RESUMEN
We have developed a micro electromechanical systems (MEMS) device which enables plasma treatment for cells cultured in media. The device, referred to as the plasma-on-chip, comprises microwells and microplasma sources fabricated together in a single chip. The microwells have through-holes between the microwells and microplasma sources. Each microplasma source is located on the backside of each microwells. The reactive components generated by the microplasma sources pass through the through-holes and reach cells cultured in the microwells. In this study, a plasma-on-chip device was modified for a stable plasma treatment. The use of a dielectric barrier discharge (DBD) technique allowed a stable plasma treatment up to 3 min. The plasma-on-chip with the original electrode configuration typically had the maximum stable operation time of around 1 min. Spectral analysis of the plasma identified reactive species such as O and OH radicals that can affect the activity of cells. Plasma treatment was successfully performed on yeast (Saccharomyces cerevisiae) and green algae (Chlorella) cells. While no apparent change was observed with yeast, the treatment degraded the activity of the Chlorella cells and decreased their fluorescence. The device has the potential to help understand interactions between plasma and cells.
Asunto(s)
Chlorella/citología , Frío , Sistemas Microelectromecánicos , Microelectrodos , Gases em Plasma/química , Saccharomyces cerevisiae/citología , Presión Atmosférica , Cromo/química , Medios de Cultivo , Diseño de Equipo , Oro/química , Radical Hidroxilo/química , Oxígeno/química , Silicio/químicaRESUMEN
Low-temperature plasma (LTP) treatment promotes blood clot formation by stimulation of the both platelet aggregation and coagulation factors. However, the appearance of a membrane-like structure in clots after the treatment is controversial. Based on our previous report that demonstrated characteristics of the form of coagulation of serum proteins induced by LTP treatment, we sought to determine whether treatment with two plasma instruments, namely BPC-HP1 and PN-110/120TPG, formed clots only from red blood cells (RBCs). LTP treatment with each device formed clots from whole blood, whereas LTP treatment with BPC-HP1 formed clots in phosphate-buffered saline (PBS) containing 2 × 10(9)/mL RBCs. Light microscopic analysis results showed that hemolysis formed clots consisting of materials with membrane-like structures from both whole blood and PBS-suspended RBCs. Moreover, electron microscopic analysis results showed a monotonous material with high electron density in the formed clots, presenting a membrane-like structure. Hemolysis disappeared with the decrease in the current through the targets contacting with the plasma flare and clot formation ceased. Taken together, our results and those of earlier studies present two types of blood clot formation, namely presence or absence of hemolysis capability depending on the current through the targets.
Asunto(s)
Coagulación Sanguínea , Eritrocitos/citología , Gases em Plasma , Plaquetas/metabolismo , Frío , Electrones , Membrana Eritrocítica/química , Hemoglobinas/química , Hemólisis , Humanos , Luz , Fosfatos/química , Sales (Química)/químicaRESUMEN
Over the past few years, the application of cold atmospheric plasma (CAP) in medicine has developed into an innovative field of research of rapidly growing importance. One promising new medical application of CAP is cancer treatment. Different studies revealed that CAP may potentially affect the cell cycle and cause cell apoptosis or necrosis in tumor cells dependent on the CAP device and doses. In this study, we used a novel hand-held and battery-operated CAP device utilizing the surface micro discharge (SMD) technology for plasma production in air and consequently analysed dose-dependent CAP treatment effects on melanoma cells. After 2 min of CAP treatment, we observed irreversible cell inactivation. Phospho-H2AX immunofluorescence staining and Flow cytometric analysis demonstrated that 2 min of CAP treatment induces DNA damage, promotes induction of Sub-G1 phase and strongly increases apoptosis. Further, protein array technology revealed induction of pro-apoptotic events like p53 and Rad17 phosphorylation of Cytochrome c release and activation of Caspase-3. Interestingly, using lower CAP doses with 1 min of treatment, almost no apoptosis was observed but long-term inhibition of proliferation. H3K9 immunofluorescence, SA-ß-Gal staining and p21 expression revealed that especially these low CAP doses induce senescence in melanoma cells. In summary, we observed differences in induction of apoptosis or senescence of tumor cells in respond to different CAP doses using a new CAP device. The mechanism of senescence with regard to plasma therapy was so far not described previously and is of great importance for therapeutic application of CAP.
Asunto(s)
Melanoma/terapia , Gases em Plasma/uso terapéutico , Neoplasias Cutáneas/terapia , Apoptosis , Línea Celular Tumoral , Fragmentación del ADN , Diseño de Equipo , Puntos de Control de la Fase G1 del Ciclo Celular , Humanos , Melanoma/metabolismo , Melanoma/patología , Neoplasias Cutáneas/metabolismo , Neoplasias Cutáneas/patologíaRESUMEN
Cold atmospheric plasma has already been shown to decrease the bacterial load in chronic wounds. However, until now it is not yet known if plasma treatment can also improve wound healing. We aimed to assess the impact of cold atmospheric argon plasma on the process of donor site healing. Forty patients with skin graft donor sites on the upper leg were enrolled in our study. The wound sites were divided into two equally sized areas that were randomly assigned to receive either plasma treatment or placebo (argon gas) for 2 minutes. Donor site healing was evaluated independently by two blinded dermatologists, who compared the wound areas with regard to reepithelialization, blood crusts, fibrin layers, and wound surroundings. From the second treatment day onwards, donor site wound areas treated with plasma (n = 34) showed significantly improved healing compared with placebo-treated areas (day 1, p = 0.25; day 2, p = 0.011; day 3, p < 0.001; day 4, p < 0.001; day 5, p = 0.004; day 6, p = 0.008; day 7, p = 0.031). Positive effects were observed in terms of improved reepithelialization and fewer fibrin layers and blood crusts, whereas wound surroundings were always normal, independent of the type of treatment. Wound infection did not occur in any of the patients, and no relevant side effects were observed. Both types of treatment were well tolerated. The mechanisms contributing to these clinically observed effects should be further investigated.
Asunto(s)
Coagulación con Plasma de Argón , Trasplante de Piel/métodos , Piel/patología , Cicatrización de Heridas , Infección de Heridas/patología , Heridas y Lesiones/patología , Adulto , Anciano , Anciano de 80 o más Años , Carga Bacteriana , Enfermedad Crónica , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto , Estudios Prospectivos , Repitelización/inmunología , Piel/inmunología , Piel/lesiones , Factores de Tiempo , Resultado del Tratamiento , Cicatrización de Heridas/inmunología , Infección de Heridas/inmunología , Infección de Heridas/prevención & control , Heridas y Lesiones/inmunología , Heridas y Lesiones/terapiaRESUMEN
Physical cold atmospheric surface microdischarge (SMD) plasma operating in ambient air has promising properties for the sterilization of sensitive medical devices where conventional methods are not applicable. Furthermore, SMD plasma could revolutionize the field of disinfection at health care facilities. The antimicrobial effects on Gram-negative and Gram-positive bacteria of clinical relevance, as well as the fungus Candida albicans, were tested. Thirty seconds of plasma treatment led to a 4 to 6 log(10) CFU reduction on agar plates. C. albicans was the hardest to inactivate. The sterilizing effect on standard bioindicators (bacterial endospores) was evaluated on dry test specimens that were wrapped in Tyvek coupons. The experimental D(23)(°)(C) values for Bacillus subtilis, Bacillus pumilus, Bacillus atrophaeus, and Geobacillus stearothermophilus were determined as 0.3 min, 0.5 min, 0.6 min, and 0.9 min, respectively. These decimal reduction times (D values) are distinctly lower than D values obtained with other reference methods. Importantly, the high inactivation rate was independent of the material of the test specimen. Possible inactivation mechanisms for relevant microorganisms are briefly discussed, emphasizing the important role of neutral reactive plasma species and pointing to recent diagnostic methods that will contribute to a better understanding of the strong biocidal effect of SMD air plasma.
Asunto(s)
Atmósfera/química , Bacterias/efectos de los fármacos , Candida albicans/efectos de los fármacos , Gases em Plasma/farmacología , Esporas Bacterianas/efectos de los fármacos , Esterilización/métodos , Recuento de Colonia Microbiana , Gases em Plasma/análisis , Espectrofotometría UltravioletaRESUMEN
Candida albicans is one of the main species able to form a biofilm on almost any surface, causing both skin and superficial mucosal infections. The worldwide increase in antifungal resistance has led to a decrease in the efficacy of standard therapies, prolonging treatment time and increasing health care costs. Therefore, the aim of this work was to demonstrate the applicability of atmospheric plasma at room temperature for inactivating C. albicans growing in biofilms without thermally damaging heat-sensitive materials. This so-called cold atmospheric plasma is produced by applying high voltage to accelerate electrons, which ionize the surrounding air, leading to the production of charged particles, reactive species, and photons. A newly developed plasma device was used, which exhibits a large plasma-generating surface area of 9 by 13 cm (117 cm(2)). Different time points were selected to achieve an optimum inactivation efficacy range of ≥3 log(10) to 5 log(10) reduction in CFU per milliliter, and the results were compared with those of 70% ethanol. The results obtained show that contact-free antifungal inactivation of Candida biofilms by cold atmospheric plasma is a promising tool for disinfection of surfaces (and items) in both health care settings and the food industry, where ethanol disinfection should be avoided.
Asunto(s)
Aire , Antifúngicos/farmacología , Biopelículas/efectos de los fármacos , Candida albicans/efectos de los fármacos , Gases em Plasma/farmacología , Candida albicans/fisiología , Recuento de Colonia Microbiana , Viabilidad Microbiana/efectos de los fármacos , TemperaturaRESUMEN
In this study we investigated the sensitivity of Deinococcus radiodurans to contact-free cold atmospheric plasma treatment as part of a project to establish new efficient procedures for disinfection of inanimate surfaces. The Gram-positive D. radiodurans is one of the most resistant microorganisms worldwide. Stationary phases of D. radiodurans were exposed to cold atmospheric plasma for different time intervals or to ultraviolet C (UVC) radiation at dose rates of 0.001-0.0656 J cm⻲, respectively. A methicillin-resistant Staphylococcus aureus strain (MRSA) served as control for Gram-positive bacteria. The surface microdischarge plasma technology was used for generation of cold atmospheric plasma. A plasma discharge was ignited using ambient air. Surprisingly, D. radiodurans was sensitive to the cold atmospheric plasma treatment in the same range as the MRSA strain. Survival of both bacteria decreased with increasing plasma exposure times up to 6 log10 cycles (>99.999 %) within 20 s of plasma treatment. In contrast, UVC radiation of both bacteria demonstrated that D. radiodurans was more resistant to UVC treatment than MRSA. Cold atmospheric plasma seems to be a promising tool for industrial and clinical purposes where time-saving is a critical point to achieve efficient disinfection of inanimate surfaces and where protection from corrosive materials is needed.
Asunto(s)
Deinococcus/efectos de los fármacos , Desinfección/métodos , Gases em Plasma/farmacología , Deinococcus/citología , Deinococcus/efectos de la radiación , Staphylococcus aureus Resistente a Meticilina/citología , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/efectos de la radiación , Viabilidad Microbiana/efectos de los fármacos , Viabilidad Microbiana/efectos de la radiación , Rayos UltravioletaRESUMEN
Non-thermal atmospheric-pressure plasma has been used for biological applications, including sterilization and stimulation of cell growth and differentiation. Here, we demonstrate that plasma exposure influences the differentiation pattern of human induced pluripotent stem cells (hiPSCs). We treated hiPSCs with dielectric barrier-discharge air plasma and found an exposure dose that does not kill hiPSCs. Immunohistochemical staining for E-CADHERIN showed that the exposure affected cell-cell attachment and doubled the average size of the hiPSCs. Analysis of mRNAs in embryoid bodies (EBs) from plasma-treated hiPSCs revealed repression of ectoderm genes, including WNT1, and increased expression of mesoderm genes. Importantly, hiPSCs deficient in DNA repair only displayed minimal damage after plasma exposure. Collectively, our results suggest that plasma treatment can be another tool for directing the fate of pluripotent stem cells without disrupting their genomic integrity.
RESUMEN
As a result of both the better understanding of complex plasma phenomena and the development of new plasma sources in the past few years, plasma medicine has developed into an innovative field of research showing high potential. While thermal plasmas have long been used in various medical fields (for instance for cauterization and sterilization of medical instruments), current research mainly focuses on application of non-thermal plasmas. Experiments show that cold atmospheric plasmas (CAPs) allow efficient, contact-free and painless disinfection, even in microscopic openings, without damaging healthy tissue. Plasmas influence biochemical processes and offer new possibilities for the selective application of individually designable medically active substances. In dermatology, new horizons are being opened for wound healing, tissue regeneration, therapy of skin infections, and probably many more diseases. First clinical trials show the efficacy and tolerability of plasma in treating infected chronic wounds. A major task will be the introduction of plasma into clinical medicine and, simultaneously, the further investigation of the mechanisms of action of plasma at the cellular level.
Asunto(s)
Dermatología/tendencias , Desinfección/tendencias , Gases em Plasma/uso terapéutico , Regeneración de la Piel con Plasma/tendencias , Enfermedades de la Piel/terapia , Animales , HumanosRESUMEN
OBJECTIVE: Ear, nose and throat infections are among the most common reasons for absence from work. They are usually caused by various bacteria like Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae and Streptococcus pyogenes. Cold atmospheric plasma (CAP) can effectively eliminate even multi-resistant bacteria and has no cytotoxic or mutagenic effects on the mucosa when applied for less than 60s. Aim of the study was to evaluate the effects of CAP on common ENT bacteria and on the mucosa of the upper aerodigestive tract. METHODS: The bactericidal effects of CAP against the bacteria most commonly causing ENT infections were investigated using the colony-forming units assay (CFU) on a Müller-Hinton agar plate after applying CAP for 30, 60, 90 and 120s. To evaluate the interaction of CAP with mucosal cells, 3D mini organ cultures were treated for up to 180s, after which cell viability and necrosis induction were evaluated. RESULTS: Treatment with CAP for 60s or longer induced at least a 3-log10 reduction in the bacterial load (> 99.9%). Treatment times shorter than 60s had only slight cytotoxic effects on cell viability and necrosis whereas treatment times above 60s showed a fast increase of cytotoxic side effects. CONCLUSION: CAP exhibited strong bactericidal effects on the most common ENT pathogens. Treatment times of up to 60s showed only minimal adverse reactions in healthy mucosa. CAP could be a promising new therapeutic modality for ENT infections.
Asunto(s)
Haemophilus influenzae/efectos de los fármacos , Otitis/microbiología , Gases em Plasma/farmacología , Mucosa Respiratoria/efectos de los fármacos , Infecciones del Sistema Respiratorio/microbiología , Staphylococcus aureus/efectos de los fármacos , Streptococcus pneumoniae/efectos de los fármacos , Streptococcus pyogenes/efectos de los fármacos , Adulto , Supervivencia Celular/efectos de los fármacos , Recuento de Colonia Microbiana , Femenino , Humanos , Masculino , Persona de Mediana Edad , Necrosis , Técnicas de Cultivo de Órganos , Orofaringe , Faringitis/microbiología , Mucosa Respiratoria/patología , Rinitis/microbiología , Adulto JovenRESUMEN
Cold atmospheric plasma (CAP) has been gaining increasing interest as a new approach for the treatment of skin diseases or wounds. Although this approach has demonstrated promising antibacterial activity, its exact mechanism of action remains unclear. This study explored in vitro and in vivo whether CAP influences gene expression and molecular mechanisms in keratinocytes. Our results revealed that a 2 min CAP treatment using the MicroPlaSter ß in analogy to the performed clinical studies for wound treatment induces expression of IL-8, TGF-ß1, and TGF-ß2. In vitro and in vivo assays indicated that keratinocyte proliferation, migration, and apoptotic mechanisms were not affected by the CAP treatment under the applied conditions. Further, we observed that antimicrobial peptides of the ß-defensin family are upregulated after CAP treatment. In summary, our results suggest that a 2 min application of CAP induces gene expression of key regulators important for inflammation and wound healing without causing proliferation, migration or cell death in keratinocytes. The induction of ß-defensins in keratinocytes describes an absolutely new plasma strategy. Activation of antimicrobial peptides supports the well-known antibacterial effect of CAP treatment, whereas the mechanism of ß-defensin activation by CAP is not investigated so far.
Asunto(s)
Apoptosis/efectos de los fármacos , Atmósfera/química , Citocinas/metabolismo , Queratinocitos/citología , Queratinocitos/metabolismo , Gases em Plasma/farmacología , beta-Defensinas/metabolismo , Citocinas/genética , Fibrosis , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Inflamación/metabolismo , Queratinocitos/efectos de los fármacos , Piel/citología , Piel/patología , Cicatrización de Heridas/efectos de los fármacos , beta-Defensinas/genéticaRESUMEN
Head and neck squamous cell cancer (HNSCC) is the 7th most common cancer worldwide. Despite the development of new therapeutic agents such as monoclonal antibodies, prognosis did not change for the last decades. Cold atmospheric plasma (CAP) presents the most promising new technology in cancer treatment. In this study the efficacy of a surface micro discharging (SMD) plasma device against two head and neck cancer cell lines was proved. Effects on the cell viability, DNA fragmentation and apoptosis induction were evaluated with the MTT assay, alkaline microgel electrophoresis (comet assay) and Annexin-V/PI staining. MTT assay revealed that the CAP treatment markedly decreases the cell viability for all tested treatment times (30, 60, 90, 120 and 180 s). IC 50 was reached within maximal 120 seconds of CAP treatment. Comet assay analysis showed a dose dependent high DNA fragmentation being one of the key players in anti-cancer activity of CAP. Annexin-V/PI staining revealed induction of apoptosis in CAP treated HNSCC cell lines but no significant dose dependency was seen. Thus, we confirmed that SMD Plasma technology is definitely a promising new approach on cancer treatment.
Asunto(s)
Carcinoma de Células Escamosas/terapia , Neoplasias de Cabeza y Cuello/terapia , Gases em Plasma/uso terapéutico , Apoptosis/efectos de la radiación , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Supervivencia Celular/efectos de la radiación , Fragmentación del ADN/efectos de la radiación , Neoplasias de Cabeza y Cuello/patología , HumanosRESUMEN
Cold atmospheric plasma science is an innovative upcoming technology for the medical sector. The plasma composition and subsequent effects on cells, tissues and pathogens can vary enormously depending on the plasma source, the plasma settings and the ambient conditions. Cold atmospheric plasmas consist of a highly reactive mix of ions and electrons, reactive molecules, excited species, electric fields and to some extent also UV radiation. In the last year, this partly ionized gas has been demonstrated to have a broad antimicrobial activity, while resistance and resistance development are unlikely. Furthermore, recent research has indicated that plasmas also have a strong influence on various cell lines and cell functions, including anticancer properties. This review summarizes the major plasma designs available and their main benefits, as well as assessing possible risks of this new technology.
Asunto(s)
Equipos y Suministros , Gases em Plasma/uso terapéutico , Atmósfera , Equipos y Suministros/efectos adversos , Equipos y Suministros/clasificación , Humanos , Gases em Plasma/efectos adversos , Medición de RiesgoRESUMEN
AIM: Cold atmospheric plasma (CAP) has already proven efficient at disinfection of microorganisms including biofilms. The objective of the present study is to assess the efficacy of CAP against the dermatophytes Trichophyton rubrum and Microsporum canis in vitro. MATERIALS & METHODS: T. rubrum and M. canis were exposed to CAP for different treatment times and time intervals in vitro. Treatment with ciclopirox olamine or UVC radiation (0.120 J/cm(2)) served as controls. CAP was generated by the surface microdischarge technology. Fungal colony growth was measured upon CAP treatment. RESULTS: Repeated daily CAP treatments of 10 min demonstrated an inhibition of growth during the treatment period of 9 days. Single CAP treatment sessions for 5, 8 and 10 min, as well as treatments for 5 or 8 min daily, resulted in less fungal growth inhibition. UVC radiation treatment failed, but not ciclopirox olamine. CONCLUSION: CAP shows promising potential for future application in the treatment of dermatophyte infections.
Asunto(s)
Desinfectantes/farmacología , Viabilidad Microbiana/efectos de los fármacos , Microsporum/efectos de los fármacos , Gases em Plasma/farmacología , Trichophyton/efectos de los fármacos , Recuento de Colonia Microbiana , Relación Dosis-Respuesta a Droga , Factores de TiempoRESUMEN
Cold atmospheric plasma (CAP) has the potential to interact with tissue or cells leading to fast, painless and efficient disinfection and furthermore has positive effects on wound healing and tissue regeneration. For clinical implementation it is necessary to examine how CAP improves wound healing and which molecular changes occur after the CAP treatment. In the present study we used the second generation MicroPlaSter ß® in analogy to the current clinical standard (2 min treatment time) in order to determine molecular changes induced by CAP using in vitro cell culture studies with human fibroblasts and an in vivo mouse skin wound healing model. Our in vitro analysis revealed that the CAP treatment induces the expression of important key genes crucial for the wound healing response like IL-6, IL-8, MCP-1, TGF-ß1, TGF-ß2, and promotes the production of collagen type I and alpha-SMA. Scratch wound healing assays showed improved cell migration, whereas cell proliferation analyzed by XTT method, and the apoptotic machinery analyzed by protein array technology, was not altered by CAP in dermal fibroblasts. An in vivo wound healing model confirmed that the CAP treatment affects above mentioned genes involved in wound healing, tissue injury and repair. Additionally, we observed that the CAP treatment improves wound healing in mice, no relevant side effects were detected. We suggest that improved wound healing might be due to the activation of a specified panel of cytokines and growth factors by CAP. In summary, our in vitro human and in vivo animal data suggest that the 2 min treatment with the MicroPlaSter ß® is an effective technique for activating wound healing relevant molecules in dermal fibroblasts leading to improved wound healing, whereas the mechanisms which contribute to these observed effects have to be further investigated.
Asunto(s)
Presión Atmosférica , Frío , Gases em Plasma/farmacología , Cicatrización de Heridas/efectos de los fármacos , Actinas/genética , Animales , Apoptosis/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Colágeno/biosíntesis , Citocinas/metabolismo , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Péptidos y Proteínas de Señalización Intercelular/metabolismo , RatonesRESUMEN
Glioblastoma (GBM) is the most common and aggressive brain tumor in adults. Despite multimodal treatments including surgery, chemotherapy and radiotherapy the prognosis remains poor and relapse occurs regularly. The alkylating agent temozolomide (TMZ) has been shown to improve the overall survival in patients with malignant gliomas, especially in tumors with methylated promoter of the O6-methylguanine-DNA-methyltransferase (MGMT) gene. However, intrinsic and acquired resistance towards TMZ makes it crucial to find new therapeutic strategies aimed at improving the prognosis of patients suffering from malignant gliomas. Cold atmospheric plasma is a new auspicious candidate in cancer treatment. In the present study we demonstrate the anti-cancer properties of different dosages of cold atmospheric plasma (CAP) both in TMZ-sensitive and TMZ-resistant cells by proliferation assay, immunoblotting, cell cycle analysis, and clonogenicity assay. Importantly, CAP treatment restored the responsiveness of resistant glioma cells towards TMZ therapy. Concomitant treatment with CAP and TMZ led to inhibition of cell growth and cell cycle arrest, thus CAP might be a promising candidate for combination therapy especially for patients suffering from GBMs showing an unfavorable MGMT status and TMZ resistance.
Asunto(s)
Atmósfera , Neoplasias Encefálicas/tratamiento farmacológico , Resistencia a Antineoplásicos/efectos de los fármacos , Glioma/tratamiento farmacológico , Gases em Plasma/uso terapéutico , Adulto , Neoplasias Encefálicas/patología , Muerte Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Daño del ADN , Dacarbazina/análogos & derivados , Dacarbazina/farmacología , Dacarbazina/uso terapéutico , Relación Dosis-Respuesta a Droga , Fase G2/efectos de los fármacos , Glioma/patología , Humanos , Gases em Plasma/farmacología , Temozolomida , Ensayo de Tumor de Célula MadreRESUMEN
In the last twenty years new antibacterial agents approved by the U.S. FDA decreased whereas in parallel the resistance situation of multi-resistant bacteria increased. Thus, community and nosocomial acquired infections of resistant bacteria led to a decrease in the efficacy of standard therapy, prolonging treatment time and increasing healthcare costs. Therefore, the aim of this work was to demonstrate the applicability of cold atmospheric plasma for decolonisation of Gram-positive (Methicillin-resistant Staphylococcus aureus (MRSA), Methicillin-sensitive Staphylococcus aureus) and Gram-negative bacteria (E. coli) using an ex vivo pig skin model. Freshly excised skin samples were taken from six month old female pigs (breed: Pietrain). After application of pure bacteria on the surface of the explants these were treated with cold atmospheric plasma for up to 15 min. Two different plasma devices were evaluated. A decolonisation efficacy of 3 log(10) steps was achieved already after 6 min of plasma treatment. Longer plasma treatment times achieved a killing rate of 5 log(10) steps independently from the applied bacteria strains. Histological evaluations of untreated and treated skin areas upon cold atmospheric plasma treatment within 24 h showed no morphological changes as well as no significant degree of necrosis or apoptosis determined by the TUNEL-assay indicating that the porcine skin is still vital. This study demonstrates for the first time that cold atmospheric plasma is able to very efficiently kill bacteria applied to an intact skin surface using an ex vivo porcine skin model. The results emphasize the potential of cold atmospheric plasma as a new possible treatment option for decolonisation of human skin from bacteria in patients in the future without harming the surrounding tissue.
Asunto(s)
Desinfección/métodos , Escherichia coli/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Gases em Plasma/farmacología , Piel/microbiología , Staphylococcus aureus/efectos de los fármacos , Animales , Femenino , Técnicas Histológicas , Etiquetado Corte-Fin in Situ , Técnicas In Vitro , Sus scrofa , Factores de TiempoRESUMEN
Non-thermal (low-temperature) physical plasma is under intensive study as an alternative approach to control superficial wound and skin infections when the effectiveness of chemical agents is weak due to natural pathogen or biofilm resistance. The purpose of this study was to test the individual susceptibility of pathogenic bacteria to non-thermal argon plasma and to measure the effectiveness of plasma treatments against bacteria in biofilms and on wound surfaces. Overall, Gram-negative bacteria were more susceptible to plasma treatment than Gram-positive bacteria. For the Gram-negative bacteria Pseudomonas aeruginosa, Burkholderia cenocepacia and Escherichia coli, there were no survivors among the initial 10(5) c.f.u. after a 5 min plasma treatment. The susceptibility of Gram-positive bacteria was species- and strain-specific. Streptococcus pyogenes was the most resistant with 17â% survival of the initial 10(5) c.f.u. after a 5 min plasma treatment. Staphylococcus aureus had a strain-dependent resistance with 0 and 10â% survival from 10(5) c.f.u. of the Sa 78 and ATCC 6538 strains, respectively. Staphylococcus epidermidis and Enterococcus faecium had medium resistance. Non-ionized argon gas was not bactericidal. Biofilms partly protected bacteria, with the efficiency of protection dependent on biofilm thickness. Bacteria in deeper biofilm layers survived better after the plasma treatment. A rat model of a superficial slash wound infected with P. aeruginosa and the plasma-sensitive Staphylococcus aureus strain Sa 78 was used to assess the efficiency of argon plasma treatment. A 10 min treatment significantly reduced bacterial loads on the wound surface. A 5-day course of daily plasma treatments eliminated P. aeruginosa from the plasma-treated animals 2 days earlier than from the control ones. A statistically significant increase in the rate of wound closure was observed in plasma-treated animals after the third day of the course. Wound healing in plasma-treated animals slowed down after the course had been completed. Overall, the results show considerable potential for non-thermal argon plasma in eliminating pathogenic bacteria from biofilms and wound surfaces.