INFLUENCE OF CARBOPOL GEL WITH MELANINE ON CONTENT OF CYTOKINS IN THE WOUNDS SUBSTRATE OF FULL-THICKNESS EXCISIONAL WOUNDS OF SKIN IN THE RATS

Nikitina N. S., Stepanova L. I., Vereschaka V. V., Savchuk O. M., Beregova T. V

CLINICAL AND EXPERIMENTAL MEDICINE

Scentific article

Formerly we have shown that the use carbopol gel with melanin accelerates wound healing without the formation of rough scar. However, the mechanism of dermatotropic action of melanin is not studied until the end. The purpose of the study was to determine the content of cytokines in wound substrates of full-thickness excisional wounds of skin in the rats after their treatment with gel of carbopol with melanin. Object and methods. The research was conducted on 80 white laboratory rats weighing 200-220 g which were kept in the animals house of the Educational and Scientific Center “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv. Rats were divided into three groups: I – the intact control group, II – the control group with wounds without treatment, III, IV – experimental groups in which the wounds were treated with 0,5% gel of carbopol and 0,5% gel of carbopol with melanin (0,1%) consequently. The modeling of full-thickness excisional wounds of skin in the rats was carried out in accordance with the Berchenko G.N. On the 9th day of experiment and in the day of complete wound closure in wound substrate the content of interleukin-1b (IL-1b), IL-6 and IL-10 were assayed by ELISA according to the manufacturer’s recommendations (R&D Systems). Skin samples were immobilized onto 96-well plate and incubated with corresponding specific primary antibodies (Santa Cruz, USA). After that secondary antibodies conjugated with horseradish peroxidase (Bio- Rad, USA) were added. To enable colorimetric detection, reaction with the substrate o’phenylenediamine/hydrogen peroxide (Sigma, USA) was performed and absorbance of each well was read at 422 nm (“Synergy”, BioTek, USA). Values were expressed as optical density / mg of protein. Total proteins were determined by Bradford’s method. Results. It was established that in rats with wounds without treatment on the 9th day of study in the wound substrate the content of IL-1b increased by 29,0% (p<0,01), the content of IL-6 increased by 34,2% (p<0,01) and the content of IL-10 increased by 60,0% (р<0,001). On the 9th day of study in rats after wounds treatment with 0,5% gel of carbopol the content of IL-1b, IL-6 and IL-10 was the same as in the control group. Under the conditions of daily treatment of wounds with 0.5% gel of carbopol with melanin (0.1%), the content of and the cytokines in the wound substrate on the 9th day after wound modeling was at the level of intact control. On the day of complete wound closure, the content in the wound substrate of the studied cytokines was at the level of intact control.

melanin, full-thickness excisional wounds, wounds substrate, interleukin-1b, interleukin-6, interleukin-10

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«Bulletin of problems biology and medicine» Issue 2 Part 1 (150), 2019 year, 168-171 pages, index UDK 616-001.4-085.33:615.03.032:612-092.9

10.29254/2077-4214-2019-2-1-150-168-171