scholarly journals Electron microscopic changes of the testis germinal epithelium after an experimental thermal trauma in the application of cryo-lyophilized xenograft skin substrate

2018 ◽  
Vol 24 (4) ◽  
pp. 53-57
Author(s):  
K.S. Volkov ◽  
S.Yu. Muha

Deep, large area of thermal trauma of the skin leads to significant morphofunctional changes in organs in the composition of the burned organism. However, condition of the central organ of the male reproductive system - testicle after burns and, especially, in the treatment of thermally damaged areas of the skin remain poorly understood. The purpose of this study was to establish an ultrastructural re-organization of testis germinal epithelium cells in the stages of toxemia and septicotoxemia following an experimental thermal trauma when using xenograft skin substrate. The studies were carried out on 35 sexually mature white male rats, which were divided into three groups: 1 - intact animals (5 rats), 2 - animals with burn injury (15 rats), 3 - animals with burn injury, for which were used xenograft skin substrate (15 rats). The burn was applied under ketamine anesthesia with copper plates, heated in boiling water, on 1820% shaved body surface of animals. Early necrectomy of the damaged areas of skin was carried out 1 day after the thermal burn. The formed wounds were covered with cryo-lyophilized xenograft skin substrate. Experimental animals were decapitated at 7, 14 and 21 days of the experiment (early, late toxemia and septicotoxemia). The material sampling for electron microscopy was carried out in accordance with a generally accepted methodology. It is established that in the stage of early toxemia (7 days of experiment) there are adaptive-compensatory changes and signs of destruction of spermatogenic cells of testis. In the stages of late toxemia and septicotoxemia (14 and 21 days of the trial), significant destructive changes occur in all components of the seminiferous tubules of the organ. Electron microscopically found that the closure of the burn wound after the early necrectomy of cryo-lyophilized xenograft skin substrate in severe experimental burn injury in the early stages of the experiment significantly reduces the damage of testis germinal epithelium of experimental animals and activates regenerative processes. This contributes to a significant improvement in the ultrastructure of the components of the convoluted tubules of the organ in the late stages of the experiment.


2019 ◽  
Vol 18 (4) ◽  
pp. 17-23
Author(s):  
S. Yu. Mukha

The aim of this work was to establish the histological and morphometric changes of the structural components of the rat testes after experimental thermal trauma and under condition of correction. Object and methods. The study was performed on 48 sexually mature outbred white male rats following the rules of bioethics. The experimental animals were divided into three groups: intact; with severe thermal trauma; with burn injury and application of xenodermal substrate after early necrectomy of the damaged tissues. Results and discussion. Already on the 7th day after application of xenodermal substrate the processes of reparative regeneration of hemocapillaries is activated, the degree of vascular disorders and damages of the structural components of the testes are decreased, and regenerative processes are activated. The percentage of significantly altered seminiferous tubules is 1.90-fold lower than in animals without correction. In the later stages of the experiment, the usage of the xenodermal substrate contributes to an active flow of regenerative processes and a relative normalization of all the structural components of the testes. Morphometric indicators of mean values of diameter and area of convoluted tubes on the 21th day were significantly (p<0.001) higher by 1.39 times and 2.00 times relative to the indicators in the group of animals without correction. Areas with unchanged histostructure are dominated in the testes at this term. The percentage of significantly altered tubules was 10.58 times lower than the corresponding value in animals with burn injury without correction. Conclusions. Thus, the application of a xenodermal substrate on a wound formed after necrectomy of thermally damaged areas was found to contribute to a significant reduction of vascular disorders and destructive changes of spermatogenic cells. Better state of intracellular components and activation of regeneration contribute to a relative normalization of the testicular structure and morphometric indices in the later terms of the experiment.



Author(s):  
O. I. Tiron

Despite the fact that the main pathogenetic factor in the development of burn disease is the alteration of the skin, the severity of the condition of patients in the acute period of the disease is due to systemic changes in the body caused by thermal factor. Being a powerful stress factor, severe burns are accompanied by activation of the hypothalamic-pituitary system. Although thyroid hormones are not considered typical "stress hormones", they also play a role in maintaining vasomotor function under stress. The question of thyroid dysfunction on the background of burn injury remains debatable, and scientific data on its morphological and ultramicroscopic changes are quite limited. The aim of the work is to establish microscopic and submicroscopic changes of the thyroid gland of experimental animals in the stage of shock after thermal trauma of the skin. Experimental studies were performed on 12 white adult male rats, which received a thermal burn of the skin of 2-3 degrees with a lesion area of 21-23 % of the skin. An infusion of 0.9 % NaCl solution at a dose of 10 ml/kg was performed into the inferior vena cava. Collection of material for histological and electron microscopic examinations was performed according to generally accepted methods. Histological specimens were examined using a MIKROmed SEO SCAN light microscope and photo-documented using a Vision CCD Camera with an image output system from histological specimens. Ultrathin sections made on an ultramicrotome UMPT-7 were contrasted with uranyl acetate, lead citrate according to the Reynolds method and studied under an electron microscope PEM-125K. Conducted micro- and submicroscopic studies of the structural components of the thyroid gland a day after the reproduction of thermal trauma to the skin on the background of the introduction of 0.9 % NaCl solution revealed reactive adjunctive-compensatory changes and initial manifestations of destruction. Vascular, stromal and parenchymal components of the organ in the stage of shock after burns correspond to the state of "stress" as a reaction to pathological exogenous exposure, and the intensification of metabolic processes in the affected organism of experimental animals.



2019 ◽  
Vol 25 (3) ◽  
pp. 16-20
Author(s):  
Z.M. Nebesna ◽  
O.I. Bashynska ◽  
N.P. Ocheretna ◽  
G.М. Galunko ◽  
O.Ya. Slyvka

Deep, large thermal burns are not limited to local lesions of tissues, they cause significant disruption of all systems and organs of the organism, change in metabolic processes. It is revealed that the primary links in the pathogenesis of burn disease are destruction of the skin, impaired neuroendocrine regulation and significant hemodynamic disorders. The reorganization of structures and impaired lung function, in response to a pathological process in the body, is attracting increasing attention of scientists. The aim of the study was to establish a submicroscopic rearrangement of the alveoli after a thermal lesion for 1 day after the experimental thermal trauma. Grade III burns were applied under ketamine anesthesia with copper plates heated in boiled water to a temperature of 97-1000С. The size of the lesion area was 18-20 % of the epilated surface of the body of rats. An experimental study of the structural components of lung alveoli after burn injury was performed on laboratory white male rats weighing 160-180 g. Euthanasia of rats was performed after ketamine anesthesia by decapitation. In the experiment, the study of the submicroscopic state of the walls of the alveoli of the lungs after thermal trauma was done. It is established that in the stage of shock after the application of burn injury – 1 day, in the alveoli of the respiratory department of the lung, there are adaptive compensatory and initial destructive changes of all structural components of the alveoli. Damage to the ultrastructure of the aerohematical barrier is manifested by intracellular edema and edema of the organelles of the endothelial cells, respiratory and secretory epitheliocytes, and the amount of heterochromatin increases in their deformed nuclei. The basement membrane also has signs of edema, sometimes homogeneous, fuzzy. The decrease in the number of vesicles and micropinocytotic vesicles in endothelial and respiratory epitheliocytes leads to impaired endothelial and alveolar metabolism. Numerous actively phagocytic alveolar macrophages with a well-expressed lysosomal apparatus are found in the alveoli. Initial alternative alterations of the ultrastructure of the components of the air-barrier barrier lead to disruption of gas exchange in the respiratory department of the lungs



2018 ◽  
Vol 22 (4) ◽  
pp. 610-615
Author(s):  
V.G. Korytskyi

Thermal trauma and burn disease, which develops from deep, large lesions, are accompanied by significant morphological and functional changes not only in the skin, but also in organs of all systems of the body, including thyroid gland. The purpose of the study was to establish a microscopic reorganization of the animal thyroid gland in dynamics after an experimental thermal trauma. Burns of the third degree were applied under ketamine anesthesia with copper plates heated in boiled water to a temperature of 97–100 °С. The affected area corresponded 18–20% of the surface of the rat’s body. Experimental study of structural changes in the vascular bed of the thyroid gland after severe thermal trauma was carried out on laboratory white male rats weighing 160–180 g. Euthanasia of rats was performed after ketamine anesthesia, by decapitation. Structural changes in the thyroid gland were studied after 1, 7, 14 and 21 days from the beginning of the experiment. Histologic sections of 5–6 µm thickness were stained with hematoxylin-eosin. For the study of the connective tissue condition and its main substance, silver impregnation was performed by using the Gordon-Sweet's method (to detect reticular and immature collagen fibers), staining with 3-colored MSB — selective coloring method for intravascular coagulation syndrome. It has been established that in the early stages after the thermal trauma (1–7 days of the experiment, the stage of shock and early toxemia) there are adaptive-compensatory changes and initial signs of destruction of the vascular bed structural components of the organ. In late terms (14–21 days of the experiment, the stages of late toxemia and septicotoxemia) significant destructive-degenerative changes of the thyroid gland arteries, veins and blood vessels of the hemocirculatory bed were detected. Therefore, a severe burn injury causes deep histological modification of the thyroid gland vessels, the degree of intensity of which depends on the time interval of the experiment, which adversely affects on the functioning of the organ and transvascular metabolic processes. In further researches it is planned to study the course of morphological changes of thyroid gland vessels during thermal trauma with combined application of corrective methods.  



Author(s):  
Vladislav A Dolgachev ◽  
Susan Ciotti ◽  
Emma Liechty ◽  
Benjamin Levi ◽  
Stewart C Wang ◽  
...  

Abstract Objective Burn wound progression is an inflammation driven process where an initial partial-thickness thermal burn wound can evolve over time to a full-thickness injury. We have developed an oil-in-water nanoemulsion formulation (NB-201) containing benzalkonium chloride for use in burn wounds that is antimicrobial and potentially inhibits burn wound progression. We used a porcine burn injury model to evaluate the effect of topical nanoemulsion treatment on burn wound conversion and healing. Methods Anesthetized swine received thermal burn wounds using a 25cm 2 surface area copper bar heated to 80 oC. Three different concentrations of NB-201 (10%, 20%, or 40% nanoemulsion), silver sulfadiazine cream or saline were applied to burned skin immediately after injury and on days 1, 2, 4, 7, 10, 14, and 18 post-injury. Digital images and skin biopsies were taken at each dressing change. Skin biopsy samples were stained for histological evaluation and graded. Skin tissue samples were also assayed for mediators of inflammation. Results Dermal treatment with NB-201 diminished thermal burn wound conversion to a full-thickness injury as determined by both histological and visual evaluation. Comparison of epithelial restoration on day 21 showed that 77.8% of the nanoemulsion treated wounds had an epidermal injury score of 0 compared to 16.7% of the silver sulfadiazine treated burns (p=0.01). Silver sulfadiazine cream and saline treated wounds (controls) converted to full-thickness burns by day 4. Histological evaluation revealed reduced inflammation and evidence of skin injury in NB-201 treated sites compared to control wounds. The nanoemulsion treated wounds often healed with complete regrowth of epithelium and no loss of hair follicles (NB-201: 4.8±2.1, saline: 0±0, silver sulfadiazine: 0±0 hair follicles per 4mm biopsy section, p&lt;0.05). Production of inflammatory mediators and sequestration of neutrophils were also inhibited by NB-201. Conclusions Topically applied NB-201 prevented the progression of a partial-thickness burn wound to full-thickness injury and was associated with a concurrent decrease in dermal inflammation.



2021 ◽  
pp. 113-116
Author(s):  
E.E. Savelieva ◽  
◽  
O.F. Veselova ◽  
I.V. Gackih ◽  
V.A. Horgevski ◽  
...  

The aim of the research. To investigate the anti-infl ammatory eff ect of Cinquefoil on a model of a thermal wound. Material and methods. A liquid extract in 70 % ethyl alcohol was obtained from dry crushed goose cinquefoil by the percolation method. Th e resulting extracts were evaporated under vacuum at a temperature of 50 °C. Th e dry residue was dissolved in water immediately before treatment at a dose of 250 mg / kg. Th e evaluation of anti-infl ammatory activity was carried out in an experiment on 20 white male mice. Th ermal skin burns were simulated in all animals. Th e burns were healed in an open way. During the entire experiment, the general condition of the experimental animals was assessed, body weight and temperature were determined, the appearance of wounds, appetite, and activity were assessed. For histological examination, a section of the skin was taken, including the burn zones and intact tissues. Paraffi n sections were stained with hematoxylin and eosin. Results. An analysis of the results of the experimental study showed that all experimental animals, under the infl uence of high temperatures, developed a third degree thermal burn of the skin, characterized by damage to the entire thickness of the skin with complete death of hair follicles, sweat and sebaceous glands. At the site of the burn, the skin was dense, motionless and insensitive to painful stimuli. Th e appearance of the wounds did not diff er between groups of animals. Comparative microscopy of the control and experimental groups showed that the depth of damage to the epidermis and dermis was more pronounced in the experimental group. In addition, in the same group, lymphocytic infi ltration of the dermis, hypodermis, the presence of a pronounced vascular reaction and edema of the dermis are more pronounced, which indicates a more pronounced infl ammatory. Conclusion. Th e analysis of the results obtained shows that in the treatment of burn wounds, the local application of the preparation Potentilla goose on the 4th day in the experiment does not lead to a pronounced anti-infl ammatory eff ect. However, the revealed lymphocytic infi ltration of the dermis and other histological parameters indicate more active processes in the tissues, which may subsequently lead to more accelerated wound cleansing and faster regeneration



2019 ◽  
Vol 9 (3-4) ◽  
pp. 163-171
Author(s):  
Heba A. Yassa ◽  
Randa T. Hanna ◽  
Hala Zein El Abdin

Background: Determination of time passed since burn injury in the living is critical in forensic science. Autophagy biomarkers and vitronectin can play an important role in determination of the age of burn injuries through their levels in the tissue. Objective: The aim of this study was to investigate the role of autophagy biomarkers in dating burn injury and to correlate them with the histopathological effects of deep second-degree thermal burn. Method: Fifty-four male rats were used in this study after infliction of second-degree thermal burns to their skin. Samples were taken from them after 30 minutes and one, four, 24, 48, and 72 hours following burn to be examined histologically and also for autophagy biomarkers and vitronectin. Results: Significant reduction in the autophagy biomarkers (p < 0.001) over the first 24 hours then began to increase but still not reach the normal level up to 72 hours after burn. Vitronectin level increased after burn infliction 1.5-fold after first hour, then up to four-fold after four hours and after that began to decline but still did not reach the normal level up to 72 hours. Conclusion: Autophagy biomarkers can be used as a forensic tool in determination of the time passed since burn infliction in living.



Author(s):  
М.В. Осиков ◽  
Е.В. Симонян ◽  
А.А. Агеева ◽  
А.И. Синицкий ◽  
Ю.И. Агеев

Разработка и патогенетическое обоснование новых подходов к локальной терапии термической травмы (ТТ) является актуальной и востребованной проблемой. В частности, представляет интерес разработка дермальных пленок (ДП), содержащих эндогенные регуляторы гомеостаза мультитропного действия. Цель исследования - оценка эффекта мелатонина (МТ) в составе оригинальной ДП на процессы репарации и содержание продуктов перекисного окисления липидов (ПОЛ) в коже очага повреждения при локальной термической травме. Методика. Эксперимент выполнен на 126 крысах-самцах Wistar. ТТ ІІІА степени площадью 3,5% моделировали погружением участка межлопаточной области кожи в очищенную воду с температурой 98-99 ºС на 12 с. МТ в составе ДП (0,005 г/г) на основе натрия карбоксиметилцеллюлозы наносили ежедневно после ТТ в течение 5 сут. На 5-е и 10-е сут после ТТ оценивали макроскопическую картину, площадь и глубину ожоговой раны, скорость ее эпителизации. Содержание продуктов ПОЛ в гомогенате кожи ожоговой раны определяли экстрационно-спектрофотометрическим методом в гептановой и изопропанольной фазах липидного экстракта. Результаты. Установлено, что накопление вторичных и конечных продуктов ПОЛ в гептановой и изопропанольной фазах липидного экстракта на 5-е и 10-е сут ассоциировано с площадью ожога. Применение оригинальной ДП с мелатонином приводит к снижению абсолютной и относительной площади ожога, увеличению скорости эпителизации ожоговой поверхности. На 5-е сут обнаружено снижение содержания вторичных и конечных продуктов ПОЛ в изопропанольной фазе, на 10-е сутки - снижение вторичных продуктов ПОЛ в гептановой фазе, конечных продуктов ПОЛ в изопропанольной фазе липидного экстракта. Заключение. Полученные результаты демонстрируют ускоряющий репарацию кожи в очаге ТТ эффект МТ в составе дермальной пленки за счет его ПОЛ-ограничивающего действия, расширяют представления о мультитропных эффектах МТ в организме и являются предпосылкой для применения ДП с МТ в клинической практике. Background. Development and pathogenetic justification of new approaches for local therapy of thermal trauma (TT) is a relevant and in-demand issue. Of special interest are dermal films (DF) containing endogenous pleiotropic regulators of homeostasis. Melatonin (MT) is one of such regulators that is suggested to be protective in TT. The aim of this study was to evaluate the effect of MT in the original DF on indexes of repair and concentration of lipid peroxidation (LPO) products in the injured skin after experimental TT. Methods. Experiments were performed on 126 Wistar male rats. Grade IIIA TT with an area of 3.5% was modeled by immersing a section of interscapular skin in purified water at a temperature of 98-99oC for 12 s. MT formulated into DF (0.005 g/g) based on sodium carboxymethylcellulose was applied after TT daily for 5 days. The macroscopic picture, area and depth of the burn wound, and the wound epithelization rate were evaluated on days 5 and 10 after TT. Concentration of LPO products in the injured skin homogenate was measured by extraction spectrophotometry in heptane and isopropanol phases of the lipid extract. Results. The accumulation of secondary and final LPO products in the heptane and isopropanol phases of the lipid extract on days 5 and 10 was associated with the burn area. The use of the original DF with MT resulted in a decrease in the absolute and relative areas of the burn and an increase in the rate of burn surface epithelialization. On day 5, a decrease in the content of secondary and final LPO products in the isopropanol phase was observed, and on day 10 decreases in secondary peroxidation products in the heptane phase and end LPO products in the isopropanol phase were detected. Conclusion. The results of this study demonstrated that MT formulated into DF accelerates skin repair in the TT focus due to its LPO-limiting effect, expands the understanding of MT pleiotropic effect, and represents a prerequisite for the clinical use of DF with MT.



2010 ◽  
Vol 01 (04) ◽  
pp. 279-288 ◽  
Author(s):  
Enas Hamed ◽  
Galia Awean ◽  
Nagla Abdel-Hafez ◽  
Youssef Hassan ◽  
Samer Zahran

AbstractBurn is an injury which is not only devastating for the patients but also puts a great burden on the society by consuming enormous health care resources. Despite improvements in burn wound care and treatment, understanding the role of key pro-/anti-inflammatory cytokines in pathogenesis of burn injury, sepsis and survival requires clarification. This work aimed at measuring plasma levels of the following cytokines: leptin, tumor necrosis factor (TNF)-α, interleukin (IL)-6, transforming growth factor (TGF)-α, and basic fibroblast growth factor (bFGF), procalcitonin (PCT) and C-reactive protein (CRP) in children with thermal burn (n = 42; 22 males and 20 females with an age range of two months up to 7 years) vs. sex and age matched healthy controls (n = 26; 12 males and 14 females with an age range of six months up to 8 years). The venous samples were drawn from patients at 2nd and 8th day postinjury. The relationship between measured plasma cytokines with length of hospital stay, total burn surface area (TBSA), sepsis, survival and complete blood count were investigated. The fatality rate was 28.6% (n = 12). At the 2nd day of postinjury, burn cases showed significantly higher values of WBC, CRP, PCT, TNF-α, IL-6, leptin, bFGF, and TGF-α than healthy controls. At the 8th day of postinjury, sepsis caused further significant elevations of WBC, CRP, PCT, TNF-α and IL-6 levels but significantly lower TGF-α levels than cases without sepsis. At the 8th day of postinjury, patients with larger TBSA (> 30%) demonstrated significantly higher values of WBC, CRP, PCT, TNF-α, and IL-6, but significantly lower levels of bFGF and TGF-α than cases with smaller TBSA (≤ 30%). At the 8th day of postinjury, non-survivors showed significantly higher levels of WBC, CRP, PCT, TNF-α and IL-6, but significantly lower levels of leptin, bFGF, and TGF-α than survivors. Correlation studies showed significant positive correlation between TBSA and each of IL-6, TNF-α and leptin levels. Except for bFGF and TGF-α levels that were reduced, the increases in other parameters (PCT, TNF-α and IL-6) were directly proportionate to the burn severity, sepsis and fatality. Both bFGF and TGF-α correlated with poor prognostic burn healing in severe septic cases. PCT and/or IL-6 are valuable biomarkers for early detection of burn infections. TGF-α, bFGF and leptin being immuno- and angiogenic modulators might reduce inflammation and post-burn immune dysfunction and enhance burn healing.



2012 ◽  
Vol 4 (2) ◽  
pp. 247-251
Author(s):  
Reema Pachnanda ◽  
Shiv Pal Singh

The present study was designed to investigate the adverse effect of arsenic on testicular tissue of Swiss albino male rats. Sodium arsenite was administered to adult male rats by gavage at the doses 1, 2 and 3 mg/kg body weight for 30 days. After the treatment, the testis were processed for histopathological observations. Sodium arsenite caused remarkable reduction in testicular weight (P<0.05), while the body weight of experimental animals were reduced but not significantly (P<0.05). Histological evaluation revealed dose-dependent, gradual destruction in histoarchitecture of testicular tissue. Sodium arsenite exposure caused complete arrest of spermatogenesis with disfigured seminiferous tubules in the testes .The lumens of the tubules were devoid of spermatids and were in places filled with cellular debris. The germinal epithelium was distorted. At places interstitial odema was also evident. Sertoli and Leydig cells were damaged. Along with structural alterations, fertility rate in experimental animals was significantly decreased at higher doses i.e. 2 and 3 mg/kg, as 100% infertility was observed. After withdrawal of the treatment over a period of 30 days, recovery was observed in low dose groups as few female rats became pregnant. The study concluded that exposure of arsenic causes testicular toxicity in male albino rat.



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