[en] Cold plasmas have found their application in a wide range of biomedical fields by virtue of their high chemical reactivity. In the past decades, many attempts have been made to use cold plasmas in wound healing, and within this field, many studies have focused on plasma-induced cell proliferation mechanisms. In this work, one step further has been taken to demonstrate the advanced role of plasma in wound healing. To this end, the simultaneous ability of plasma to induce cell proliferation and permeabilize treated cells has been examined in the current study. The driving force was to advance the wound healing effect of plasma with drug delivery. On this subject, we demonstrate in vitro the healing effect of Ar, Ar+N2 plasma, and their aerosol counterparts. A systematic study has been carried out to study the role of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in cell adhesion, signaling, differentiation, and proliferation. An additional investigation was also performed to study the permeabilization of cells and the delivery of the modeled drug carrier fluorescein isothiocyanate (FITC) labeled dextran into cells upon plasma treatment. Short 35 s plasma treatments were found to promote fibroblast adhesion, migration, signaling, proliferation, and differentiation by means of reactive oxygen and nitrogen species (RONS) created by plasma and deposited into the cell environment. The impact of the plasma downstream products NO2- and NO3- on the expressions of the focal adhesion's genes, syndecans, and collagens was observed to be prominent. On the other hand, the differentiation of fibroblasts to myofibroblasts was mainly initiated by ROS produced by the plasma. In addition, the ability of plasma to locally permeabilize fibroblast cells was demonstrated. During proliferative cell treatment, plasma can simultaneously induce cell membrane permeabilization (d ∼ 7.3 nm) by the species OH and H2O2. The choice for a plasma or a plasma-aerosol configuration thus allows the possibility to change the spatial chemistry of drug delivery molecules and thus to locally deliver drugs. Accordingly, this study offers a pivotal step toward plasma-assisted wound healing advanced by drug delivery.
Disciplines :
Engineering, computing & technology: Multidisciplinary, general & others
Author, co-author :
Sremački, Ivana ; Department of Applied Physics, Ghent University, Sint-Pietersnieuwstraat 41, Gent 9000, Belgium
Asadian, Mahtab ; Department of Applied Physics, Ghent University, Sint-Pietersnieuwstraat 41, Gent 9000, Belgium ; Skeletal Biology & Engineering Research Center, ON1 Herestraat 49, 3000 Leuven, Belgium
De Geyter, Nathalie; Department of Applied Physics, Ghent University, Sint-Pietersnieuwstraat 41, Gent 9000, Belgium
Leys, Christophe; Department of Applied Physics, Ghent University, Sint-Pietersnieuwstraat 41, Gent 9000, Belgium
Geris, Liesbet ; Université de Liège - ULiège > Département d'aérospatiale et mécanique > Génie biomécanique ; Skeletal Biology & Engineering Research Center, ON1 Herestraat 49, 3000 Leuven, Belgium
Nikiforov, Anton ; Department of Applied Physics, Ghent University, Sint-Pietersnieuwstraat 41, Gent 9000, Belgium
Language :
English
Title :
Potentials of a Plasma-Aerosol System for Wound Healing Advanced by Drug Introduction: An In Vitro Study.
This study was performed as a collaboration between Ghent University and KU Leuven and was supported by the FWO project “Plasma-skin interactions: from wound treatment to topical introduction of molecules”, number G084917N.
Singer, A. J.; Clark, R. A. Cutaneous wound healing. New England journal of medicine 1999, 341 ( 10), 738- 746, 10.1056/NEJM199909023411006
Vaughan, M. B.; Howard, E. W.; Tomasek, J. J. Transforming growth factor-β1 promotes the morphological and functional differentiation of the myofibroblast. Experimental cell research 2000, 257 ( 1), 180- 189, 10.1006/excr.2000.4869
Smet, S.; Probst, S.; Holloway, S.; Fourie, A.; Beele, H.; Beeckman, D. The measurement properties of assessment tools for chronic wounds: a systematic review. International Journal of Nursing Studies 2021, 121, 103998, 10.1016/j.ijnurstu.2021.103998
Kessler, L.; Bilbault, P.; ORTega, F.; Grasso, C.; Passemard, R.; Stephan, D.; Pinget, M.; Schneider, F. Hyperbaric oxygenation accelerates the healing rate of nonischemic chronic diabetic foot ulcers: a prospective randomized study. Diabetes care 2003, 26 ( 8), 2378- 2382, 10.2337/diacare.26.8.2378
Moffett, J.; Griffin, N. E.; Ritz, M. C.; George, F. R. Pulsed radio frequency energy field treatment of cells in culture results in increased expression of genes involved in the inflammation phase of lower extremity diabetic wound healing. Journal of Diabetic Foot Complications 2010, 2 ( 3), 57- 64
Eaglstein, W. H.; Falanga, V. Tissue engineering and the development of Apligraf®, a human skin equivalent. Clinical therapeutics 1997, 19 ( 5), 894- 905, 10.1016/S0149-2918(97)80043-4
Kiwanuka, E.; Junker, J.; Eriksson, E. Harnessing growth factors to influence wound healing. Clinics in plastic surgery 2012, 39 ( 3), 239- 248, 10.1016/j.cps.2012.04.003
Frykberg, R. G.; Banks, J. Challenges in the treatment of chronic wounds. Advances in wound care 2015, 4 ( 9), 560- 582, 10.1089/wound.2015.0635
Ferguson, M.; Byrnes, C.; Sun, L.; Marti, G.; Bonde, P.; Duncan, M.; Harmon, J. W. Wound healing enhancement: electroporation to address a classic problem of military medicine. World journal of surgery 2005, 29, S55- S59, 10.1007/s00268-004-2062-2
Sun, L.; Xu, L.; Chang, H.; Henry, F. A.; Miller, R. M.; Harmon, J. M.; Nielsen, T. B. Transfection with aFGF cDNA improves wound healing. Journal of investigative dermatology 1997, 108 ( 3), 313- 318, 10.1111/1523-1747.ep12286471
Sen, C. K. The general case for redox control of wound repair. Wound repair and regeneration 2003, 11 ( 6), 431- 438, 10.1046/j.1524-475X.2003.11607.x
Sen, C. K.; Khanna, S.; Babior, B. M.; Hunt, T. K.; Ellison, E. C.; Roy, S. Oxidant-induced vascular endothelial growth factor expression in human keratinocytes and cutaneous wound healing. J. Biol. Chem. 2002, 277 ( 36), 33284- 33290, 10.1074/jbc.M203391200
Sen, C. K. Wound healing essentials: let there be oxygen. Wound repair and regeneration 2009, 17 ( 1), 1- 18, 10.1111/j.1524-475X.2008.00436.x
Fridman, G.; Friedman, G.; Gutsol, A.; Shekhter, A. B.; Vasilets, V. N.; Fridman, A. Applied plasma medicine. Plasma processes and polymers 2008, 5 ( 6), 503- 533, 10.1002/ppap.200700154
Bekeschus, S.; von Woedtke, T.; Emmert, S.; Schmidt, A. Medical gas plasma-stimulated wound healing: Evidence and mechanisms. Redox Biology 2021, 46, 102116, 10.1016/j.redox.2021.102116
Privat-Maldonado, A.; Schmidt, A.; Lin, A.; Weltmann, K.-D.; Wende, K.; Bogaerts, A.; Bekeschus, S. ROS from physical plasmas: Redox chemistry for biomedical therapy. Oxidative Medicine and Cellular Longevity 2019, 2019, 9062098, 10.1155/2019/9062098
Kalghatgi, S.; Friedman, G.; Fridman, A.; Clyne, A. M. Endothelial cell proliferation is enhanced by low dose non-thermal plasma through fibroblast growth factor-2 release. Annals of biomedical engineering 2010, 38 ( 3), 748- 757, 10.1007/s10439-009-9868-x
Kamata, H.; Hirata, H. Redox regulation of cellular signalling. Cellular signalling 1999, 11 ( 1), 1- 14, 10.1016/S0898-6568(98)00037-0
Fridman, G.; Peddinghaus, M.; Fridman, A.; Balasubramanian, M.; Gutsol, A.; Friedman, G. Use of non-thermal atmospheric pressure plasma discharge for coagulation and sterilization of surface wounds. In 32nd IEEE International Conference on Plasma Science, 2005; p 257.
Mohd Nasir, N.; Lee, B.K.; Yap, S.S.; Thong, K.L.; Yap, S.L. Cold plasma inactivation of chronic wound bacteria. Archives of biochemistry and biophysics 2016, 605, 76- 85, 10.1016/j.abb.2016.03.033
Laroussi, M. Low temperature plasma-based sterilization: overview and state-of-the-art. Plasma Process. Polym. 2005, 2 ( 5), 391- 400, 10.1002/ppap.200400078
Kalghatgi, S. U.; Fridman, A.; Friedman, G.; Clyne, A. M. Cell proliferation following non-thermal plasma is related to reactive oxygen species induced fibroblast growth factor-2 release. In 2009 Annual International Conference of the IEEE Engineering in Medicine and Biology Society; IEEE, 2009; pp 6030- 6033.
Park, J.; Lee, H.; Lee, H. J.; Kim, G. C.; Kim, S.-S.; Han, S.; Song, K. Non-thermal atmospheric pressure plasma is an excellent tool to activate proliferation in various mesoderm-derived human adult stem cells. Free Radical Biol. Med. 2019, 134, 374- 384, 10.1016/j.freeradbiomed.2019.01.032
Laroussi, M. The Resistive Barrier Discharge: A Brief Review of the Device and Its Biomedical Applications. Plasma 2021, 4 ( 1), 75- 80, 10.3390/plasma4010004
Breathnach, R.; McDonnell, K. A.; Chebbi, A.; Callanan, J. J.; Dowling, D. P. Evaluation of the effectiveness of kINPen Med plasma jet and bioactive agent therapy in a rat model of wound healing. Biointerphases 2018, 13 ( 5), 051002, 10.1116/1.5046489
Naik, A.; Kalia, Y. N.; Guy, R. H. Transdermal drug delivery: overcoming the skin’s barrier function. Pharmaceutical science & technology today 2000, 3 ( 9), 318- 326, 10.1016/S1461-5347(00)00295-9
Schoellhammer, C. M.; Blankschtein, D.; Langer, R. Skin permeabilization for transdermal drug delivery: recent advances and future prospects. Expert opinion on drug delivery 2014, 11 ( 3), 393- 407, 10.1517/17425247.2014.875528
Vanbever, R.; Preat, V. In vivo efficacy and safety of skin electroporation. Advanced drug delivery reviews 1999, 35 ( 1), 77- 88, 10.1016/S0169-409X(98)00064-7
Sersa, G.; Miklavcic, D.; Cemazar, M.; Rudolf, Z.; Pucihar, G.; Snoj, M. Electrochemotherapy in treatment of tumours. European Journal of Surgical Oncology (EJSO) 2008, 34 ( 2), 232- 240, 10.1016/j.ejso.2007.05.016
Van Meirvenne, S.; Straetman, L.; Heirman, C.; Dullaers, M.; De Greef, C.; Van Tendeloo, V.; Thielemans, K. Efficient genetic modification of murine dendritic cells by electroporation with mRNA. Cancer gene therapy 2002, 9 ( 9), 787- 797, 10.1038/sj.cgt.7700499
Sremački, I.; Jurov, A.; Modic, M.; Cvelbar, U.; Wang, L.; Leys, C.; Nikiforov, A. Y. On diagnostics of an annular shape RF plasma jet operating in Ar at atmospheric conditions. Plasma Sources Science and Technology 2020, 29, 035027, 10.1088/1361-6595/ab71f7
Sremački, I.; Bruno, G.; Jablonowski, H.; Leys, C.; Nikiforov, A. Y.; Wende, K. Influence of aerosol injection on the liquid chemistry induced by an RF argon plasma jet. Plasma Sources Science and Technology 2021, 30, 095018, 10.1088/1361-6595/abe176
Sremački, I.; Kos, S. p.; Bošnjak, M. a.; Jurov, A.; Serša, G.; Modic, M.; Leys, C.; Cvelbar, U.; Nikiforov, A. Plasma Damage Control: From Biomolecules to Cells and Skin. ACS Appl. Mater. Interfaces 2021, 13 ( 39), 46303- 46316, 10.1021/acsami.1c12232
Labay, C.; Roldán, M.; Tampieri, F.; Stancampiano, A.; Bocanegra, P. E.; Ginebra, M.-P.; Canal, C. Enhanced generation of reactive species by cold plasma in gelatin solutions for selective cancer cell death. ACS Appl. Mater. Interfaces 2020, 12 ( 42), 47256- 47269, 10.1021/acsami.0c12930
Park, J. H.; Kim, M.; Shiratani, M.; Cho, A. E.; Choi, E. H.; Attri, P. Variation in structure of proteins by adjusting reactive oxygen and nitrogen species generated from dielectric barrier discharge jet. Sci. Rep. 2016, 6 ( 1), 1- 14, 10.1038/srep35883
Van Gaens, W.; Bogaerts, A. Kinetic modelling for an atmospheric pressure argon plasma jet in humid air. J. Phys. D: Appl. Phys. 2013, 46 ( 27), 275201, 10.1088/0022-3727/46/27/275201
Ngo Thi, M. H.; Shao, P. L.; Liao, J. D.; Lin, C. C. K.; Yip, H. K. Enhancement of angiogenesis and epithelialization processes in mice with burn wounds through ROS/RNS signals generated by non-thermal N2/Ar micro-plasma. Plasma Process. Polym. 2014, 11 ( 11), 1076- 1088, 10.1002/ppap.201400072
Livak, K. J.; Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2- ΔΔCT method. methods 2001, 25 ( 4), 402- 408, 10.1006/meth.2001.1262
Leduc, M.; Guay, D.; Leask, R.; Coulombe, S. Cell permeabilization using a non-thermal plasma. New J. Phys. 2009, 11 ( 11), 115021, 10.1088/1367-2630/11/11/115021
Davies, K. J. The broad spectrum of responses to oxidants in proliferating cells: a new paradigm for oxidative stress. IUBMB life 1999, 48 ( 1), 41- 47, 10.1080/152165499307404
Dobrynin, D.; Wasko, K.; Friedman, G.; Fridman, A. A.; Fridman, G. Cold plasma sterilization of open wounds: live rat model. Plasma Medicine 2011, 1 ( 2), 109, 10.1615/PlasmaMed.2011002698
Lee, J.; Bong, C.; Lim, W.; Bae, P. K.; Abafogi, A. T.; Baek, S. H.; Shin, Y.-B.; Bak, M. S.; Park, S. Fast and easy disinfection of coronavirus-contaminated face masks using ozone gas produced by a dielectric barrier discharge plasma generator. Environ. Sci. Technol. Lett. 2021, 8 ( 4), 339- 344, 10.1021/acs.estlett.1c00089
Boncler, M.; Różalski, M.; Krajewska, U.; Podsędek, A.; Watala, C. Comparison of PrestoBlue and MTT assays of cellular viability in the assessment of anti-proliferative effects of plant extracts on human endothelial cells. Journal of pharmacological and toxicological methods 2014, 69 ( 1), 9- 16, 10.1016/j.vascn.2013.09.003
Smith, M.; Barbenel, J.; Courtney, J.; Grant, M. Novel quantitative methods for the determination of biomaterial cytotoxicity; SAGE Publications Sage UK: London, England, 1992.
Yonson, S.; Coulombe, S.; Leveille, V.; Leask, R. Cell treatment and surface functionalization using a miniature atmospheric pressure glow discharge plasma torch. J. Phys. D: Appl. Phys. 2006, 39 ( 16), 3508, 10.1088/0022-3727/39/16/S08
Han, G.; Ceilley, R. Chronic wound healing: a review of current management and treatments. Advances in therapy 2017, 34 ( 3), 599- 610, 10.1007/s12325-017-0478-y
Darby, I. A.; Hewitson, T. D. Fibroblast differentiation in wound healing and fibrosis. International review of cytology 2007, 257, 143- 179, 10.1016/S0074-7696(07)57004-X
Schmidt, A.; Liebelt, G.; Nießner, F.; von Woedtke, T.; Bekeschus, S. Gas plasma-spurred wound healing is accompanied by regulation of focal adhesion, matrix remodeling, and tissue oxygenation. Redox biology 2021, 38, 101809, 10.1016/j.redox.2020.101809
Afratis, N. A.; Nikitovic, D.; Multhaupt, H. A.; Theocharis, A. D.; Couchman, J. R.; Karamanos, N. K. Syndecans-key regulators of cell signaling and biological functions. FEBS journal 2017, 284 ( 1), 27- 41, 10.1111/febs.13940
Lukes, P.; Dolezalova, E.; Sisrova, I.; Clupek, M. Aqueous-phase chemistry and bactericidal effects from an air discharge plasma in contact with water: evidence for the formation of peroxynitrite through a pseudo-second-order post-discharge reaction of H2O2 and HNO2. Plasma Sources Science and Technology 2014, 23 ( 1), 015019, 10.1088/0963-0252/23/1/015019
Haertel, B.; Von Woedtke, T.; Weltmann, K.-D.; Lindequist, U. Non-thermal atmospheric-pressure plasma possible application in wound healing. Biomolecules & therapeutics 2014, 22 ( 6), 477, 10.4062/biomolther.2014.105
Barrientos, S.; Brem, H.; Stojadinovic, O.; Tomic-Canic, M. Clinical application of growth factors and cytokines in wound healing. Wound Repair and Regeneration 2014, 22 ( 5), 569- 578, 10.1111/wrr.12205
Huang, S.; Huang, G. Preparation and drug delivery of dextran-drug complex. Drug delivery 2019, 26 ( 1), 252- 261, 10.1080/10717544.2019.1580322
Dhaneshwar, S.; Bhilare, N.; Roy, S. Dextran Pharmaceutical Applications. In Polysaccharides of Microbial Origin: Biomedical Applications; Springer, 2021; pp 1- 28.
Ludtke, J. J.; Sebestyén, M. G.; Wolff, J. A. The effect of cell division on the cellular dynamics of microinjected DNA and dextran. Molecular Therapy 2002, 5 ( 5), 579- 588, 10.1006/mthe.2002.0581
Larina, I. V.; Evers, B. M.; Esenaliev, R. O. Optimal drug and gene delivery in cancer cells by ultrasound-induced cavitation. Anticancer research 2005, 25 ( 1A), 149- 156
Guignet, E. G.; Meyer, T. Suspended-drop electroporation for high-throughput delivery of biomolecules into cells. Nat. Methods 2008, 5 ( 5), 393- 395, 10.1038/nmeth.1201
Stoffels, E.; Sakiyama, Y.; Graves, D. B. Cold atmospheric plasma: charged species and their interactions with cells and tissues. IEEE Trans. Plasma Sci. 2008, 36 ( 4), 1441- 1457, 10.1109/TPS.2008.2001084
Dobrynin, D.; Fridman, G.; Friedman, G.; Fridman, A. Physical and biological mechanisms of direct plasma interaction with living tissue. New J. Phys. 2009, 11 ( 11), 115020, 10.1088/1367-2630/11/11/115020
Oinuma, G.; Nayak, G.; Du, Y.; Bruggeman, P. J. Controlled plasma-droplet interactions: A quantitative study of OH transfer in plasma-liquid interaction. Plasma Sources Science and Technology 2020, 29 ( 9), 095002, 10.1088/1361-6595/aba988
Bruggeman, P. J.; Kushner, M. J.; Locke, B. R.; Gardeniers, J. G.; Graham, W.; Graves, D. B.; Hofman-Caris, R.; Maric, D.; Reid, J. P.; Ceriani, E. Plasma-liquid interactions: a review and roadmap. Plasma sources science and technology 2016, 25 ( 5), 053002, 10.1088/0963-0252/25/5/053002
Burlica, R.; Shih, K.-Y.; Locke, B. Formation of H2 and H2O2 in a water-spray gliding arc nonthermal plasma reactor. Ind. Eng. Chem. Res. 2010, 49 ( 14), 6342- 6349, 10.1021/ie100038g
Rybenkov, V. V.; Vologodskii, A. V.; Cozzarelli, N. R. The effect of ionic conditions on the conformations of supercoiled DNA. I. Sedimentation analysis. Journal of molecular biology 1997, 267 ( 2), 299- 311, 10.1006/jmbi.1996.0876
Sakai, Y.; Khajoee, V.; Ogawa, Y.; Kusuhara, K.; Katayama, Y.; Hara, T. A novel transfection method for mammalian cells using gas plasma. Journal of biotechnology 2006, 121 ( 3), 299- 308, 10.1016/j.jbiotec.2005.08.020