Improving the Stability of Amino-Containing Plasma Polymer Films in Aqueous Environments.

Plasma polymer films that contain amine groups (NH2-PPFs) are known to degrade over time, particularly in aqueous environments. To reduce such aging effects, a vertical chemical gradient regarding the amine group density was explored ranging over a few nanometers at the coating surface. The gradient-containing nanofilms were formed in low-pressure plasma by tuning plasma conditions while keeping the plasma "switched on". The coating process started with a more cross-linked NH2-PPF (70 W, 4:7 NH3/C2H4), followed by the deposition of a few nanometers of a less cross-linked yet more functional NH2-PPF (50 W, 7:7 NH3/C2H4). Characterization of the prepared gradient coatings showed that the chemical composition depends on the NH3/C2H4 gas flow ratio, as observed by different analytical methods: plasma diagnostics during deposition and depth profiling analyses of the deposited coating. Finally, surface chemistry was analyzed during air and water aging, showing a similar aging process of the NH2-PPF single layer and NH2-PPF with a vertical chemical gradient in air, while the stability of the gradient coating was found to be enhanced under aqueous conditions maintaining an [NH2]/[C] amount of ∼1%.

Risk assessment of a cold argon plasma jet in respect to its mutagenicity.

Cold atmospheric pressure plasmas represent a favorable option for the treatment of heat sensitive materials and human or animal tissue. Beneficial effects have been documented in a variety of medical conditions, e.g., in the treatment of chronic wounds. It is assumed that the main mechanism of the plasma's efficacy is mediated by a stimulating dissipation of energy via radiation and/or chemical energy. Although no evidence on undesired side effects of a plasma treatment has yet been presented, skepticism toward the safety of the exposure to plasma is present. However, only little data regarding the mutagenic potential of this new treatment option is available. Accordingly, we investigated the mutagenic potential of an argon plasma jet (kinpen) using different testing systems in accordance with ISO norms and multiple cell lines: a HPRT1 mutation assay, a micronucleus formation assay, and a colony formation assay. Moderate plasma treatment up to 180 s did not increase genotoxicity in any assay or cell type investigated. We conclude that treatment with the argon plasma jet kinpen did not display a mutagenic potential under the test conditions applied and may from this perspective be regarded as safe for the use in biomedical applications.

Hydrogen peroxide: A central player in physical plasma-induced oxidative stress in human blood cells.

Plasma medicine is an interdisciplinary field and recent clinical studies showed benefits of topical plasma application to chronic wounds. Whereas most investigations have focused on plasma-skin cell interaction, immune cells are omnipresent in most tissues as well. They not only elicit specific immune responses but also regulate inflammation, which is central in healing and regeneration. Plasma generates short-lived radicals and species in the gas phase. Mechanisms of plasma-cell interactions are not fully understood but it is hypothesized that reactive oxygen and nitrogen species (RONS) mediate effects of plasma on cells. In this study human blood cells were investigated after cold atmospheric plasma treatment with regard to oxidation and viability. Plasma generates hydrogen peroxide (H2O2) and the responses were similar in cells treated with concentration-matched H2O2. Both treatments gave an equivalent reduction in viability and this was completely abrogated if catalase was added prior to plasma exposure. Further, five oxidation probes were utilized and fluorescence increase was observed in plasma-treated cells. Dye-dependent addition of catalase diminished most but not all of the probe fluorescence, assigning H2O2 a dominant but not exclusive role in cellular oxidation by plasma. Investigations for other species revealed generation of nitrite and formation of 3-nitrotyrosine but not 3-chlorotyrosine after plasma treatment indicating presence of RNS which may contribute to cellular redox changes observed. Together, these results will help to clarify how oxidative stress associates with physical plasma treatment in wound relevant cells.

Efficacy of different carrier gases for barrier discharge plasma generation compared to chlorhexidine on the survival of Pseudomonas aeruginosa embedded in biofilm in vitro.

Because of its antimicrobial properties, nonthermal plasma could serve as an alternative to chemical antisepsis in wound treatment. Therefore, this study investigated the inactivation of biofilm-embedded Pseudomonas aeruginosa SG81 by a surface barrier-discharged (SBD) plasma for 30, 60, 150 and 300 s. In order to optimize the efficacy of the plasma, different carrier gases (argon, argon admixed with 1% oxygen, and argon with increased humidity up to approx. 80%) were tested and compared against 0.1% chlorhexidine digluconate (CHG) exposure for 600 s. The antimicrobial efficacy was determined by calculating the difference between the numbers of colony-forming units (CFU) of treated and untreated biofilms. Living bacteria were distinguished from dead by fluorescent staining and confocal laser scanning microscopy. Both SBD plasmas and CHG showed significant antimicrobial effects compared to the untreated control. However, plasma treatment led to a higher antimicrobial reduction (argon plasma 4.9 log10 CFU/cm(2), argon with admixed oxygen 3 log10 CFU/cm(2), and with increased gas humidity 2.7 log10 CFU/cm(2) after 300 s) compared to CHG. In conclusion, SBD plasma is suitable as an alternative to CHG for inactivation of Pseudomonas aeruginosa embedded in biofilm. Further development of SBD plasma sources and research on the role of carrier gases and humidity may allow their clinical application for wound management in the future.

Plasma pharmacy - physical plasma in pharmaceutical applications.

During the last years the use of physical plasma for medical applications has grown rapidly. A multitude of findings about plasma-cell and plasma-tissue interactions and its possible use in therapy have been provided. One of the key findings of plasma medical basic research is that several biological effects do not result from direct plasma-cell or plasma-tissue interaction but are mediated by liquids. Above all, it was demonstrated that simple liquids like water or physiological saline, are antimicrobially active after treatment by atmospheric pressure plasma and that these effects are attributable to the generation of different low-molecular reactive species. Besides, it could be shown that plasma treatment leads to the stimulation of specific aspects of cell metabolism and to a transient and reversible increase of diffusion properties of biological barriers. All these results gave rise to think about another new and innovative field of medical plasma application. In contrast to plasma medicine, which means the direct use of plasmas on or in the living organism for direct therapeutic purposes, this field - as a specific field of medical plasma application - is called plasma pharmacy. Based on the present state of knowledge, most promising application fields of plasma pharmacy might be: plasma-based generation of biologically active liquids; plasma-based preparation, optimization, or stabilization of - mainly liquid - pharmaceutical preparations; support of drug transport across biological barriers; plasma-based stimulation of biotechnological processes.

Laser schlieren deflectometry for temperature analysis of filamentary non-thermal atmospheric pressure plasma.

The heat convection generated by micro filaments of a self-organized non-thermal atmospheric pressure plasma jet in Ar is characterized by employing laser schlieren deflectometry (LSD). It is demonstrated as a proof of principle, that the spatial and temporal changes of the refractive index n in the optical beam path related to the neutral gas temperature of the plasma jet can be monitored and evaluated simultaneously. The refraction of a laser beam in a high gradient field of n(r) with cylindrical symmetry is given for a general real refraction index profile. However, the usually applied Abel approach represents an ill-posed problem and in particular for this plasma configuration. A simple analytical model is proposed in order to minimize the statistical error. Based on that, the temperature profile, specifically the absolute temperature in the filament core, the FWHM, and the frequencies of the collective filament dynamics are obtained for non-stationary conditions. For a gas temperature of 700 K inside the filament, the presented model predicts maximum deflection angles of the laser beam of 0.3 mrad which is in accordance to the experimental results obtained with LSD. Furthermore, the experimentally obtained FWHM of the temperature profile produced by the filament at the end of capillary is (1.5 ± 0.2) mm, which is about 10 times wider than the visual radius of the filament. The obtained maximum temperature in the effluent is (450 ± 30) K and is in consistence with results of other techniques. The study demonstrates that LSD represents a useful low-cost method for monitoring the spatiotemporal behaviour of microdischarges and allows to uncover their dynamic characteristics, e.g., the temperature profile even for challenging diagnostic conditions such as moving thin discharge filaments. The method is not restricted to the miniaturized and self-organized plasma studied here. Instead, it can be readily applied to other configurations that produce measurable gradients of refractive index by local gas heating and opens new diagnostics prospects particularly for microplasmas.

Skin decontamination by low-temperature atmospheric pressure plasma jet and dielectric barrier discharge plasma.

BACKGROUND: Over the past few years, plasma medicine has become an important field in medical science. Cold plasma has proven anti-inflammatory, antimicrobial and antineoplastic effects. AIM: To test the decontamination power of two cold plasma sources [low-temperature atmospheric pressure plasma jet (APPJ) and dielectric barrier discharge plasma (DBD)] in vivo on human fingertips. METHODS: After 3, 15, 30, 60, 90, 120, 150, 180, 210 and 240 s of spot treatment with the APPJ and DBD, the log reduction factors (RFs) of physiological (PF) and artificially (AF) contaminated flora (Staphylococcus epidermidis and Micrococcus luteus) were calculated. The bacterial load was determined after counting. Tolerance (paresthesia, pain and heat) was measured using a numerical rating scale. FINDINGS: Both plasma devices led to a significant reduction in PF and AF. The maximum log reduction factors for PF were 1.3 for the DBD at 210 s and 0.8 for the APPJ at 60 s. For AF, the maximum log reduction factors were 1.7 for the DBD at 90 s and 1.4 for the APPJ at 120 s. Treatment with both devices was well tolerated. CONCLUSION: Both the APPJ and DBD were highly effective in eradicating PF and AF from the fingertips of healthy volunteers. No plasma-resistant isolates were observed. Cold plasma appears to have potential for skin disinfection. For hand hygiene purposes, plasma exposure times would need to be reduced significantly by technical means.

Comparison of the antiseptic efficacy of tissue-tolerable plasma and an octenidine hydrochloride-based wound antiseptic on human skin.

Colonization and infection of wounds represent a major reason for the impairment of tissue repair. Recently, it has been reported that tissue-tolerable plasma (TTP) is highly efficient in the reduction of the bacterial load of the skin. In the present study, the antiseptic efficacy of TTP was compared to that of octenidine hydrochloride with 2-phenoxyethanol. Both antiseptic methods proved to be highly efficient. Cutaneous treatment of the skin with octenidine hydrochloride and 2-phenoxyethanol leads to a 99% elimination of the bacteria, and 74% elimination is achieved by TTP treatment. Technical challenges with an early prototype TTP device could be held responsible for the slightly reduced antiseptic properties of TTP, compared to a standard antiseptic solution, since the manual treatment of the skin surface with a small beam of the TTP device might have led to an incomplete coverage of the treated area.

Skin disinfection by plasma-tissue interaction: comparison of the effectivity of tissue-tolerable plasma and a standard antiseptic.

Wound healing disorders frequently occur due to biofilm formation on wound surfaces requiring conscientious wound hygiene. Often, the application of conventional liquid antiseptics is not sufficient and sustainable as (1) the borders and the surrounding of chronic wounds frequently consist of sclerotic skin, impeding an effectual penetration of these products, and (2) the hair follicles representing the reservoir for bacterial recolonization of skin surfaces are not affected. Recently, it has been reported that tissue-tolerable plasma (TTP), which is used at a temperature range between 35 and 45°C, likewise has disinfecting properties. In the present study, the effectivity of TTP and a standard liquid antiseptic was compared in vitro on porcine skin. The results revealed that TTP was able to reduce the bacterial load by 94%, although the application of the liquid antiseptic remained superior as it reduced the bacteria by almost 99%. For in vivo application, however, TTP offers several advantages. On the one hand, TTP enables the treatment of sclerotic skin as well, and on the other hand, a sustainable disinfection can be realized as, obviously, also the follicular reservoir is affected by TTP.

Influence of age and sex in exhaled breath samples investigated by means of infrared laser absorption spectroscopy.

Breath gas analysis provides insight into human metabolism of healthy and ill individuals. As an innovative and non-invasive method, it opens up options to improve diagnostics, monitoring and treatment decisions. Mid-infrared laser absorption spectroscopy is utilized to detect CH(4), H(2)O, CO(2), NH(3) and CH(3)OH in exhaled human breath. An off-line approach using breath sampling by means of Tedlar bags is applied. The breath gas samples are measured within the population-based epidemiological Study of Health in Pomerania (SHIP-TREND) performed at the University of Greifswald. The study covers about 5000 adult subjects aged 20-79 years within 3 years. Besides breath gas analysis many other examinations are conducted. It is expected to find associations between distinct concentration levels of species in the exhaled breath and diseases assessed in this study. The study will establish reference values for exhaled breath components and serve as background population for case-control studies. In the long run, morbidity and mortality follow-ups will be conducted, which will answer the question whether end-expiratory breath gas components predict future diseases and death. As first results, we present data from 45 dialysis patients (23 males, 22 females) which were recruited in a preliminary study in preparation for SHIP-TREND.

Antiseptic efficacy and tolerance of tissue-tolerable plasma compared with two wound antiseptics on artificially bacterially contaminated eyes from commercially slaughtered pigs.

AIM: To compare the tissue tolerance and efficacy of two wound antiseptics with tissue-tolerable plasma (TTP) on enucleated contaminated eyes from slaughtered pigs in order to draw consequences for the use of TTP on wounds. METHOD: The corneas of extracted eyes were contaminated with Staphylococcus aureus or Pseudomonas aeruginosa. One and 10 min after application of 10% povidone (PVP)-iodine and 0.04% polyhexanide, respectively, the eyes were rinsed with inactivating solution. To test TTP, the plasma pen meandered over the eyes at a speed of 30 mm/s and a distance of 5 mm; the eyes were then rinsed with balanced salt solution. The reduction factor was calculated by the difference between the logarithm of colony-forming units in the rinse before and after antisepsis or TTP application. RESULTS: The efficacy of TTP (reduction factor 2.4-2.9) was significantly higher (p < 0.001) than that of PVP-iodine and polyhexanide (reduction factor 1.7-2.1). CONCLUSION: TTP is more effective than the tested wound antiseptics. The lack of histological damage to the eyes of slaughtered pigs would seem to make its use as a wound antiseptic a viable alternative. In contrast to antiseptics, it supplies additional energy in the form of heat, electric fields and radicals by TTP.