The Effectiveness of Local Application of Melatonin in the Original Dermal Film in Experimental Thermal Trauma

ORIGINAL ARTICLE
M. V. Osikov, A. A. Ageeva, Yu. I. Ageev, A. A. Fedosov, K. V. Nikushkina, Yu. V. Loginova

 
International Journal of Biomedicine. 2021;11(4):581-589.
DOI: 10.21103/Article11(4)_OA30
Originally published December 10, 2021

Abstract: 

Background: The development and pathogenetic substantiation of the new agents used for local therapy of thermal trauma (TT) is an urgent problem in medicine. Melatonin (MT) is an endogenous factor of homeostasis regulation with pleiotropic potential. The aim of our study was to assess the morphology, expression of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF), indicators of repair, oxidative destruction of lipids in the skin lesion focus in the dynamics of experimental TT under the conditions of using the original dermal film (DF) with MT.
Methods and Results: The experiment was performed on 104 male Wistar rats weighing 200-240 g. For modeling TT II degree according to ICD-10 a relative area of 3.5% of the body surface, an interscapular region isolated from the surrounding tissues, was immersed in distilled water at 98-99 °C for 12 sec. DF based on sodium carboxymethylcellulose with an area of 12 cm2 with MT at a concentration of 5 mg/g was applied daily for five days. The wound area and epithelialization rate were calculated. The content of MMP-9 and VEGF in the burn wound was assessed by an immunohistochemical method.  In the homogenate of the burn wound, the content of LPO products was assessed. Morphological and biochemical studies were performed on Days 5, 10 and 20 after TT induction.
With experimental TT from Day 5 to Day 20, the absolute area of ​​the burn wound decreases by 35%, the rate of epithelialization increases, the number of neutrophils in the focus of thermal damage decreases, while the representation of lymphocytes, histiocytes, and fibroblasts increases; the expression of MMP-9 and VEGF increases; predominantly secondary and final LPO products in the heptane phase accumulate, the final products of LPO in the isopropanol phase of the lipid extract. The use of MT in the composition of DF daily for 5 days with experimental TT leads to a decrease in the area of ​​the wound defect (by 46% of the original area on Day 20), an increase in the rate of its epithelialization, an increase in the content of lymphocytes and fibroblasts in the burn wound on Days 5, 10 and 20 of TT, a decrease in the representation of neutrophils and macrophages on Days 5 and 10, as well as an increase in VEGF expression on Days 5 and 10, MMP-9 - on Day 5 and a decrease in MMP-9 expression on Days 10 and 20 of TT. In addition, the use of MT in the composition of DF leads to a decrease in the content of predominantly secondary and end products of LPO in the heptane and isopropanol phases of the burn wound on Days 10 and 20 of TT. Correlation analysis revealed that a decrease in the burn surface area under a local application of MT occurs with an increase in the content of VEGF in the wound area and a decrease in the content of MMP-9 and secondary and final LPO products in the heptane phase and the isopropanol phase. On Day 20, there were direct moderate correlations between the absolute burn surface area, on one hand, and secondary and final LPO products, on the other, in the heptane phase (R=0.51, R=0,68; P<0.05) and the isopropanol phase (R=0.44, R=0.46; P<0.05), respectively.
Conclusion: The results obtained expand the existing understanding of the role of changes in the expression of MMP-9 and VEGF in the pathogenesis of TT. We believe that the repair-stimulating effect of MT in the DF, which we established during TT at the preclinical stage, is associated with the LPO-limiting effect of MT and a change in the expression of MMP-9 and VEGF in the burn wound and is a prerequisite for further study of the mechanism of action and the effectiveness of MT application in clinical conditions in TT.

Keywords: 
MMP-9 • VEGF • lipid peroxidation • thermal trauma • melatonin
References: 
  1. Hirche C, Citterio A, Hoeksema H, Koller J, Lehner M, Martinez JR, Monstrey S, Murray A, Plock JA, Sander F, Schulz A, Ziegler B, Kneser U. Eschar removal by bromelain based enzymatic debridement (Nexobrid®) in burns: An European consensus. Burns. 2017 Dec;43(8):1640-1653. doi: 10.1016/j.burns.2017.07.025. 
  2. Serebrakian AT, Pickrell BB, Varon DE, Mohamadi A, Grinstaff MW, Rodriguez EK, Nazarian A, Halvorson EG, Sinha I. Meta-analysis and Systematic Review of Skin Graft Donor-site Dressings with Future Guidelines. Plast Reconstr Surg Glob Open. 2018 Sep 24;6(9):e1928. doi: 10.1097/GOX.0000000000001928. 
  3. Kim Y, Kym D, Cho YS, Yoon J, Yim H, Hur J, Chun W. Use of Fibrin Sealant for Split-Thickness Skin Grafts in Patients with Hand Burns: A Prospective Cohort Study. Adv Skin Wound Care. 2018 Dec;31(12):551-555. doi: 10.1097/01.ASW.0000547413.61758.27.
  4. Huang J, Ren J, Chen G, Li Z, Liu Y, Wang G, Wu X. Tunable sequential drug delivery system based on chitosan/hyaluronic acid hydrogels and PLGA microspheres for management of non-healing infected wounds. Mater Sci Eng C Mater Biol Appl. 2018 Aug 1;89:213-222. doi: 10.1016/j.msec.2018.04.009. 
  5. Mullens CL, Messa CA 4th, Kozak GM, Rhemtulla IA, Fischer JP. To Glue or Not to Glue? Analysis of Fibrin Glue for Split-thickness Skin Graft Fixation. Plast Reconstr Surg Glob Open. 2019 May 16;7(5):e2187. doi: 10.1097/GOX.0000000000002187. 
  6. Hong L, Shen M, Fang J, Wang Y, Bao Z, Bu S, Zhu Y. Hyaluronic acid (HA)-based hydrogels for full-thickness wound repairing and skin regeneration. J Mater Sci Mater Med. 2018 Sep 8;29(9):150. doi: 10.1007/s10856-018-6158-x.
  7. Mitran MI, Nicolae I, Tampa M, Mitran CI, Caruntu C, Sarbu MI, Ene CD, Matei C, Georgescu SR, Popa MI. Reactive Carbonyl Species as Potential Pro-Oxidant Factors Involved in Lichen Planus Pathogenesis. Metabolites. 2019 Oct 3;9(10):213. doi: 10.3390/metabo9100213. 
  8. Osikov MV, Gizinger OA, Ogneva OI. [Mechanism of the influence of melatonin on the immune status in experimental desynchronosis under led lighting]. Medical Immunology. 2015.17(6):517-524. [Article in Russian].
  9. Osikov MV, Telesheva LF, Ageev YuI. [Antioxidant effect of erythropoetin in experimental chronic renal failure]. Bull Exp Biol Med. 2015;160(8):162-165. [Article in Russian].
  10. Osikov MV, Makarov EV, Krivokhizhina LV. Hemostasiological effects of α1-acid glycoprotein in experimental septic peritonitis. Bull Exp Biol Med. 2007;144(8):143-145. [Article in Russian].
  11. Zhao D, Yu Y, Shen Y, Liu Q, Zhao Z, Sharma R, Reiter RJ. Melatonin Synthesis and Function: Evolutionary History in Animals and Plants. Front Endocrinol (Lausanne). 2019 Apr 17;10:249. doi: 10.3389/fendo.2019.00249.
  12. Markus RP, Fernandes PA, Kinker GS, da Silveira Cruz-Machado S, Marçola M. Immune-pineal axis - acute inflammatory responses coordinate melatonin synthesis by pinealocytes and phagocytes. Br J Pharmacol. 2018 Aug;175(16):3239-3250. doi: 10.1111/bph.14083.
  13. Varoni EM, Soru C, Pluchino R, Intra C, Iriti M. The Impact of Melatonin in Research. Molecules. 2016 Feb 20;21(2):240. doi: 10.3390/molecules21020240. 
  14. Slominski AT, Kim TK, Kleszczyński K, Semak I, Janjetovic Z, Sweatman T, Skobowiat C, Steketee JD, Lin Z, Postlethwaite A, Li W, Reiter RJ, Tobin DJ. Characterization of serotonin and N-acetylserotonin systems in the human epidermis and skin cells. J Pineal Res. 2020 Mar;68(2):e12626. doi: 10.1111/jpi.12626.
  15. Rusanova I, Martínez-Ruiz L, Florido J, Rodríguez-Santana C, Guerra-Librero A, Acuña-Castroviejo D, Escames G. Protective Effects of Melatonin on the Skin: Future Perspectives. Int J Mol Sci. 2019 Oct 8;20(19):4948. doi: 10.3390/ijms20194948.
  16. Shabeeb D, Najafi M, Musa AE, Keshavarz M, Shirazi A, Hassanzadeh G, Hadian MR, Samandari H. Biochemical and Histopathological Evaluation of the Radioprotective Effects of Melatonin Against Gamma Ray-Induced Skin Damage. Curr Radiopharm. 2019;12(1):72-81. doi: 10.2174/1874471012666181120163250.
  17. Li H, Yao Z, Tan J, Zhou J, Li Y, Wu J, Luo G. Epidemiology and outcome analysis of 6325 burn patients: a five-year retrospective study in a major burn center in Southwest China. Sci Rep. 2017 Apr 6;7:46066. doi: 10.1038/srep46066.
  18. Ageeva AA, Osikov MV, Simonjan EV, Toporec TA, Potehina EA, authors. Federal State Budgetary Educational Institution of Higher Education "South Ural State Medical University" of the Ministry of Health of the Russian Federation, patent holder. “Remedy in the form of a medicinal film containing melatonin for the treatment of thermal injury.”  Patent #2 751 048 07.07.2021. [In Russian].
  19. Volchegorsky IA, Nalimov AG, Yarovinsky VG. [Comparison of different approaches to the determination of LPO products in heptane-isopropanol blood extracts]. Questions of Medical Chemistry. 1989;(35)1:127-131.[Article in Russian].
  20. Davenport L, Dobson G, Letson H. A new model for standardising and treating thermal injury in the rat. MethodsX. 2019 Sep 12;6:2021-2027. doi: 10.1016/j.mex.2019.09.006. 
  21. Nagy B, Szélig L, Rendeki S, Loibl C, Rézmán B, Lantos J, Bogár L, Csontos C. Dynamic changes of matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 1 after burn injury. J Crit Care. 2015 Feb;30(1):162-6. doi: 10.1016/j.jcrc.2014.07.008. 
  22. Wilgus TA. Vascular Endothelial Growth Factor and Cutaneous Scarring. Adv Wound Care (New Rochelle). 2019 Dec 1;8(12):671-678. doi: 10.1089/wound.2018.0796. 
  23. de Aquino PEA, de Souza TFG, Santos FA, Viana AFSC, Louchard BO, Leal LKAM, Rocha TM, Evangelista JSAM, de Aquino NC, de Alencar NMN, Silveira EDR, Viana GSB. The Wound Healing Property of N-Methyl-(2S,4R)-trans-4-Hydroxy-L-Proline from Sideroxylon obtusifolium is Related to its Anti-Inflammatory and Antioxidant Actions. J Evid Based Integr Med. 2019 Jan-Dec;24:2515690X19865166. doi: 10.1177/2515690X19865166.
  24. Qin FJ, Hu XH, Chen Z, Chen X, Shen YM. Protective effects of tiopronin against oxidative stress in severely burned patients. Drug Des Devel Ther. 2019 Aug 13;13:2827-2832. doi: 10.2147/DDDT.S215927. 
  25. He F, Jiao H, Tian Y, Zhao L, Liao X, Fan Z, Liu B. Facile and large-scale synthesis of curcumin/PVA hydrogel: effectively kill bacteria and accelerate cutaneous wound healing in the rat. J Biomater Sci Polym Ed. 2018 Mar;29(4):325-343. doi: 10.1080/09205063.2017.1417002. 
  26. Olczyk P, Komosinska-Vassev K, Ramos P, Mencner Ł, Olczyk K, Pilawa B. Application of Numerical Analysis of the Shape of Electron Paramagnetic Resonance Spectra for Determination of the Number of Different Groups of Radicals in the Burn Wounds. Oxid Med Cell Longev. 2017;2017:4683102. doi: 10.1155/2017/4683102. 
  27. Nakazawa H, Ikeda K, Shinozaki S, Yasuhara S, Yu YM, Martyn JAJ, Tompkins RG, Yorozu T, Inoue S, Kaneki M. Coenzyme Q10 protects against burn-induced mitochondrial dysfunction and impaired insulin signaling in mouse skeletal muscle. FEBS Open Bio. 2019 Jan 19;9(2):348-363. doi: 10.1002/2211-5463.12580. 
  28. Lee YH, Bang ES, Lee JH, Lee JD, Kang DR, Hong J, Lee JM. Serum Concentrations of Trace Elements Zinc, Copper, Selenium, and Manganese in Critically Ill Patients. Biol Trace Elem Res. 2019 Apr;188(2):316-325. doi: 10.1007/s12011-018-1429-4. 
  29. Reiter RJ, Rosales-Corral S, Tan DX, Jou MJ, Galano A, Xu B. Melatonin as a mitochondria-targeted antioxidant: one of evolution's best ideas. Cell Mol Life Sci. 2017 Nov;74(21):3863-3881. doi: 10.1007/s00018-017-2609-7. 
  30. Osikov MV, Simonyan EV, Ageeva AA, Ageev YuI. [Melatonin in the dermal film limits the blood lymphocyte death in experimental thermal trauma]. Medical Immunology. 2021;23(2):389-394. [Article in Russian]
  31. Alluri H, Wilson RL, Anasooya Shaji C, Wiggins-Dohlvik K, Patel S, Liu Y, Peng X, Beeram MR, Davis ML, Huang JH, Tharakan B. Melatonin Preserves Blood-Brain Barrier Integrity and Permeability via Matrix Metalloproteinase-9 Inhibition. PLoS One. 2016 May 6;11(5):e0154427. doi: 10.1371/journal.pone.0154427. 
  32. Hazra S, Chaudhuri AG, Tiwary BK, Chakrabarti N. Matrix metallopeptidase 9 as a host protein target of chloroquine and melatonin for immunoregulation in COVID-19: A network-based meta-analysis. Life Sci. 2020 Sep 15;257:118096. doi: 10.1016/j.lfs.2020.118096.
  33. Qin W, Li J, Zhu R, Gao S, Fan J, Xia M, Zhao RC, Zhang J. Melatonin protects blood-brain barrier integrity and permeability by inhibiting matrix metalloproteinase-9 via the NOTCH3/NF-κB pathway. Aging (Albany NY). 2019 Dec 7;11(23):11391-11415. doi: 10.18632/aging.102537. 
  34. Rahbarghazi A, Siahkouhian M, Rahbarghazi R, Ahmadi M, Bolboli L, Keyhanmanesh R, Mahdipour M, Rajabi H. Role of melatonin in the angiogenesis potential; highlights on the cardiovascular disease. J Inflamm (Lond). 2021 Feb 2;18(1):4. doi: 10.1186/s12950-021-00269-5.
  35. Lee JH, Yoon YM, Han YS, Jung SK, Lee SH. Melatonin protects mesenchymal stem cells from autophagy-mediated death under ischaemic ER-stress conditions by increasing prion protein expression. Cell Prolif. 2019 Mar;52(2):e12545. doi: 10.1111/cpr.12545.

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Received October 18, 2021.
Accepted November 19, 2021.
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