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Cold atmospheric plasmas (CAPs) within recent years have shown great promise in the field of plasma medicine, encompassing a variety of treatments from wound healing to the treatment of cancerous tumors. For each subsequent treatment, a different application of CAPs has been postulated and attempted to best treat the target for the most effective results. These treatments have varied through the implementation of control parameters such as applied settings, electrode geometries, gas flow, and the duration of the treatment. However, with such an extensive number of variables to consider, scientists and engineers have sought a means to accurately control CAPs for the best-desired effects in medical applications. This paper seeks to investigate and characterize the historical precedent for the use of plasma control mechanisms within the field of plasma medicine. Current control strategies, plasma parameters, and control schemes will be extrapolated through recent developments and successes to gain better insight into the future of the field and the challenges that are still present in the overall implementation of such devices. Proposed approaches, such as data-driven machine learning, and the use of closed-loop feedback controls, will be showcased as the next steps toward application.

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BACKGROUND: Cold atmospheric plasma (CAP) therapy has been recognized as effective treatment option for reducing bacterial load in chronic wounds, such as adult ventricular assist device (VAD) driveline exit-site infections. Currently, there have been no reports on the safety and efficacy of CAP therapy for pediatric cannula infections and inflammations in paracorporeal pulsatile VADs. METHODS: The mechanical strength of Berlin Heart EXCOR cannulas were tested both before and after CAP treatment (SteriPlas, Adtec Healthcare Limited, UK) to prove material safety. A ring tensile test of 20 untreated and 20 CAP-treated (5 min) EXCOR cannulas (Ø12mm), assessed the force at the breaking point of the cannulas (Fmax), at 25% (F25%) and 50% (F50%) of the maximum displacement. Additionally, the scanning electron microscope (SEM) micrographs for both groups examined any surface changes. Finally, the case of a 13-year-old male EXCOR patient with cannula infections, treated with CAP over 100 days, is presented. RESULTS: The in vitro measurements revealed no statistically significant differences in mechanical strength between the control and CAP group for F25% (8.18 ± 0.36 N, vs. 8.02 ± 0.43 N, p = 0.21), F50% (16.87 ± 1.07 N vs. 16.38 ± 1.32 N, p = 0.21), and FMAX (44.55 ± 3.24 N vs. 42.83 ± 4.32 N, p = 0.16). No surface structure alterations were identified in the SEM micrographs. The patient's cannula exit-sites showed a visible improvement in DESTINE wound staging, reduction in bacterial load and inflammatory parameters after CAP treatment without any side effects. CONCLUSION: Overall, CAP therapy proved to be a safe and effective for treating EXCOR cannula exit-site wound healing disorders in one pediatric patient, but further studies should investigate this therapy in more detail.

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The study aimed to explore the antimicrobial efficacy of grape seed extract (GSE) and cold atmospheric plasma (CAP) individually or in combination against L. monocytogenes and E. coli wild type (WT) and their isogenic mutants in environmental stress genes. More specifically, we examined the effects of 1% (wt/vol) GSE, 4 min of CAP treatment, and their combined effect on L. monocytogenes 10403S WT and its isogenic mutants ΔsigB, ΔgadD1, ΔgadD2, ΔgadD3, as well as E. coli K12 and its isogenic mutants ΔrpoS, ΔoxyR, and ΔdnaK. In addition, the sequence of the combined treatments was tested. A synergistic effect was achieved for all L. monocytogenes strains when exposure to GSE was followed by CAP treatment. However, the same effect was observed against E. coli strains, only for the reversed treatment sequence. Additionally, L. monocytogenes ΔsigB was more sensitive to the individual GSE and the combined GSE/CAP treatment, whereas ΔgadD2 was more sensitive to CAP, as compared to the rest of the mutants under study. Individual GSE exposure was unable to inhibit E. coli strains, and individual CAP treatment resulted in higher inactivation of E. coli in comparison to L. monocytogenes with the strain ΔrpoS appearing the most sensitive among all studied strains. Our findings provide a step toward a better understanding of the mechanisms playing a role in the tolerance/sensitivity of our model Gram-positive and Gram-negative bacteria toward GSE, CAP, and their combination. Therefore, our results contribute to the development of more effective and targeted antimicrobial strategies for sustainable decontamination.IMPORTANCEAlternative approaches to conventional sterilization are gaining interest from the food industry, driven by (i) the consumer demand for minimally processed products and (ii) the need for sustainable, environmentally friendly processing interventions. However, as such alternative approaches are milder than conventional heat sterilization, bacterial pathogens might not be entirely killed by them, which means that they could survive and grow, causing food contamination and health hazards. In this manuscript, we performed a systematic study of the impact of antimicrobials derived from fruit industry waste (grape seed extract) and cold atmospheric plasma on the inactivation/killing as well as the damage of bacterial pathogens and their genetically modified counterparts, for genes linked to the response to environmental stress. Our work provides insights into genes that could be responsible for the bacterial capability to resist/survive those novel treatments, therefore, contributing to the development of more effective and targeted antimicrobial strategies for sustainable decontamination.

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The miniaturization and widespread deployment of electronic devices across diverse environments have heightened their vulnerability to corrosion, particularly affecting copper traces within printed circuit boards (PCBs). Conventional protective methods, such as conformal coatings, face challenges including the necessity for a critical thickness to ensure effective barrier properties and the requirement for multiple steps of drying and curing to eliminate solvent entrapment within polymer coatings. This study investigates cold atmospheric plasma (CAP) as an innovative technique for directly depositing ultrathin silicon oxide (SiOx) coatings onto copper surfaces to enhance corrosion protection in PCBs. A systematic investigation was undertaken to examine how the scanning speed of the CAP deposition head impacts the film quality and corrosion resistance. The research aims to determine the optimal scanning speed of the CAP deposition head that achieves complete surface coverage while promoting effective cross-linking and minimizing unreacted precursor entrapment, resulting in superior electrical barrier and mechanical properties. The CAP coating process demonstrated the capability of depositing SiOx onto copper surfaces at various thicknesses ranging from 70 to 1110 nm through a single deposition process by simply adjusting the scanning speed of the plasma head (5-75 mm/s). Evaluation of material corrosion barrier characteristics revealed that scanning speeds of 45 mm/s of the plasma deposition head provided an effective coating thickness of 140 nm, exhibiting superior corrosion resistance (30-fold) compared to that of uncoated copper. As a proof of concept, the efficacy of CAP-deposited SiOx coatings was demonstrated by protecting an LED circuit in saltwater and by coating printed circuits for potential agricultural sensor applications. These CAP-deposited coatings offer performance comparable to or superior to traditional conformal polymeric coatings. This research presents CAP-deposited SiOx coatings as a promising approach for effective and scalable corrosion protection in miniaturized electronics.

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Context: Effective use of nonthermal atmospheric plasma (NTAP) to strengthen adhesive-dentin interfacial bonding while disinfecting with chlorhexidine (CHX). Aim: NTAP application at different time intervals on the dentinal shear bond strength (SBS) after pretreatment with 2% CHX as a cavity disinfectant. Settings and Design: The design of the study was an in vitro study. Materials and Methods: Forty permanent mandibular teeth were collected (n = 40) and the occlusal surfaces were flattened. For, all the specimens 37% phosphoric acid etching was done followed by pretreatment with 2% CHX as cavity disinfectant for 5 s. According to the surface treatment, divided into four groups of n = 10. Group I (Control): No NTAP pretreatment was done. Group II: NTAP pretreatment done for 15 s. Group III: NTAP pretreatment done for 30 s. Group IV: NTAP pretreatment done for 45 s. Later, all the specimens were treated with a bonding agent, incremental build-up of composite resin on the dentin surface was done and evaluation of SBS was done. Analyzed using One-way analysis of variance with a post hoc Tukey's test (P < 0.05). Results: Two percent CHX pretreatment as cavity disinfectant followed by NTAP application for 30 s (Group III) exhibited greater values compared to the control group (Group I). Conclusion: Two percent CHX pretreatment as cavity disinfectant followed by NTAP pretreatment for 30 s was found to exhibit better bond strength values compared to 15 s as well as 45 s.

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Cold atmospheric plasma (CAP) devices generate reactive oxygen and nitrogen species, have antimicrobial and antiviral properties, but also affect the molecular and cellular mechanisms of eukaryotic cells. The aim of this study is to investigate CAP treatment in the upper respiratory tract (URT) to reduce the incidence of ventilator-associated bacterial pneumonia (especially superinfections with multi-resistant pathogens) or viral infections (e.g., COVID-19). For this purpose, the surface-microdischarge-based plasma intensive care (PIC) device was developed by terraplasma medical GmbH. This study analyzes the safety aspects using in vitro assays and molecular characterization of human oral keratinocytes (hOK), human bronchial-tracheal epithelial cells (hBTE), and human lung fibroblasts (hLF). A 5 min CAP treatment with the PIC device at the "throat" and "subglottis" positions in the URT model did not show any significant differences from the untreated control (ctrl.) and the corresponding pressurized air (PA) treatment in terms of cell morphology, viability, apoptosis, DNA damage, and migration. However, pro-inflammatory cytokines (MCP-1, IL-6, and TNFα) were induced in hBTE and hOK cells and profibrotic molecules (collagen-I, FKBP10, and αSMA) in hLF at the mRNA level. The use of CAP in the oropharynx may make an important contribution to the recovery of intensive care patients. The results indicate that a 5 min CAP treatment in the URT with the PIC device does not cause any cell damage. The extent to which immune cell activation is induced and whether it has long-term effects on the organism need to be carefully examined in follow-up studies in vivo.

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This study proposes an affordable plasma device that utilizes a parallel-plate dielectric barrier discharge geometry with a metallic mesh electrode, featuring a straightforward 3D-printed design. Powered by a high-voltage supply adapted from a cosmetic plasma device, it operates on atmospheric air, eliminating the need for gas flux. Surface modification of polyethylene treated with this device was characterized and showed that the elemental composition after 15 min of plasma treatment decreased the amount of C to ~80 at% due to the insertion of O (~15 at%). Tested against Candida albicans and Staphylococcus aureus, the device achieved a reduction of over 99% in microbial load with exposure times ranging from 1 to 10 min. Simultaneously, the Vero cell viability remained consistently high, namely between 91% and 96% across exposure times. These results highlight this device's potential for the surface modification of materials and various infection-related applications, boasting affordability and facilitating effective antimicrobial interventions.

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Spinal cord injury (SCI) presents a critical medical challenge, marked by substantial neural damage and persistent functional deficits. This study investigates the therapeutic potential of cold atmospheric plasma (CAP) for SCI, utilizing a tailored dielectric barrier discharge (DBD) device to conduct comprehensive in vivo and in vitro analyses. The findings show that CAP treatment significantly improves functional recovery after SCI, reduces neuronal apoptosis, lowers inflammation, and increases axonal regeneration. These findings illustrate the efficacy of CAP in fostering a conducive environment for recovery by modulating inflammatory responses, enhancing neuronal survival, and encouraging regenerative processes. The underlying mechanism involves CAP's reactive oxygen species (ROS) reduction, followed by activating antioxidant enzymes. These findings position CAP as a pioneering approach for spinal cord injury (SCI) treatment, presenting opportunities for improved neural recovery and establishing a new paradigm in SCI therapy.

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Chronic wound infections are a silent pandemic in danger of becoming a global healthcare crisis. Innovations to control infections and improve healing are required. In the context of this challenge, researchers are exploiting plasma-activated hydrogel therapy (PAHT) for use either alone or in combination with other antimicrobial strategies. PAHT involves the cold atmospheric pressure plasma activation of hydrogels with reactive oxygen and nitrogen species to decontaminate infections and promote healing. This opinion article describes PAHT for wound treatment and provides an overview of current research and outstanding challenges in translating the technology for medical use. A 'blueprint' of an autonomous PAHT is presented in the final section that can move the management and treatment of wounds from the clinical setting to the community.

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Cold atmospheric plasma (CAP) is a promising alternative to antibiotics and chemical substances in dentistry that can reduce the risk of unwanted side effects and bacterial resistance. AmbiJet is a device that can ignite and deliver plasma directly to the site of action for maximum effectiveness. The aim of the study was to investigate its antimicrobial efficacy and the possible development of bacterial resistance. The antimicrobial effect of the plasma was tested under aerobic and anaerobic conditions on bacteria (five aerobic, three anaerobic (Gram +/-)) that are relevant in dentistry. The application times varied from 1 to 7 min. Possible bacterial resistance was evaluated by repeated plasma applications (10 times in 50 days). A possible increase in temperature was measured. Plasma effectively killed 106 seeded aerobic and anaerobic bacteria after an application time of 1 min per 10 mm2. Neither the development of resistance nor an increase in temperature above 40 °C was observed, so patient discomfort can be ruled out. The plasma treatment proved to be effective under anaerobic conditions, so the influence of ROS can be questioned. Our results show that AmbiJet efficiently eliminates pathogenic oral bacteria. Therefore, it can be advocated for clinical therapeutic use.

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The study examines the induction of apoptosis in colon cancer stem cells (CCSCs) within a 3D culture setting, employing an innovative cold atmospheric plasma (CAP) transmission method known as two-stage transferred cold atmospheric plasma (TS-TCAP). TS-TCAP is a partially or fully ionized non-thermal gaseous mixture that comprises photons, charged and neutral particles, and free radicals, which has gained traction in biomedical applications such as cancer therapy. TS-TCAP impacts CCSCs via a continuous, two-step transport process, facilitating the efficient delivery of reactive oxygen and nitrogen species (RONS). The key cellular factors of CCSCs impacted by TS-TCAP treatment, encompassing the secretion and expression levels of IL-6 and IL-8, apoptotic cell count, and expression of BAX, BCL-2, and KI-67 proteins, were evaluated using qrt-ELISA, Annexin V, and qrt-PCR procedures, respectively. The outcomes of CCSCs treatment with TS-TCAP reveal a notable rise in the number of apoptotic cells (P<0.0001), diminished secretion, and gene expression of IL-6 and IL-8 (P<0.0001), accompanied by favorable alterations in BCL-2 and BAX gene expression (P<0.0001). Additionally, a notable decrease in KI-67 expression was observed, correlating with a reduction in CCSCs proliferation (P<0.0001). As well, this study underscores the anti-cancer potential of TS-TCAP, showcasing its efficacy in reducing CCSCs survival rates. However, further pre-clinical and clinical trials are necessary to evaluate CAP's efficacy, safety, and potential synergistic effects with other therapies thoroughly. Overall, TS-TCAP presents a promising alternative for CCSCs treatment, pending further investigation and refinement.

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Cold atmospheric plasma (CAP) is a fledgling therapeutic technique for psoriasis treatment with noninvasiveness, but clinical adoption has been stifled by the insufficient production and delivery of plasma-generated reactive oxygen and nitrogen species (RONS). Herein, patches of air-discharge plasma-activated ice microneedles (PA-IMNs) loaded with multiple RONS are designed for local transdermal delivery to treat psoriasis as an alternative to direct CAP irradiation treatment. By mixing two RONS generated by the air-discharge plasma in the NOx mode and O3 mode, abundant high-valence RONS are produced and incorporated into PA-IMNs via complex gas-gas and gas-liquid reactions. The PA-IMNs abrogate keratinocyte overproliferation by inducing reactive oxygen species (ROS)-mediated loss of the mitochondrial membrane potential and apoptosis of keratinocytes. The in vivo transdermal treatment confirms that PA-IMNs produce significant anti-inflammatory and therapeutic actions for imiquimod (IMQ)-induced psoriasis-like dermatitis in mice by inhibiting the release of associated inflammatory factors while showing no evident systemic toxicity. Therefore, PA-IMNs have a large potential in transdermal delivery platforms as they overcome the limitations of using CAP directly in the clinical treatment of psoriasis.

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BACKGROUND: Cold atmospheric plasma (CAP) is a new therapeutic tool used to treat various skin diseases in humans and animals. OBJECTIVE: To evaluate the effect of CAP in the treatment of canine acute otitis externa (AOE). ANIMALS: Four client-owned golden retriever dogs with bilateral AOE. METHODS AND MATERIALS: After cleaning with a commercial ear cleanser, right ears (STANDARD group) were treated with an antibiotic/antifungal/corticosteroid combination and left ears (CAP group) were treated with CAP every three days for a total of four treatments. Cytological score and otitis index score (OTIS)3 were recorded for each ear on Day (D)0, D10 and D15. At D10 and D15, owners and investigators recorded an overall assessment. RESULTS: In both groups, OTIS3 and cytological score decreased over the study period. The overall assessment scale ranged from moderate to excellent in both groups. CONCLUSIONS AND CLINICAL RELEVANCE: Cold atmospheric plasma treatment showed equal therapeutic effect compared with a commercial topical anti-inflammatory and antimicrobial ear treatment.

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In this work, the effect of etching the surface of polymer matrix nanocomposites with atmospheric pressure plasma targeting to achieve enhanced dielectric properties was investigated. Polymer nanocomposites, with varying reinforcing phase content, were modified by atmospheric-pressure plasma resulting in an increase in the surface filler's concentration. Polymethyl methacrylate (PMMA) matrix nanocomposites reinforced with zinc oxide (ZnO) nanoparticles were prepared and dielectrically studied as a function of the nanoparticle content and the plasma modified surfaces. The electrical response of the composite systems was studied by means of Broadband Dielectric Spectroscopy (BDS) over a wide range of temperatures and frequencies. The dielectric permittivity increased with the embedded phase content and with plasma surface treatment. Energy density followed the same trend as dielectric permittivity, and the plasma-treated nanocomposite with the higher ZnO content exhibited approximately 27% higher energy density compared to the unreinforced matrix.

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Background: For reusable devices and device components, effective reprocessing is essential to prevent nosocomial infections. Aim: The objective of the study was to evaluate manual cleaning as the first step of reprocessing reusable glass probes of a device for generation of non-invasive physical plasma, in accordance with regulations. Methods: Two glass probes of the device were contaminated with human blood. For manual cleaning, both probes were cleaned with instrument cleaning agent and instrument brushes. Cleaning efficacy was evaluated by total protein measurement in the rinsing solution. Results: After manual cleaning of the two test glass probes, no protein from the test contamination with human blood could be detected. Neither the different design of the two probes nor the use of a hard or a soft instrument brush demonstrated any difference. Conclusion: Our data suggest that manual cleaning of glass probes achieves complete removal of organic contaminants. This should enable safe applications in clinical practice.

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In the present study, biopolymeric coatings of polyhydroxybutyrate (PHB) were deposited on 316L stainless steel substrates. The PHB coatings were developed using the spin coating method. To improve the adhesion of the PHB coating on the substrate, this method uses an atmospheric plasma treatment. Adhesion tests show a 156% increase in adhesion after 5 s of surface treatment. Raman spectroscopy analysis of the polymer shows the incorporation of functional groups and the formation of new hydrogen bonds, which can help us bind drugs and promote osteogenesis after plasma treatment. Additionally, the electrochemical behaviors in artificial body fluids (Hanks' solution) of the PHB coatings on the steel were evaluated with potentiodynamic tests, which revealed a decrease in the corrosion current and resistance to the transfer of the charge from the electrolyte to the 316L steel because of the PHB coating. All the PHB coatings were characterized using scanning electron microscopy and Raman spectroscopy after the electrochemical tests. This analysis confirmed the diffusion of electrolyte species toward the surface and the degradation of the polymer chain for the first 15 s of treatment with atmospheric plasma. These findings support the claim that plasma surface modification is a quick, environmentally friendly, and cost-effective method to enhance the performance of PHB coatings on 316L stainless steel for medical devices.

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Chondrosarcoma (CS) is a rare malignant bone sarcoma that primarily affects cartilage cells in the femur and pelvis. While most subtypes exhibit slow growth with a very good prognosis, some aggressive subtypes have a poorer overall survival. CS is known for its resistance to chemotherapy and radiotherapy, leaving surgery as the sole effective therapeutic option. Cold physical plasma (CPP) has been explored in vitro as a potential therapy, demonstrating positive anti-tumor effects on CS cells. This study investigated the synergistic effects of combining CPP with cytostatics on CS cells. The chemotherapeutic agents cisplatin, doxorubicin, and vincristine were applied to two CS cell lines (CAL-78 and SW1353). After determining their IC20 and IC50, they were combined with CPP in both cell lines to assess their impact on the cell proliferation, viability, metabolism, and apoptosis. This combined approach significantly reduced the cell proliferation and viability while increasing the apoptosis signals compared to cytostatic therapy alone. The combination of CPP and chemotherapeutic drugs shows promise in targeting chemoresistant CS cells, potentially improving the prognosis for patients in clinical settings.

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Microfluidic devices serve as essential tools across diverse fields like medicine, biotechnology, and chemistry, enabling advancements in analytical techniques, point-of-care diagnostics, microfluidic cell cultures, and organ-on-chip models. While polymeric microfluidics are favoured for their cost-effectiveness and ease of fabrication, their inherent hydrophobic properties necessitate surface functionalization, often post-sealing. Here, we introduce a versatile apparatus for functionalizing sealed microfluidic devices using atmospheric plasma processing, with a focus on PDMS (polydimethylsiloxane) microfluidics. Through meticulous analysis of surface properties and capillary velocity, before and after plasma treatment, along with a comparison between vacuum and atmospheric plasma functionalization methods, we demonstrate the efficacy of our approach. Subsequent experimentation within 3D PDMS microfluidic chambers, combining atmospheric pressure plasma treatment with collagen coating to facilitate mesenchymal stem cells (MSCs) growth over five days, reveals enhanced initial cell adhesion and proliferation, highlighting the potential of our method for improving cell-based applications within microfluidic systems.

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Wastewater treatment plants (WWTPs) have been implicated as direct key reservoir of both antibiotic-resistant bacteria (ARB) and antibiotic-resistant genes (ARGs) associated with human infection, as high concentrations of ARBs and ARGs have been detected in recycled hospital wastewater. Among the ARBs, the carbapenem-resistant Acinetobacter baumannii has been ranked as priority 1 (critical) pathogen by the World Health Organization (WHO), due to its overwhelming burden on public health. Therefore, this study is aimed at investigating non-thermal plasma (NTP) technology as an alternative disinfection step to inactivate this bacterium and its ARGs. Culture-based method and PCR were employed in confirming the carbapenem resistance gene blaNDM-1 in A. baumannii (BAA 1605). Suspension of carbapenem-resistant A. baumannii (24 h culture) was prepared from the confirmed isolate and subjected to plasma treatment at varying time intervals (3 min, 6 min, 9 min, 12 min, and 15 min) in triplicates. The plasma-treated samples were evaluated for re-growth and the presence of the resistance gene. The treatment resulted in a 1.13 log reduction after 3 min and the highest log reduction of ≥ 8 after 15 min, and the results also showed that NTP was able to inactivate the blaNDM-1 gene. The log reduction and gel image results suggest that plasma disinfection has a great potential to be an efficient tertiary treatment step for WWTPs.

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Introduction. Aspergillus flavus and Fusarium keratoplasticum are common causative pathogens of fungal keratitis (FK), a severe corneal disease associated with significant morbidity and vision loss. Escalating incidence of antifungal resistance to available antifungal drugs poses a major challenge to FK treatment. Cold atmospheric plasma (CAP) is a pioneering nonpharmacologic antimicrobial intervention that has demonstrated potential as a broad-spectrum antifungal treatment.Gap statement. Previous research highlights biofilm-associated resistance as a critical barrier to effective FK treatment. Although CAP has shown promise against various fungal infections, its efficacy against biofilm and conidial forms of FK pathogens remains inadequately explored.Aim. This study aims to investigate the antifungal efficacy of CAP against clinical fungal keratitis isolates of A. flavus and F. keratoplasticum in vitro.Methodology. Power parameters (22-27 kVpp, 300-400 Hz and 20-80 mA) of a dielectric barrier discharge CAP device were optimized for inactivation of A. flavus biofilms. Optimal applied voltage and total current were applied to F. keratoplasticum biofilms and conidial suspensions of A. flavus and F. keratoplasticum. The antifungal effect of CAP treatment was investigated by evaluating fungal viability through means of metabolic activity, c.f.u. enumeration (c.f.u. ml-1) and biofilm formation.Results. For both fungal species, CAP exhibited strong time-dependent inactivation, achieving greater than 80 % reduction in metabolic activity and c.f.u. ml-1 within 300 s or less, and complete inhibition after 600 s of treatment.Conclusion. Our findings indicate that CAP is a promising broad-spectrum antifungal intervention. CAP treatment effectively reduces fungal viability in both biofilm and conidial suspension cultures of A. flavus and F. keratoplasticum, suggesting its potential as an alternative treatment strategy for fungal keratitis.

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