The correlation of morphometric and biochemical parameters in regenerating tissues of skin burn wounds in rats at assession the reparative properties of 2-ethyl-6-methyl-3-hydroxypyridinium N-acetyl-6-aminohexanoate

Abstract

BACKGROUND: Aminohexanoic acid and its derivatives are potential reparants. The study is dedicated to the assessment of the aminohexanoic acid derivative on the healing of experimental thermal skin burns.

AIM: To study the relationship between morphometric and biochemical parameters during an experiment to evaluate the effectiveness of 2% ointment with 2-ethyl-6-methyl-3-hydroxypyridinium N-acetyl-6-aminohexanoate for the treatment of skin burns in rats.

MATERIALS AND METHODS: The thermal skin burns of 25 mm² area was modeled in 63 white non-linear sexually mature female rats under general anesthesia with Zoletil 100. The rats were divided into three groups depending on the local effect on the burn area: the control 1 group of rats — with imitation of application procedure without applying substances; the control 2 group with application of an ointment base (polyethylene glycol); the experimental group with application of 2% ointment with 2-ethyl-6-methyl-3-hydroxypyridinium N-acetyl-6-aminohexanoate. Applications of ointment, ointment base and their imitation were carried out starting from the second day until the end of the experiment. Morphometric and biochemical parameters were studied in homogenates of regenerating tissues of skin wounds on 7, 14 and 21 experiment days.

RESULTS: In experimental group rats during the inflammation phase, the efficiency of regeneration, confirmed by the dynamics of the area wounds, was largely correlated with the width of the leukocyte shaft and the thickness of granulation tissue, which, in turn, depended on the levels of Tumor Necrosis Factor alfa and Vascular Endothelial Growth Factor, respectively. In the proliferation phase, the area of wounds was significantly correlated with the length of the epithelial wedge and the fibroblasts number in the field of view, which, respectively, depended on the levels of Transforming Growth Factor beta and basic Fibroblast Growth Factor.

CONCLUSIONS: Results of the correlation analysis have testified that the reparative effect of local exposure with 2% ointment with 2-ethyl-6-methyl-3-hydroxypyridinium N-acetyl-6-aminohexanoate is ensured by activation of the synthesis of growth factors that stimulate the proliferation of capillaries and fibroblasts — components of granulation tissue, the active formation of which induces epidermis repair, which confirmed by planimetric data.

Keywords

thermal skin lesions / reparation / morphometry / cytokines / growth factors

References

[1] Kalinin RE, Suchkov IA, Mzhavanadze ND, et al. Regenerative technologies in the treatment of diabetic foot syndrome. Genes and cells. 2017;12(1):15–26. EDN: YUPZOV doi: 10.23868/201703002

[2] Калинин Р.Е., Сучков И.А., Мжаванадзе Н.Д. и др. Регенеративные технологии в лечении синдрома диабетической стопы // Гены и клетки. 2017. Т. 12, № 1. C. 15–26. EDN: YUPZOV doi: 10.23868/201703002

[3] Fayazov AD, Tulyaganov DB, Kamilov UR, Ruzimuratov DA. Contemporary local treatment of burn wounds. Bulletin of Emergency Medicine. 2019;12(1):43–47. EDN: DTFIUN

[4] Фаязов А.Д., Туляганов Д.Б., Камилов У.Р., Рузимуратов Д.А. Современные методы местного лечения ожоговых ран // Вестник экстренной медицины. 2019. Т. 12, № 1. С. 43–47. EDN: DTFIUN

[5] Imasheva AK, Lazko MV. Features of regenerative processes of the skin in thermal burns. Fundamental research. 2009;5:22–24. (In Russ.) EDN: KXPXUN

[6] Имашева А.К., Лазько М.В. Особенности регенераторных процессов кожи при термических ожогах // Фундаментальные исследования. 2009. № 5. C. 22–24. EDN: KXPXUN

[7] Kataev VA, Markov IA. The prospect of using new wound–healing medical products for the treatment of wounds and burns. In: Collection of scientific papers of the 4th Congress of combustiologists of Russia. 2013;49–50:102–104. (In Russ.) EDN: ZAGLPF

[8] Катаев В.А., Марков И.А. Перспектива использования новых ранозаживляющих медицинских средств для лечения ран и ожогов. В кн.: Сборник научных трудов 4-го съезда комбустиологов России. 2013. № 49–50. С. 102–104. EDN: ZAGLPF

[9] Naumkina VV. The experience of treating burn wounds with chitopran bandages. In: Proceedings of the All–Russian scientific and practical conference with international participation “Thermal injuries and their consequences”. 2016;56–57:137–138. (In Russ.)

[10] Наумкина В.В. Опыт лечения ожоговых ран повязками хитопран. В кн.: Материалы Всероссийской научно-практической конференции с международным участием «Термические поражения и их последствия». 2016. № 56–57. С. 137–138.

[11] Ostrovsky NV, Balyanina IB. Local conservative treatment of burn wounds using biomaterials developed by Saratov scientists. In: Modern aspects of thermal injury treatment. Proceedings of the scientific and practical conference with international participation dedicated to the 70th anniversary of the first burn center of Russia, Saint Petersburg, June 23-24, 2016. GBU Saint Petersburg Institute of Emergency Medicine named after I.I. Dzhanelidze; 2016. P. 74–75. (In Russ.) EDN: WIITQV

[12] Островский Н.В., Балянина И.Б. Местные консервативные лечения ожоговых ран с применением биоматериалов, разработанных саратовскими учеными. В кн.: Современные аспекты лечения термической травмы: материалы научно-практической конференции с международным участием, посвященной 70-летию первого ожогового центра России, Санкт-Петербург, 23–24 июня 2016 года. ГБУ Санкт-Петербургский НИИ скорой помощи им. И.И. Джанелидзе. 2016. С. 74–75. EDN: WIITQV

[13] Alekseeva NT. Morphological features of the wound process in the skin at the regional therapeutic influence [dissertation]. Orenburg; 2015. (In Russ.)

[14] Алексеева Н.Т. Морфологические особенности раневого процесса в коже при региональном лечебном воздействии: дис. ... д-ра мед. наук. Оренбург, 2015.

[15] Li Y, Xu DB, Wang HJ. Effects of hydrogen sulfide on the secretion of cytokines in macrophages of deep partial-thickness burn wound in rats. Zhonghua Shao Shang Za Zhi. 2016;32(7):408–412. doi: 10.3760/cma.j.issn.1009-2587.2016.07.005

[16] Li Y., Xu D.B., Wang H.J. Effects of hydrogen sulfide on the secretion of cytokines in macrophages of deep partial-thickness burn wound in rats // Zhonghua Shao Shang Za Zhi. 2016. Vol. 32, N 7. P. 408–412. doi: 10.3760/cma.j.issn.1009-2587.2016.07.005

[17] Dikici S, Yar M, Bullock AJ, et al. Developing wound dressings using 2-deoxy-D-ribose to induce angiogenesis as a backdoor route for stimulating the production of vascular endothelial growth factor. Int J Mol Sci. 2021;22(21):11437. doi: 10.3390/ijms222111437

[18] Dikici S., Yar M., Bullock A.J., et al. Developing wound dressings using 2-deoxy-D-ribose to induce angiogenesis as a backdoor route for stimulating the production of vascular endothelial growth factor // Int J Mol Sci. 2021. Vol. 22, N 21. P. 11437. doi: 10.3390/ijms222111437

[19] Elbialy ZI, Assar DH, Abdelnaby A, et al. Healing potential of Spirulina platensis for skin wounds by modulating bFGF, VEGF, TGF-ß1 and α-SMA genes expression targeting angiogenesis and scar tissue formation in the rat model. Biomed Pharmacother. 2021;137:111349. doi: 10.1016/j.biopha.2021.111349

[20] Elbialy Z.I., Assar D.H., Abdelnaby A., et al. Healing potential of Spirulina platensis for skin wounds by modulating bFGF, VEGF, TGF-ß1 and α-SMA genes expression targeting angiogenesis and scar tissue formation in the rat model // Biomed Pharmacother. 2021. Vol. 137, N 1. doi: 10.1016/j.biopha.2021.111349

[21] Makarevich PI, Efimenko AYu, Tkachuk VA. Biochemical regulation of regenerative processes by growth factors and cytokines: basic mechanisms and significance for regenerative medicine. Biochemistry (Moscow). 2020;85(1):11–26. EDN: VJCHNF doi: 10.1134/S0006297920010022

[22] Макаревич П.И., Ефименко А.Ю., Ткачук В.А. Биохимическая регуляция регенеративных процессов факторами роста и цитокинами: основные механизмы и значимость для регенеративной медицины // Биохимия. 2020. Т. 85, № 1. С. 15–33. EDN: OPVJOE doi: 10.31857/S0320972520010029

[23] Gan D, Su Q, Su H, et al. Burn ointment promotes cutaneous wound healing by modulating the PI3K/AKT/mTOR signaling pathway. Front Pharmacol. 2021;12:631102. doi: 10.3389/fphar.2021.631102

[24] Gan D., Su Q., Su H., et al. Burn ointment promotes cutaneous wound healing by modulating the PI3K/AKT/mTOR signaling pathway // Front Pharmacol. 2021. Vol. 12. P. 631102. doi: 10.3389/fphar.2021.631102

[25] Pakhomov DV, Blinova EV, Shimanovsky DN, et al. Evidence-based aspects of stimulating uncomplicated wounds healing with local use of acexamic acid silver salt. Operative surgery and clinical anatomy. 2020;4(1):19–25. EDN: SPMTIJ doi: 10.17116/operhirurg2020401119

[26] Пахомов Д.В., Блинова Е.В., Шимановский Д.Н. и др. Доказательные аспекты стимулирования заживления неосложненной раны при локальном применении серебряной соли ацексамовой кислоты // Оперативная хирургия и клиническая анатомия. 2020. Т. 4, № 1. С. 19–25. EDN: SPMTIJ doi: 10.17116/operhirurg2020401119

[27] Andrianova EV. Biochemical aspects of the pro-regenerative effect of a new derivative of N-acetyl-6-aminohexanoic acid [dissertation]. Tver; 2023. 139 p. (In Russ.) EDN: JQFLEA

[28] Андрианова Е.В. Биохимические аспекты прорегенераторного действия нового производного N-ацетил-6-аминогексановой кислоты: дис. ... канд. биол. наук. Тверь, 2023. 139 с. EDN: JQFLEA

[29] Peretyagin SP, Martusevich AK, Grishina AA, et al. Laboratory animals in experimental medicine. Nizhnii Novgorod: Privolzhsky Research Medical University; 2011. 300 p. (In Russ.) EDN: SIXODX

[30] Перетягин С.П., Мартусевич А.К., Гришина А.А. и др. Лабораторные животные в экспериментальной медицине. Нижний Новгород: Нижегородский научно-исследовательский институт травматологии и ортопедии, 2011. 300 c. EDN: SIXODX

[31] Ogneva NS, Savchenko ES, Taboyakova LA. Anesthesia of female mice during surgical embryo transfer. Journal Biomed. 2021;17(3E):64–69. EDN: GQUYTV doi: 10.33647/2713-0428-17-3E-64-69

[32] Огнева Е.С., Савченко Е.С., Табоякова Л.А. Анестезия самок мышей при хирургической трансплантации эмбрионов // Биомедицина. 2021. Т. 17, № S3. С. 64–69. EDN: GQUYTV doi: 10.33647/2713-0428-17-3E-64-69

[33] Fine AM, Petukhova MN, Miguleva IYu, Savotchenko AM. Comparative evaluation of two schemes of intramuscular anesthesia in laboratory rats in an experiment. Issues of Reconstructive and Plastic Surgery. 2019;22(2):53–61. EDN: OTJJTX doi: 10.17223/1814147/69/07

[34] Файн А.М., Петухова М.Н., Мигулева И.Ю., Савотченко А.М. Сравнительная оценка двух схем внутримышечного наркоза у лабораторных крыс в эксперименте // Вопросы реконструктивной и пластической хирургии. 2019. Т. 22, № 2. С. 53–61. EDN: OTJJTX doi: 10.17223/1814147/69/07

[35] Petrovskaya MA. Preclinical evaluation of the effectiveness of the reparative properties of 2-ethyl-6-methyl-3-hydroxypyridinium N-acetyl-6-aminohexanoate. Tver Medical Journal. 2023;6:49–52. (In Russ.) EDN: MQWKYI

[36] Петровская М.А. Доклиническая оценка эффективности репаративных свойств 2-этил-6-метил-3-гидроксипиридиния N-ацетил-6-аминогексаноата // Тверской медицинский журнал. 2023. № 6. С. 49–52. EDN: MQWKYI

[37] Pakhomova AE, Pakhomova YuV, Pakhomova EE. A new method of experimental modeling of thermal skin burns in laboratory animals that meets the principles of Good Laboratory Practice. Journal of Siberian Medical Sciences. 2015;3:97–100. EDN: VXOLBT

[38] Пахомова А.Е., Пахомова Ю.В., Пахомова Е.Е. Новый способ экспериментального моделирования термических ожогов кожи у лабораторных животных, отвечающий принципам Good Laboratory Practice (надлежащей лабораторной практики) // Медицина и образование в Сибири. 2015. № 3. C. 97–100. EDN: VXOLBT

[39] Semchenko VV, Barashkova SA, Nozdrin VI, Artemiev VN. Histological technique. Textbook. 3d ed. Omsk; Oryol: Omskaya oblastnaya tipografiya; 2006. (In Russ.)

[40] Семченко В.В., Барашкова С.А., Ноздрин В.И., Артемьев В.Н. Гистологическая техника: учебное пособие для студентов медицинских вузов. 3-е изд. Омск; Орел: Омская областная типография, 2006.

[41] Petrovskaya MA, Petrova MB, Egorova EN, Andrianova EV. Morphology of regeneration phases and dynamics of growth factor levels in the application of 2-ethyl-6-methyl-3-hydroxypyridinium N-acetyl-6-aminohexanoate for the repair of thermal burns of rat skin. Medical Academic Journal. 2023;23(3):21–29. EDN: MJGYMP doi: 10.17816/MAJ606647

[42] Петровская М.А., Петрова М.Б., Андрианова Е.В., Егорова Е.Н. Особенности морфологии фаз регенерации и динамика уровней факторов роста при применении 2-этил-6-метил-3-гидроксипиридиния N-ацетил-6-аминогексаноата для репарации термических ожогов кожи крыс // Медицинский академический журнал. 2023. Т. 23, № 3. С. 21–29. EDN: MJGYMP doi: 10.17816/MAJ606647

[43] Petrovskaya MA, Petrova MB, Andrianova EV, Egorova EN. Dynamics of cytokine levels in regenerating tissues of thermal burns of rat skin when applying ointment with a new derivative of N-acetylaminohexanoic acid. Genes and Cells. 2022;17(3):178–179. (In Russ.) EDN: CJOMZS

[44] Петровская М.А., Петрова М.Б., Андрианова Е.В., Егорова Е.Н. Динамика уровней цитокинов в регенерирующих тканях термических ожогов кожи крыс при применении мази с новым производным N-ацетиламиногексановой кислоты // Гены и клетки. 2022. Т. 17, № 3. С. 178–179. EDN: CJOMZS

[45] Omel’yanenko NP, Slutskii LI. Connective tissue (histophysiology and biochemistry). Mironov SP, ed. Moscow: Izvestiya; 2009. (In Russ.) EDN: QKSAIH

[46] Омельяненко Н.П., Слуцкий Л.И. Соединительная ткань (гистофизиология и биохимия). Москва: Известия, 2009. EDN: QKSAIH

[47] El Ayadi A, Jay JW, Prasai A. Current approaches targeting the wound healing phases to attenuate fibrosis and scarring. Int J Mol Sci. 2020;21(3):1105. doi: 10.3390/ijms21031105

[48] El Ayadi A., Jay J.W., Prasai A. Current approaches targeting the wound healing phases to attenuate fibrosis and scarring // Int J Mol Sci. 2020. Vol. 21, N 3. P. 1105. doi: 10.3390/ijms21031105

[49] Ahn HN, Kang HS, Park SJ, et al. Safety and efficacy of basic fibroblast growth factors for deep second-degree burn patients. Burns. 2020;46(8):1857–1866. doi: 10.1016/j.burns.2020.06.019

[50] Ahn H.N., Kang H.S., Park S.J., et al. Safety and efficacy of basic fibroblast growth factors for deep second-degree burn patients // Burns. 2020. Vol. 46, N 8. P. 1857–1866. doi: 10.1016/j.burns.2020.06.019

[51] Otani Y, Komura M, Komura H, et al. Optimal amount of basic fibroblast growth factor in gelatin sponges incorporating β-tricalcium phosphate with chondrocytes. Tissue Eng Part A. 2015;21(3–4):627–636. doi: 10.1089/ten.TEA.2013.0655

[52] Otani Y., Komura M., Komura H., et al. Optimal amount of basic fibroblast growth factor in gelatin sponges incorporating β-tricalcium phosphate with chondrocytes // Tissue Eng Part A. 2015. Vol. 21, N 3–4. P. 627–636. doi: 10.1089/ten.TEA.2013.0655

[53] Chakrabarti S, Mazumder B, Rajkonwar J, et al. bFGF and collagen matrix hydrogel attenuates burn wound inflammation through activation of ERK and TRK pathway. Sci Rep. 2021;11(1):3357. doi: 10.1038/s41598-021-82888-9

[54] Chakrabarti S., Mazumder B., Rajkonwar J., et al. bFGF and collagen matrix hydrogel attenuates burn wound inflammation through activation of ERK and TRK pathway // Sci Rep. 2021. Vol. 11, N 1. P. 3357. doi: 10.1038/s41598-021-82888-9

[55] Tran-Nguyen TM, Le KT, Nguyen LT, et al. Third-degree burn mouse treatment using recombinant human fibroblast growth factor 2. Growth Factors. 2020;38(5–6):282–290. doi: 10.1080/08977194.2021.1967342

[56] Tran-Nguyen T.M., Le K.T., Nguyen L.T., et al. Third-degree burn mouse treatment using recombinant human fibroblast growth factor 2 // Growth Factors. 2020. Vol. 38, N 5–6. P. 282–290. doi: 10.1080/08977194.2021.1967342

[57] Nagaraja S, Chen L, DiPietro LA, et al. Predictive approach identifies molecular targets and interventions to restore angiogenesis in wounds with delayed healing. Front Physiol. 2019;10:636. doi: 10.3389/fphys.2019.00636

[58] Nagaraja S., Chen L., DiPietro L.A., et al. Predictive approach identifies molecular targets and interventions to restore angiogenesis in wounds with delayed healing // Front Physiol. 2019. Vol. 10. P. 636. doi: 10.3389/fphys.2019.00636

[59] Rahman MS, Islam R, Rana MM, et al. Characterization of burn wound healing gel prepared from human amniotic membrane and aloe vera extract. BMC Complement Altern Med. 2019;19:115. doi: 10.1186/s12906-019-2525-5

[60] Rahman M.S., Islam R., Rana M.M., et al. Characterization of burn wound healing gel prepared from human amniotic membrane and aloe vera extract // BMC Complement Altern Med. 2019. Vol. 19. P. 115. doi: 10.1186/s12906-019-2525-5

[61] Obraztsova AE, Nozdrevatykh AA. Morphofunctional features of the reparative process during healing of skin wounds, taking into account possible scar deformations (literature review). Journal of new medical technologies, eEdition. 2021;1:98–107. EDN: DNUEYX doi: 10.24412/2075-4094-2021-1-3-3

[62] Образцова А.Е., Ноздреватых А.А. Морфофункциональные особенности репаративного процесса при заживлении кожных ран с учетом возможных рубцовых деформаций (обзор литературы) // Вестник новых медицинских технологий. Электронное издание. 2021. № 1. C. 98–107. EDN: DNUEYX doi: 10.24412/2075-4094-2021-1-3-3

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