Search Results
Reactive species driven oxidative modifications of peptides—Tracing physical plasma liquid chemistry
2021, Wenske, Sebastian, Lackmann, Jan-Wilm, Busch, Larissa Milena, Bekeschus, Sander, von Woedtke, Thomas, Wende, Kristian
The effluence of physical plasma consists of a significant share of reactive species, which may interact with biomolecules and yield chemical modifications comparable to those of physiological processes, e.g., post-translational protein modifications (oxPTMs). Consequentially, the aim of this work is to understand the role of physical plasma-derived reactive species in the introduction of oxPTM-like modifications in proteins. An artificial peptide library consisting of ten peptides was screened against the impact of two plasma sources, the argon-driven MHz-jet kINPen and the helium-driven RF-jet COST-Jet. Changes in the peptide molecular structure were analyzed by liquid chromatography–mass spectrometry. The amino acids cysteine, methionine, tyrosine, and tryptophan were identified as major targets. The introduction of one, two, or three oxygen atoms was the most common modification observed. Distinct modification patterns were observed for nitration (+N + 2O–H), which occurred in kINPen only (peroxynitrite), and chlorination (+Cl–H) that was exclusive for the COST-Jet in the presence of chloride ions (atomic oxygen/hypochlorite). Predominantly for the kINPen, singlet oxygen-related modifications, e.g., cleavage of tryptophan, were observed. Oxidation, carbonylation, and double oxidations were attributed to the impact of hydroxyl radicals and atomic oxygen. Leading to a significant change in the peptide side chain, most of these oxPTM-like modifications affect the secondary structure of amino acid chains, and amino acid polarity/functionality, ultimately modifying the performance and stability of cellular proteins.
Influence of aerosol injection on the liquid chemistry induced by an RF argon plasma jet
2021, Sremački, Ivana, Bruno, Giuliana, Jablonowski, Helena, Leys, Christophe, Nikiforov, Anton, Wende, Kristian
A radio-frequency driven plasma jet in annular geometry coupled with an aerosol injection into the effluent is proposed for the controllable reactive oxygen species (ROS)/reactive nitrogen species (RNS) production and delivery on biological targets in the context of plasma medicine, e.g. wound care. The role of the aqueous aerosol in modulating the reactive species production is investigated by combining physical and chemical analytics. Optical emission spectroscopy, electron paramagnetic resonance spectroscopy, and a biochemical model based on cysteine as a tracer molecule have been applied, revealing that aerosol injection shifts the production of ROS from atomic and singlet oxygen toward hydroxyl radicals, which are generated in the droplets. Species generation occurred mainly at the droplets boundary layer during their transport through the effluent, leading to a limited cysteine turnover upon introduction into the aerosol solution. The subsequent delivery of unmodified cysteine molecules at a target suggested the application of the plasma source for the topical delivery of drugs, expanding the potential applicability and effectiveness. The presence of RNS was negligible regardless of aerosol injection and only traces of the downstream products nitrate and nitrate were detected. In summary, the aerosol injection into the effluent opens new avenues to control UV radiation and reactive species output for the biomedical applications of non-thermal plasma sources, reaching out toward the regulation, safety, and efficacy of targeted applications.
Development of an electrochemical sensor for in-situ monitoring of reactive species produced by cold physical plasma
2021, Nasri, Zahra, Bruno, Giuliana, Bekeschus, Sander, Weltmann, Klaus-Dieter, von Woedtke, Thomas, Wende, Kristian
The extent of clinical applications of oxidative stress-based therapies such as photodynamic therapy (PDT) or respiratory chain disruptors are increasing rapidly, with cold physical plasma (CPP) emerging as a further option. According to the current knowledge, the biological effects of CPP base on reactive oxygen and nitrogen species (RONS) relevant in cell signaling. To monitor the safety and the biological impact of the CPP, determining the local generation of RONS in the same environment in which they are going to be applied is desirable. Here, for the first time, the development of an electrochemical sensor for the simple, quick, and parallel determination of plasma-generated reactive species is described. The proposed sensor consists of a toluidine blue redox system that is covalently attached to a gold electrode surface. By recording chronoamperometry at different potentials, it is possible to follow the in-situ production of the main long-lived reactive oxygen and nitrogen species like hydrogen peroxide, nitrite, hypochlorite, and chloramine with time. The applicability of this electrochemical sensor for the in-situ assessment of reactive species in redox-based therapies is demonstrated by the precise analysis of hydrogen peroxide dynamics in the presence of blood cancer cells.