Objective : To substantiate a possibility for modification of inflammation process in thermal burns of stage II using nanoparticles of silver and proteinase inhibitors. Methods: The experiment included 30 mature Wistar male rats. After a thermal burn simualtion, 10 animals were locally applied with a water-soluble composition of silver nanoparticles in a sodium alginate matrix, 10 – with a combination of a water-soluble composition of nanosilver and aprotinin (another 10 rats were control). After 7 days, animals were withdrawn from the experiment and were run histological and immunohistochemical examination of the skin in the burn area. Results : The maximum effect of combined wound therapy with a water-soluble composition of silver nanoparticles in a matrix of sodium alginate with a proteinase inhibitor (aprotinin), which contributed to the activation of regenerative processes, was revealed. Conclusions : The effect of combined therapy is related to an improvement in the balance of proliferation and apoptosis in a burn wound as well as with a specific stimulation of the inflammatory response. This local therapy for burns can be considered pathogenically justified and promising.

The goal was to prove by complex morphological analysis the purposefulness of using a biocomposition based on silver nanoparticles stabilized in solution with sodium alginate and the inhibitor of proteases of aprotinin for influencing the course of the inflammatory process after thermal burn.Using light and electron microscopy we studied morphological changes after thermal skin damage of II grade in rats using biocomposite based on silver nanoparticles stabilized in solution with sodium alginate and the protease inhibitor aprotinin to modify the course of the inflammatory process. Research was performed on 80 white male Wistar rats, 180-200g weight.The early wound process (3rd day) did not differ in various groups and was characterized by the predominance of exudative-necrotic and dyscirculatory disorders over the regeneration processes. From the 7th day, the morphological picture began to differ depending on the applied medication therapy. There was noted the beginning of rejection of the scab, a decrease in the intensity of inflammation, in hemodynamic disturbances and the onset of epithelialization. On the 14th day, the epithelialization eras observed, the inflammatory process was decreasing and the young scar was forming. The use of erythromycin facilitated the migration of keratinocytes and accelerated the restoring of connective tissue in dermis through synthetic activity of fibroblasts. The use of the silver nanoparticle composition had a moderate corrective effect. After proteinase inhibitors application, the dystrophic changes, both in the epidermis and in dermis, persisted longer than in other groups. At the same time, samples where burns were treated by combined way (the solution of nano-silver and an inhibitor of proteases of aprotinin) demonstrated morphological signs of the increase in the keratinocytes proliferative activity and functional activation of protein synthesis of fibroblasts. According to the results of light and electron microscopy, the best result was obtained after combined therapy; the morphological characteristic of wounds was somewhat worse after antibiotic or silver monotherapy. The monotherapy only with the proteinase inhibitor gave the worst results on the duration of healing and the severity of inflammation in the wound.The local application of the silver nanoparticles stabilized in sodium alginate solution and aprotinin limited the destructive changes in the burnt skin, assisted in early repair processes, helped in the formation of granulation tissu e. its maturation and epithelization.

Актуальность. Ожоговые повреждения кожи, в частности термические ожоги, во всем мире являются общепризнанной социально-медицинской проблемой. Согласно данным ВОЗ, на долю термических ожогов кожи и ассоциированных с ними осложнений приходится около 15% среди общего числа травматических повреждений мирного времени. Несмотря на быстрое внедрение новых методов лечения ожогов кожи, вопрос эффективности этиопатогенетически обоснованной терапии остается актуальным и дискутабельным. Цель. Определить степень и характер изменений неспецифических протеиназ и их ингибиторов в сыворотке крови и супернатантах гомогенатов кожи у крыс линии Wistar с моделью термического ожога кожи II степени, а также на фоне применения биокомпозиции на основе наночастиц серебра, стабилизированных в растворе альгинатом натрия, и ингибитора протеиназ апротинина для оценки модификации течения воспалительного процесса. Методы. Проведено две серии экспериментальных исследований на белых крысах самцах линии Wistar: в первой серии оценивали динамику изменений в протеиназ-ингибиторной системе сыворотки крови и супернатантах гомогенатов кожи крыс без коррекции, а во второй серии изучали вышеперечисленные изменения на фоне медикаментозной коррекции. В сыворотке и супернатантах гомогенатов энзиматическими методами определяли трипсиноподобную активность (ТПА), эластазоподобную активность (ЭПА), антитриптическую активность (АТА) и кислотостабильные ингибиторы (КСИ); белок определяли методом Лоури. Результаты. В статье представлены результаты динамических изменений показателей неспецифических протеиназ и их ингибиторов при моделировании термического ожога II степени на 100 белых крысах линии Wistar. Наиболее эффективным вариантом экспериментальной коррекции нарушений протеиназ-ингибиторного гомеостаза является сочетанное применение аппликаций с раствором наносеребра и апротинина. Заключение. Полученные данные изменений протеолитической активности отражают характер выраженности воспалительного процесса и могут свидетельствовать о развитии деструктивных изменений в кожных покровах крыс. Установлено, что на локальном уровне дисбаланс неспецифических протеиназ и их ингибиторов характеризуется активацией протеиназ на фоне истощения ингибиторного потенциала, в то время как на системном - повышение протеолитической активности сопровождается ростом уровней ингибиторов, что подтверждает целесообразность локальной терапии ожогов с применением ингибиторов протеиназ. Наиболее эффективным вариантом коррекции нарушений протеиназ-ингибиторного гомеостаза является сочетанное применение аппликаций с раствором наносеребра и апротинина. Background. Burn injuries of the skin, specifically thermal burns, are internationally recognized as an important social and medical issue. According to statistical data of the World Health Organization thermal skin burns and their complications account for approximately 15% of all peacetime traumatic injuries. Despite the rapid development of new therapies for skin burns the effectiveness of etiologically and pathogenetically substantiated therapy remains a relevant and controversial issue. Aim: To determine the degree and nature of changes in nonspecific proteinases and their inhibitors in serum and supernatants of skin homogenates from Wistar rats with second degree deep thermal burn wounds and during the use of a biocomposition based on silver nanoparticles stabilized in a solution with sodium alginate and a proteinase aprotinin inhibitor for modification of the inflammatory process. Methods. Two series of experiments were performed on Wistar male rats: 1) evaluating the dynamics of changes in proteinases and their inhibitors in blood serum and supernatants of rat skin homogenates without correction and 2) studying the above-listed changes during a pharmaceutical correction. The trypsin-like activity (TLA), elastase-like activity (ELA), antitryptic activity (ATA), and acid-stable inhibitors (ASI) were measured in serum and supernatants of homogenates using enzymatic methods; protein concentration was determined by the Lowry method. Results. The article presents dynamic changes in nonspecific proteinases and their inhibitors studied on a model of second degree deep thermal burn wounds in 100 Wistar rats. Combination applications with a solution of nanosilver and aprotinin was the most effective experimental treatment of the proteinase and inhibitor homeostasis disorders. Conclusions. The observed changes in proteolytic activity reflect the nature of inflammatory process and may indicate development of destructive changes in the rat skin and alleviation of inflammation using the experimental treatment. At the local level, the imbalance of nonspecific proteinases and their inhibitors was characterized by activation of proteases associated with depletion of the inhibitory potential while at the systemic level, the increase in proteolytic activity was associated with a reactive increase in levels of inhibitors, which confirms the advisability of local therapy for burns with proteinase inhibitors. The most effective experimental therapy for the proteinase and inhibitor homeostasis disorders was the combined use of applications with a solution of nanosilver and aprotinin.

Aim. The study was designed to perform a complex examination of the dynamics of the activity of protease inhibitor system along with the morphological study of cell and tissue components of the skin regeneration in the modelling of the 2nd degree thermal burn in rats.Materials and methods. The studies were conducted on 40 mature male Wistar rats. We determined the trypsin-like, elastase-like, antitrypsin activity and acid-stable inhibitors of proteinases in supernatants of homogenates of the skin of rats after the modelling of a burn injury by the enzymatic methods. The wound healing process was evaluated based on the macroscopic, histological examination with the general staining by hematoxylin and eosin, van Gieson’s picrofuchsin, and transmission electron microscopy.Results. The complex analysis of the skin regeneration revealed the role of the enzymatic reactions and the inflammatory microenvironment in the development of pathogenetic and morphological tissue changes in the dynamics of wound healing. The homeostasis of proteinase-inhibitory system was accompanied by both local and systemic disorders. The nonspecific proteinases occupied the key positions among the mediators of the biochemical alteration. The dynamics of changes was characterized by a powerful proteolysis with a progressive depletion of the inhibitory potential, which caused the development of a secondary-alterative process. The launch of the cascade of the inflammatory responses led to the disruption of the microcirculation and the damage of the blood-tissue barriers, the proteolytic destruction of the connective fibers, the formation of an edema and the disruption of the regenerative capacity of a damaged tissue. Morphological destructive changes in the skin remained up to 7-14 days, a decreased regenerative ability of the skin was manifested by a violation of the proliferation and differentiation of the epidermis. According to the electron microscopy, the phenomenon of tissue and intracellular edema, lesions of the microcirculatory bed, leukocyte-macrophage infiltration persisted up to 14 days of the experiment. The disruptions of the structure of the intercellular contacts between epidermocytes, macrophages in the dermis and many newly formed collagen fibers were still noticeable by the 14th day. Conclusion. The results of the study showed the progressive development of the destructive reactions after the modelling of 2nd degree thermal burn. There was an increase in the level of proteolytic activity on the background of depletion of the inhibitory potential. There was the persistence of the alterative changes in the skin up to 7-14 days and the complication of the reparative processes with suppuration, impaired proliferation and differentiation of the epidermis. The performed investigation is important for the determination of further ways of pathogenetic correction.