Authors
Markevich P.S.1, Filippov A.V.1, Dolgikh R.N.1, Borovkova N.V.2, 3, 4, Makarov M.S.2, Storozheva M.V.2, Ponomarev I.N.2, Zelenina T.A.5, Zemlyanoy A.B.1
1 National medical state center of surgery named after A.V. Vishnevsky, Moscow
2 N.V. Sklifosovsky Research Institute for Emergency Medicine, Moscow
3 N.I. Pirogov Russian National Research Medical University, Moscow
4 Russian Medical Academy of Continuing Professional Education, Moscow
5 S.M. Kirov Military medical academy, St. Petersburg
Abstract
Background: Controlled generation of active forms of oxygen and nitrogen stimulates wound healing processes. One of the sources of controlled generation of active forms of oxygen and nitrogen is low-temperature argon plasma. Low-temperature plasma activates various forms of cellular activity. However, the subtle mechanisms of such stimulation remain unclear. Thus, there is a need for a detailed study of the biological effect of low-temperature argon plasma on human cells.
Objective: to evaluate the effect of low-temperature argon plasma on the structure and proliferative activity of human fibroblasts in vitro.
Materials and methods. Human fibroblast culture (line M-21, passage 25, Chumakov Institute of Poliomyelitis and Viral Encephalitis) was used in the work. After exposure to low-temperature argon plasma, the cells were cultured for 3 days in DMEM/F12 medium supplemented with 10% Gibco bovine embryo serum, then morphofunctional analysis of the cells was performed using vital staining. After 3 days, the cells were passaged (reseeded in other Petri dishes with a 1:2 dilution), then the cells were cultured for 3 days, and a repeated morphofunctional analysis was performed. All work with cell cultures was carried out in accordance with the international GMP standard.
Results. Stimulation of the proliferative activity of fibroblasts was noted in the second passage after treatment with low-temperature argon plasma from a distance of 10 cm for 30 and 45 seconds and 15 cm for 45 seconds.The use of a collagen bandage screen significantly increases cell survival when exposed to plasma from a distance of 10 cm for 15 seconds. With screening, the decrease in the integrity of cell membranes after plasma exposure is less pronounced than with exposure without screening. When reseeding, cells treated in the presence of a collagen screen exhibit high proliferative activity and restore membrane integrity.
Keywords: low-temperature argon plasma, fibroblasts, proliferation.
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