Photons and particles emitted from cold atmospheric-pressure plasma inactivate bacteria and biomolecules independently and synergistically

Cold atmospheric-pressure plasmas are currently in use in medicine as surgical tools and are being evaluated for new applications, including wound treatment and cosmetic care. The disinfecting properties of plasmas are of particular interest, given the threat of antibiotic resistance to modern medicine. Plasma effluents comprise (V)UV photons and various reactive particles, such as accelerated ions and radicals, that modify biomolecules; however, a full understanding of the molecular mechanisms that underlie plasma-based disinfection has been lacking. Here, we investigate the antibacterial mechanisms of plasma, including the separate, additive and synergistic effects of plasma-generated (V)UV photons and particles at the cellular and molecular levels. Using scanning electron microscopy, we show that plasma-emitted particles cause physical damage to the cell envelope, whereas UV radiation does not. The lethal effects of the plasma effluent exceed the zone of physical damage. We demonstrate that both plasma-generated particles and (V)UV photons modify DNA nucleobases. The particles also induce breaks in the DNA backbone. The plasma effluent, and particularly the plasma-generated particles, also rapidly inactivate proteins in the cellular milieu. Thus, in addition to physical damage to the cellular envelope, modifications to DNA and proteins contribute to the bactericidal properties of cold atmospheric-pressure plasma.

[1]  P. Schaeffer,et al.  (Symposium on Bacterial Spores: Paper II). Genetics of Sporulation in Bacillus subtilis Marburg , 1970 .

[2]  M. Hecker,et al.  SigB-dependent general stress response in Bacillus subtilis and related gram-positive bacteria. , 2007, Annual review of microbiology.

[3]  G. Thomas,et al.  Polarized Raman spectra of oriented fibers of A DNA and B DNA: anisotropic and isotropic local Raman tensors of base and backbone vibrations. , 1995, Biophysical journal.

[4]  In-Seop Lee,et al.  Sterilization using a microwave-induced argon plasma system at atmospheric pressure , 2003 .

[5]  J. Benedikt,et al.  Characterization of the effluent of a He/O2 microscale atmospheric pressure plasma jet by quantitative molecular beam mass spectrometry , 2010 .

[6]  M. Hecker,et al.  The Clp Proteases of Bacillus subtilisAre Directly Involved in Degradation of Misfolded Proteins , 2000, Journal of bacteriology.

[7]  Sun Choi,et al.  Oxidative modifications of glyceraldehyde-3-phosphate dehydrogenase play a key role in its multiple cellular functions. , 2009, The Biochemical journal.

[8]  J. Michel,et al.  Symposium on bacterial spores: II. Genetics of sporulation in Bacillus subtilis Marburg. , 1970, Journal of Applied Bacteriology.

[9]  N. Chebotareva,et al.  Kinetics of aggregation of UV-irradiated glyceraldehyde-3-phosphate dehydrogenase from rabbit skeletal muscle. Effect of agents possessing chaperone-like activity. , 2012, Biophysical chemistry.

[10]  R. Foest,et al.  Vacuum Ultraviolet (VUV) Emission of an Atmospheric Pressure Plasma Jet (μ-APPJ) Operated in Helium-Oxygen Mixtures in Ambient Air , 2009 .

[11]  J. Helmann,et al.  Promoter Recognition by Bacillus subtilisςW: Autoregulation and Partial Overlap with the ςX Regulon , 1998 .

[12]  M Landthaler,et al.  Successful and safe use of 2 min cold atmospheric argon plasma in chronic wounds: results of a randomized controlled trial , 2012, The British journal of dermatology.

[13]  E. Oliveros,et al.  Hydrogen peroxide evolution during V-UV photolysis of water. , 2005, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[14]  Akira Mizuno,et al.  Biological Evaluation of DNA Damage in Bacteriophages Inactivated by Atmospheric Pressure Cold Plasma , 2010 .

[15]  D. Bol,et al.  Characterization of an inducible oxidative stress system in Bacillus subtilis , 1990, Journal of bacteriology.

[16]  W. Peticolas,et al.  Conformational dependence of the Raman scattering intensities from polynucleotides. III. Order‐disorder changes in helical structures , 1971, Biopolymers.

[17]  X. Xing,et al.  Novel mutation breeding method for Streptomyces avermitilis using an atmospheric pressure glow discharge plasma , 2010, Journal of applied microbiology.

[18]  Thomas von Woedtke,et al.  Estimation of Possible Mechanisms of Escherichia coli Inactivation by Plasma Treated Sodium Chloride Solution , 2011 .

[19]  P. Schroeder,et al.  Irradiation of GAPDH: a model for environmentally induced protein damage , 2007, Biological chemistry.

[20]  C. Kieda,et al.  ROS implication in a new antitumor strategy based on non‐thermal plasma , 2012, International journal of cancer.

[21]  M. Rong,et al.  Main Species and Physicochemical Processes in Cold Atmospheric‐pressure He + O2 Plasmas , 2010 .

[22]  S. Reuter,et al.  Atomic oxygen formation in a radio-frequency driven micro-atmospheric pressure plasma jet , 2010 .

[23]  H. Tanii,et al.  Effect of acrylamide and related compounds on glycolytic enzymes of rat brain. , 1985, Toxicology letters.

[24]  W. Stolz,et al.  A randomized two‐sided placebo‐controlled study on the efficacy and safety of atmospheric non‐thermal argon plasma for pruritus , 2013, Journal of the European Academy of Dermatology and Venereology : JEADV.

[25]  U. Jakob,et al.  Thiol-based redox switches in eukaryotic proteins. , 2009, Antioxidants & redox signaling.

[26]  S. McMahon,et al.  Cold atmospheric pressure plasma jet interactions with plasmid DNA , 2011 .

[27]  E. Takai,et al.  Protein Inactivation by Low‐temperature Atmospheric Pressure Plasma in Aqueous Solution , 2012 .

[28]  Mattia Adamo,et al.  Glutathionylation of cytosolic glyceraldehyde-3-phosphate dehydrogenase from the model plant Arabidopsis thaliana is reversed by both glutaredoxins and thioredoxins in vitro. , 2012, The Biochemical journal.

[29]  J. Wood,et al.  In vivo fluorescence excitation spectroscopy and in vivo Fourier‐transform Raman spectroscopy in human skin: evidence of H2O2 oxidation of epidermal albumin in patients with vitiligo , 2004 .

[30]  Howard C. Berg,et al.  Genetic analysis , 1957, Nature Biotechnology.

[31]  G. Georghiou,et al.  Bactericidal Action of the Reactive Species Produced by Gas-Discharge Nonthermal Plasma at Atmospheric Pressure: A Review , 2006, IEEE Transactions on Plasma Science.

[32]  Seong-Mi Kim,et al.  Decomposition of biological macromolecules by plasma generated with helium and oxygen. , 2006, Journal of microbiology.

[33]  DNA damage in mammalian cells by non-thermal atmospheric pressure microsecond pulsed dielectric barrier discharge plasma is not mediated by ozone , 2010, ICOPS 2010.

[34]  A. le Pape,et al.  Effects of a Non Thermal Plasma Treatment Alone or in Combination with Gemcitabine in a MIA PaCa2-luc Orthotopic Pancreatic Carcinoma Model , 2012, PloS one.

[35]  W H Glaze,et al.  Reaction products of ozone: a review. , 1986, Environmental health perspectives.

[36]  G. Thomas,et al.  Characterization of DNA structures by laser Raman spectroscopy , 1984, Biopolymers.

[37]  Robert C. Wolpert,et al.  A Review of the , 1985 .

[38]  Norman M. Dott.  An Introductory Review , 1962 .

[39]  M. Schön,et al.  Atmospheric pressure plasma in dermatology: Ulcus treatment and much more , 2013 .

[40]  A. Garner,et al.  Nonthermal Atmospheric Plasma Rapidly Disinfects Multidrug-Resistant Microbes by Inducing Cell Surface Damage , 2012, Antimicrobial Agents and Chemotherapy.

[41]  J. Alonso,et al.  Double-strand break repair in bacteria: a view from Bacillus subtilis. , 2011, FEMS microbiology reviews.

[42]  Wilhelm Stolz,et al.  Cold atmospheric plasma: a successful treatment of lesions in Hailey-Hailey disease. , 2011, Archives of dermatology.

[43]  J. Zimmermann,et al.  Contact-Free Inactivation of Candida albicans Biofilms by Cold Atmospheric Air Plasma , 2012, Applied and Environmental Microbiology.

[44]  J. Benedikt,et al.  Inactivation of Bacillus atrophaeus and of Aspergillus niger using beams of argon ions, of oxygen molecules and of oxygen atoms , 2008 .

[45]  J. Lackmann,et al.  The Role of VUV Radiation in the Inactivation of Bacteria with an Atmospheric Pressure Plasma Jet , 2011, 1105.6260.

[46]  M Landthaler,et al.  A first prospective randomized controlled trial to decrease bacterial load using cold atmospheric argon plasma on chronic wounds in patients , 2010, The British journal of dermatology.

[47]  J. Lackmann,et al.  Separation of VUV/UV photons and reactive particles in the effluent of a He/O2 atmospheric pressure plasma jet , 2011, 1105.2207.

[48]  J. Lackmann,et al.  Characterization of Damage to Bacteria and Bio-macromolecules Caused by (V)UV Radiation and Particles Generated by a Microscale Atmospheric Pressure Plasma Jet , 2012 .

[49]  Gregor E. Morfill,et al.  Plasma medicine: an introductory review , 2009 .

[50]  H. Sahl,et al.  Proteomic Response of Bacillus subtilis to Lantibiotics Reflects Differences in Interaction with the Cytoplasmic Membrane , 2012, Antimicrobial Agents and Chemotherapy.