1 Scientific RepoRts | 6:19144 | DOI: 10.1038/srep19144 www.nature.com/scientificreports Investigation on the efects of the atmospheric pressure plasma on wound healing in diabetic rats sara Fathollah 1,* , shahriar Mirpour 2,* , parvin Mansouri 3 , Ahmad Reza Dehpour 4,5 , Mahmood Ghoranneviss 1 , Nastaran Rahimi 4,5 , Zahra Safaie Naraghi 6 , Reza Chalangari 7 & Katalin Martits Chalangari 7 It is estimated that 15 percent of individuals with diabetes mellitus sufer from diabetic ulcers worldwide. The aim of this study is to present a non-thermal atmospheric plasma treatment as a novel therapy for diabetic wounds. The plasma consists of ionized helium gas that is produced by a high-voltage (8 kV) and high-frequency (6 kHz) power supply. Diabetes was induced in rats via an intravascular injection of streptozotocin. The plasma was then introduced to artiicial xerograph wounds in the rats for 10 minutes. Immunohistochemistry assays was performed to determine the level of transforming growth factor (TGF-β1) cytokine. The results showed a low healing rate in the diabetic wounds compared with the wound-healing rate in non-diabetic animals (P < 0.05). Moreover, the results noted that plasma enhanced the wound-healing rate in the non-diabetic rats (P < 0.05), and signiicant wound contraction occurred after the plasma treatment compared with untreated diabetic wounds (P < 0.05). Histological analyses revealed the formation of an epidermis layer, neovascularization and cell proliferation. The plasma treatment also resulted in the release of TGF-β1 cytokine from cells in the tissue medium. The indings of this study demonstrate the efect of plasma treatment for wound healing in diabetic rats. Wound healing is a common concern in diabetic patients. Patients with diabetes have wound healing complica- tions associated with many factors, including neuropathy, vascular disease, and foot deformities 1 . he conven- tional therapeutic strategies are limited to wound dressings, cell therapy, and oxygen therapy. hese methods are not completely successful in curing diabetic wounds because of the slow wound healing process, the high cost of therapies, and the lack of improvement in the disturbed healing process caused by diabetes 2 . Non-thermal atmospheric pressure plasma, which has been published as a possible new treatment in chronic wound therapy, may hopefully solve this issue. Plasmas are partially-ionized gases and are described as the fourth state of matter. he word “plasma” was coined by Irving Langmuir, for the ionized gas which is comprised of suspended electrons, ions, and other excited and charged particles 3,4 . Medical applications of non-thermal plasma are classiied into direct plasma and indirect plasma therapy; the former is used directly on the human body, while the later is generated remotely and then transported to the surface to be treated 5 . Plasma application creates a new ield (at the precinct) between plasma science and biomedicine. In recent years, atmospheric pressure plasma has demonstrated many potential applica- tions in skin wound healing 6 , blood coagulation 7,8 , bacteria disinfection 9,10 and cancer therapy 11,12 . he possibility of potentiation of wound healing by atmospheric plasma has been the focus of attention in many recent stud- ies 13–15 . Plasma can produce reactive species such as Nitric oxide (NO), Hydroxyl (OH) and Atomic oxygen (O) which is an important reaction by living microorganisms 16 . his special tissue-reactive species interaction may lead to the acceleration of tissue repair processes without adverse efects on normal tissue thereby diferentiating 1 Plasma Physics Research center, Science and Research branch of islamic Azad University, tehran, iran. 2 Laser and Plasma institute, Shahid Beheshti University, tehran, iran. 3 Skin and Stem cell Research center, tehran University of Medical Sciences, tehran, iran. 4 experimental Medicine Research center, tehran University of Medical Science, tehran, iran. 5 Department of Pharmacology, School of Medicine, tehran University of Medical Sciences, tehran, iran. 6 Department of Pathology, Razi Skin Hospital, tehran University of Medical Sciences, tehran, iran. 7 Kassir Dermatology, 8335 Walnut Hill Ln, #140, Dallas, TX 75231, USA. * these authors contributed equally to this work. correspondence and requests for materials should be addressed to P.M. (email: mansorip@sina.tums.ac.ir) Received: 27 September 2015 Accepted: 07 December 2015 Published: 23 February 2016 opeN