The study of biological objects using atomic force microscopy seems to be a promising direction, since it allows preserving their native structure. The use of atomic force microscopy to study nanostructural morphofunctional changes in cells of various tissues under normal conditions and pathologies can help accumulate new information about the pathogenesis of diseases, as well as become the foundation for the development of new approaches to their prevention and treatment.To study the possibilities of atomic force microscopy in studying pathomorphological changes in muscle, bone tissue and red blood cells in diabetes mellitus. The study demonstrated the possibility of using atomic force microscopy to study changes in body tissue under pathological conditions.

Objective: Experimental study of the damaging effect of cytostatics on the oral mucosa, disruption of the protective mechanisms of the mucous membrane, as well as assessment of the reversibility of these changes. Materials and methods: Using histological, morphometric, and quantitative histochemical methods, the mucous membrane of the tongue was studied on 40 mature white outbred mice after intraperitoneal administration of the cytostatic cyclophosphamide (CP) at a dose of 400 mg/kg for 5 days (20 animals). Animals in the control group (20 mice) were injected with isotonic sodium chloride solution at the same frequency. The material was obtained 24 hours and 20 days after the last injection of the drug. The condition of the oral mucosa (OM) on the ventral surface of the tongue was assessed. Results: Exposure to CP led to damage to all layers of the mucous membrane: the processes of proliferation and keratinization of the integumentary epithelium were disrupted, and synthetic processes in the epithelial cells of the small mixed salivary glands were disrupted. The number of granulocytes and mast cells decreased, and the relative volume of blood vessels in the connective tissue of the mucous membrane decreased. The changes were reversible. Conclusions: Cytostatic therapy leads to damage to the oral mucosa and disruption of its supraepithelial, epithelial and subepithelial protective mechanisms. There is a high degree of regeneration of the mucous membranes after the withdrawal of the cytostatic drug.

The introduction of new methodological approaches actualizes research that makes it possible to establish relationships between the elemental composition of hard dental tissues and microscopic analysis data. Using scanning electron microscopy, the nature of the location and size of enamel prisms was revealed, and a description of the structure of the initial and final enamel was given. The zone of the dentino-enamel junction, the location of the dentinal tubules and the composition of the dentin matrix were studied. It has been proven that the content of water, organic matter, sodium and potassium is higher in dentin, and the concentration of magnesium, phosphorus and calcium is higher in enamel.

Diabetes mellitus, as one of the most common socially significant diseases, has not lost its relevance for decades. Experimental models for studying this endocrinopathy are of scientific value not only in historical terms, but also greatly enrich modern ideas about various pathophysiological aspects of its course. Objective - to develop and test a new method for inducing diabetes mellitus (DM) with relative insulin deficiency for modeling gestational pathology. Materials and methods. The study was carried out on mature white female rats (30 individuals) of the Wistar line weighing 200-250 g. Using the method of pacreato-slenectomy described in the work, DM with relative insulin deficiency was induced in animals, after which ten female rats with hyperglycemia were transplanted to intact males. Results. Diabetes mellitus in laboratory animals was induced by removing the gastrosplenic part of the pancreas, which made it possible to reproduce the state of rather long-term hyperglycemia (up to 61 days of the experiment) with a high degree of both early and late survival. Female rats selected for the second stage of the experiment were able to become pregnant from intact males and carry offspring, while maintaining stable elevated blood glucose levels throughout the entire gestation period. Conclusion. The experimental model proposed by the authors can provide an opportunity for a long-term study of DM with relative insulin deficiency, as well as testing of new antidiabetic drugs, including against the background of pregnancy.

Objective: to detect possible causes of calcification of the placenta, identify patterns between excessive deposition of fibrinoid, calcium salts and various disorders in the mother-placenta-fetus system. Materials and methods. 6,089 afterbirths and placental tissue sections were examined, of which 193 specimens complying with the purpose of the study were further studied and divided into three groups (group of biological norm; afterbirths in which there is fibrinoid deposition without accompanying calcification; afterbirths with pronounced deposits of calcium salts). Each section of placental tissue was studied macro- and microscopically according to the method of A.P. Milovanov and A.I. Brusilovsky to reveal structural and functional changes. Statistical processing of the results was performed using methods of descriptive statistics, nonparametric methods: compilation of contingency tables, calculation of Pearson's χ2 test with Yates correction. Results. It was found that all the afterbirths studied had similar macroscopic characteristics. Microscopic examination revealed that calcification contributes to a more pronounced separation of the lobules of the maternal part of the placenta. Various inflammatory, alterative processes, as well as hemodynamic disorders in the placental tissue contribute to the deposition of calcium salts. Conclusion. The involutive processes observed in normal aging of the placenta become most pronounced in various pathological processes in the mother-fetus-placenta system. The study revealed a statistically significant relationship between the number of labor and the appearance of calcifications in the placental tissue. Large focal deposits of calcium salts were more often observed in women who gave birth not for the first time.

Background: The high incidence of diabetes mellitus in modern conditions dictates the need to search for new, including microscopic, opportunities to identify pathological changes in the course of various physiological processes. The aim of the study: To study the peculiarities of changes in the properties and parameters of erythrocytes in various types of diabetes mellitus (types 1 and 2) in women of mature and elderly age. Materials and methods: Venous blood of 55 women aged 30 to 70 years, divided into study groups (women with type DM, women with type II DM, control without endocrinopathy) was examined using atomic force and scanning probe microscopy. Results: The authors found an increase in the percentage of pathological forms of erythrocytes in patients with diabetes mellitus. An increase in the number of macrocytic forms of erythrocytes was observed not only when compared with the control group (DM I 12.20±2.14, DM II 10.00±1.20, control 4.40±0.15), but and when comparing two age groups of patients with type 2 DM (30-45 years old 10.00±1.20, 60-70 years old 13.20±1.90). The same tendency is typical for microcytes. Geriatric changes in the geometric parameters of erythrocytes in the group of patients with type 2 DM were characterized by an increase in the volume, as well as the maximum height of erythrocytes with increasing age of the patients. Conclusion: The use of atomic force microscopy in the study of the biophysical parameters of erythrocytes makes it possible to identify paОригинальная статья Original article Научные результаты биомедицинских исследований. 2021;7(4:400-409 Research Results in Biomedicine. 2021:7(4):400-409 401 tients from high-risk groups by determining such changes as an increase in the number of macrocytes, microcytes and pathologically altered forms of erythrocytes, a decrease in the average volume and area of erythrocytes.

Atomic force microscopy is not very popular in practical health care, therefore, its potential is not studied enough, for example, in obstetrics when studying the "mother-placenta-fetus" system. Our study summarizes the possibilities of using atomic force microscopy for detection of various circulatory disorders and vascular changes at the microscopic level in the uterus (endometrium and myometrium), placenta, and umbilical cord in the main variants of obstetric and endocrine pathology. For instance, in the case of endocrine pathologies, changes in the form of stasis, sludge, diapedesis, ischemia, destruction and separation of endotheliocytes in villous blood vessels were found in the mother. The oxygen content in erythrocytes also naturally decreased in pathologies; poikilo- and anisocytosis were observed.

Various uteroplacental blood flow disorders, their diagnosis, and correction, especially if the maternal obstetric history is burdened by endocrinopathy, still remain an urgent obstetric problem.OBJECTIVE:To study blood flow in the vessel-red blood cell segment and to search for changes in both the vessels of the villous tree of the placenta and uterus and the properties of blood corpuscles, by using modern microscopy methods (scanning probe microscopy, atomic force microscopy).MATERIAL AND METHODS:For macroscopic and microscopic examinations, fragments of the placenta, umbilical cord, and uterus were taken; venous blood samples were collected from patients without endocrinopathy, with gestational diabetes mellitus (GDM) or type 1 and type 2 diabetes mellitus (DM) prior to delivery. The data were statistically processed using both parametric (Student's t-test) and nonparametric (Fisher's exact test) methods.RESULTS:The authors found that microangiopathy and other metabolic changes in diabetes mellitus could lead to villous stromal sclerosis, the appearance of intermediate villi, changes in blood vessel shape, relief tortuosity, depth, and area, endothelial cell destruction, stasis, and thrombosis. In these endocrinopathies, there were also changes in maternal red blood cells: their polymorphism was observed and geometric parameters and deformability were impaired. In this case, cell shape impairment correlated with the severity of the above described vascular complications.CONCLUSION:The use of modern microscopy methods makes it possible to diagnose uteroplacental blood flow changes even in early pregnancy and to prevent the progression of uteroplacental blood flow disorders and placental insufficiency.

Новое в изучении клинико-морфологических аспектов при эндокринопатиях у беременных © Т.В.ПАВЛОВА 1 , В

The issue of effective and aesthetic wound healing, particularly, in mature and elderly individuals, remains relevant. Aim. To characterize the dynamics of skin wound healing in mature experimental rats and to compare the effectiveness of approved clinical injectable stimulants of regeneration (deproteinized hemoderivative of calf blood and collagen). Methods. This experimental blind study was conducted on 9-month-old Wistar male rats weighing 443.6±6.5 g. Square wounds of a standardized size (each side, 11 mm; wound depth, down to fascia; mean area, 123.5 ± 2.1 mm2) were inflicted on the rat back. Efficacy and mechanism of action of two injectable drugs were studied for 14 days. Effects of deproteinized hemoderivative of calf blood (DHB, Actovegin) and collagen (Collost) were compared with control wounds (saline treatment). The wound size and volume and the state of microcirculation were evaluated in dynamics. Results. The area of the wounds decreased by 9.1% at 7 days and 80% at 14 days in the control group; by 28.6% and 88%, respectively, in the DHB group; and by 33.3% and 91.6%, respectively, in the collagen group. In the control and DHA groups, the wound healing was predominantly horizontal while in the collagen group is was vertical. An increase in the wound area in the phase of acute inflammation during the first 3-5 days was observed only in the control and DHB groups; this increase was localized perpendicular to skin tension lines. This indicates the importance of preoperative planning not only for the lines and directions of surgical incisions and suturing but also for the subsequent treatment of wounds. Microcirculation in the skin immediately adjacent to the wound edge increased on day 3 only in the DHB group; in the collagen group, it increased on day 7; and on day 14, it was increased in all wounds of all groups with the greatest vascular effect being observed in the collagen group. Conclusion. Both regeneration stimulants were effective; however, the wound healing patterns and rates were different, which might be due to different mechanisms of action of the drugs. This fact should be taken into account in clinical practice in order to reduce the regeneration time as well as to reduce the size and aesthetics of postoperative scar.

Цель исследования - выявление особенностей динамики заживления кожной раны у экспериментальных крыс зрелого возраста; сравнение эффективности зарегистрированных и применяющихся в клинической практике инъекционных стимуляторов регенерации тканей: депротеинизированного гемодеривата крови телят и коллагена. Методика. Экспериментальное слепое исследование проведено на 9-месячных крысах самцах Wistar массой 443,6±6,5 г. Квадратную рану стандартного размера (каждая сторона 11 мм, глубина раны до фасции, площадь в среднем 123,5±2,1 мм2) наносили на поверхность спины животных. В течение 14 сут изучали эффективность и возможные механизмы действия на заживление кожных ран двух инъекционных лекарственных средств: депротеинизированного гемодеривата крови телят (ДГК, актовегин) и коллагена (Коллост) в сравнении с контролем (физиологический раствор натрия хлорида). В динамике оценивали изменение площади ран, их объема, состояния микрогемоциркуляции. Результаты. Площадь ран группы Контроль сократились к концу 7-х сут в 1,1 раза, 14-х - в 5,0 раза, группы ДГК - в 1,4 и 8,3 раза, группы Коллаген - в 1,5 и 11,9 раза соответственно. В группах Контроль и ДГК заживление ран проходило в горизонтальном направлении, в группе Коллаген - вертикальном. Увеличение площади ран в фазу острого воспаления (в первые 3-5 сут) было только в группах Контроль и ДГК, при которых оно было перпендикулярно линиям напряжения кожи. Это свидетельствует о значимости направлений хирургических разрезов и наложения швов не только при создании, но и при последующем лечении ран. Уровень микроциркуляции в коже, непосредственно примыкающей к краю раны, на 3-и сут увеличивался только в группе ДГК, на 7-е сут - в группе Коллаген, на 14-е сут - во всех ранах всех групп, но наибольшие изменения микрогемоперфузии были в группе Коллаген. Заключение. Установлена эффективность обоих стимуляторов регенерации. Вместе с тем скорость и характер заживления ран при их применении существенно различались, что связано с разными механизмами действия препаратов. Это необходимо учитывать в клинической практике при подборе препаратов для ускорения сроков регенерации раны, уменьшения размеров послеоперационного рубца и повышения его эстетичности. Aim. To characterize the dynamics of skin wound healing in mature experimental rats and to compare the effectiveness of approved clinical injectable stimulants of regeneration (deproteinized hemoderivative of calf blood and collagen). Methods. This experimental blind study was conducted on 9-month-old Wistar male rats weighing 443.6±6.5 g. Square wounds of a standardized size (each side, 11 mm; wound depth, down to fascia; mean area, 123.5 ± 2.1 mm2) were inflicted on the rat back. Efficacy and mechanism of action of two injectable drugs were studied for 14 days. Effects of deproteinized hemoderivative of calf blood (DHB, Actovegin) and collagen (Collost) were compared with control wounds (saline treatment). The wound size and volume and the state of microcirculation were evaluated in dynamics. Results. The area of the wounds decreased by 9.1% at 7 days and 80% at 14 days in the control group; by 28.6% and 88%, respectively, in the DHB group; and by 33.3% and 91.6%, respectively, in the collagen group. In the control and DHA groups, the wound healing was predominantly horizontal while in the collagen group is was vertical. An increase in the wound area in the phase of acute inflammation during the first 3-5 days was observed only in the control and DHB groups; this increase was localized perpendicular to skin tension lines. This indicates the importance of preoperative planning not only for the lines and directions of surgical incisions and suturing but also for the subsequent treatment of wounds. Microcirculation in the skin immediately adjacent to the wound edge increased on day 3 only in the DHB group; in the collagen group, it increased on day 7; and on day 14, it was increased in all wounds of all groups with the greatest vascular effect being observed in the collagen group. Conclusion. Both regeneration stimulants were effective; however, the wound healing patterns and rates were different, which might be due to different mechanisms of action of the drugs. This fact should be taken into account in clinical practice in order to reduce the regeneration time as well as to reduce the size and aesthetics of postoperative scar.

The aim of the research is study of histological features of the placen-ta’s and uterus’ structure, erythrocytes in the mother-placenta-fetus system during pregnancy, occurring against the background of thyroid diseases, diabetes (type I and gestational), moderate and severe preeclampsia.117 pregnant women were examined during the study: 65 were with pathology of the thyroid gland (TG); 27 with diabetes mellitus; 15 with preeclampsia and 10 were the control group. All women underwent ultrasound scanning on an Alokka-5500 apparatus, and each of them was determined the level of thyroid hormones and antibodies to the TG, as well as glucose. Tissue morphometric analy-sis was performed using the “Ntegra-Aura” device, atomic force image analysis using standard software NOVA and Image Analysis (“NT-MDT”). During the study of the placenta’s and myometrium’s vessels were identified different circulatory disorders. It was found that the area of the villi and vessels of the terminal placenta de-creased in the sequence: gestational diabetes mellitus (GDM), the state of euthyroidism in the mother with thyroid pathology, hypothyroidism, diffuse toxic goiter (DTG), preeclampsia and type I diabetes mellitus (DM I). A significant increase in deformed and prehemolytic forms of erythrocytes was revealed. Itwas found that in the mother-placenta-fetus structure with pathology of the TG, DM and preeclampsia, multiple structural changes in the villous tree's and myometrium's tissues are observed, as well as changes in anatomical and functional structure of erythrocytes, which together contribute to the progression of fetal hypoxia.

The aim of the study was to develop new approaches to the study of modern aspects of the clinical and morphological features of uterine blood flow in type I diabetes mellitus (DM) in the mother. Under our supervision, there were 40 pregnant women with type I DM. Clinical examination methods generally accepted in obstetric practice were used to assess the condition of the fetus, placenta, and blood flow in the fetal vessels and uterine arteries. Light microscopy, atomic force microscopy (AFM), and scanning electron microscopy (SEM) of 2 uterine fragments obtained during cesarean section were performed. Samples were examined using a FE1 Quanta 200 3D scanning microscope, analyzed, and morphometric and imaging were performed. Processing AFM images was formulated using the NOVA software (NT-MDT, Russian Federation). Results obtained allow us to judge the adverse effect of diabetes on the mother's body during pregnancy and childbirth. Among the complications of pregnancy, it should be noted: the state of preeclampsia (8%), fetoplacental insufficiency (88%), the threat of abortion (52%), polyhydramnios (25%).Morphologically, we revealed plethora of the endometrium. A change in the shape of the myometrium with the presence of fragmentary destructive changes in the myometrium, including focal necrosis and damage to the vascular system of the pregnant uterus. The paper substantiates the possibility of using scanning electron and atomic force microscopy as express methods for morphofunctional diagnosis of a woman's condition in pregnancy pathology.

The purpose of this research work was the creation of a safe and effective wound healing drug, in the multicomponent polysaccharide base of which the fibroblast growth factor is integrated in the optimal concentration. Materials and Methods: The present work was carried out in three stages. In Stage 1, the essential formula of a wound healing drug was created. In order to optimize the composition of the base formula, studies were conducted with various concentrations of structure-forming polysaccharides. In Stage 2, we performed technical tests of the gel composition of the medical product on hMSCs culture, and determined the optimal concentration of the growth factor, which was added to the base formula developed at Stage 1 in different concentrations. In Stage 3, an experimental safety study of the developed gel composition, including growth factors, was performed on laboratory animals. In an experimental safety study (subchronic toxicity) on a wound model, sexually mature male rats and Wistar females of the same age (8-9 months) were used. In all animals, 2 wounds of standard size were modeled on the back. Taking into account the weight and gender, rats were distributed evenly into two experimental groups. Group 1 included rats we treated with a polysaccharide gel based on polymers. This polysaccharide gel included rhFGF-b. Group 2 was used as control (without treatment). Results: The laboratory, histological and gravimetric studies of the internal organs and wounds of the animals in the experimental and control groups did not reveal any regular pathomorphological changes indicating the presence of the general toxicological properties of the studied products. This indicates the safety in vivo of the product being developed. Conclusion: The studies conducted in vitro and in vivo allow us to proceed with studies determining the effectiveness of the developed wound healing medical product on laboratory animals.