therapeutic options are exhausted, effective palliation can at least still improve the quality of life. Over two decades ago, electrochemotherapy (ECT), which is based on pulsed electric field (PEF) exposures, was introduced in palliative care to alleviate the burden imposed by tumors, such as malignant melanoma and breast cancer. In this case, pulsed electric fields with a duration in the range of microseconds permeabilize cell membranes and permit reducing dosages of cytotoxic drugs and accordingly of associated systemic side effects. More recently, exposures to pulsed electric fields, shorter than the cellular plasma membrane charging time, have been found to affect subcellular structures and cell functions directly, i.e., without additional drugs. Instead of cytotoxic drugs, also the delivery of genes and calcium is currently investigated for alternative treatment options. Another way to induce tumor cell death is the introduction and/or generation of reactive species in the cellular environment by the application of cold physical plasma, resulting in the activation of redox signaling pathways. These latest developments encourage considering new treatment options. Of particular interest might be the possibility to promote the uptake of plasma-generated species by the combination with PEF-exposures. Hence, we review the latest developments in clinical and experimental ECT and PEF research in oncology and rationalize a combined treatment with plasmas for palliative therapy in patients.
Purpose: Venous, neuropathic, and mixed (venous and arterial) ulcers are complex chronic wounds that have a tremendous long-term impact on the mortality, morbidity, and quality of life of patients. In this work it was evaluated the efficacy of the non-thermal helium plasma at atmospheric pressure applied in patients with this suffering. Methods: Using three elements: a RF generator, a needle-type reactor, and helium gas flow, for non-thermal plasma generation were used. The power density on the tissue of patients was of 0.6-0.67 W/cm(2). The application time was 30 s/cm(2) daily, at 5 to 10 mm of distance from the wound. Results: The thirty-two patients with venous and mixed ulcers indicated that they had a decrease progressively of the pain with the therapy, without the need to continue with analgesics. Besides, the ten patients with chronic wounds showed 100% healing in neuropathic ulcers. For eight patients with venous leg ulcers, the percentage was reduced to 75% healing. While for fourteen patients with mixed leg ulcers, the rate of healing was 59%. Conclusion: The non-thermal plasma generated in a needle reactor with helium gas is a promising candidate for clinical and therapeutic use in the treatment of neuropathic leg ulcers, with time reduction healing, and it can be applied everyday. In the study conducted, it was found that the non-thermal plasma applied to patients with autoimmune diseases clinically was not beneficial.
