Polysulfides play an important role in regulating many physiological processes, making their study relevant in health and disease. This study aims to improve their evaluation method by determining alkaline-labile sulfane sulfur (ALSS) and to test it on various biological samples, as well as on models of hyperhomocysteinemia and oxidative/reductive stress. The ALSS assay is based on the photometric determination of H2S, which is generated from polysulfides under alkaline conditions. The limit of quantitative detection was 3.3 nmol, interday reproducibility was 5.7% (blood plasma), and the yields of H2S precipitation and product extraction were 97.8 and 89.7%, respectively. The specificity of the approach, determined with two different methods of sulfide destruction, was 80% on a model mixture of bovine albumin or blood plasma. A small but noticeable matrix effect was revealed. ALSS of control rats (n = 15) and human plasma (n = 10) was 133 (128; 135) and 177.6 (171.8; 183.6) μmol/g protein, respectively. Decreased ALSS levels were observed in rats with mild hyperhomocysteinemia (n = 14) and in patients with acute stroke (n = 20). Under 24h oxidative stress, ALSS decreased from 14.8 ± 1.3 to 10.4 ± 1.0 μmol/g protein (p = 0.0055) in the THP-1 culture, while 96h reductive stress caused it to increase from 10.9 ± 0.9 to 15.9 ± 2.0 μmol/g protein, p = 0.00034. The method presented in this paper allows for the assessment of ALSS in various biological matrices and has demonstrated sensitivity to oxidative stress in vitro and hyperhomocysteinemia in vivo. This makes it a promising indicator of sulfur metabolism disorders in pathological conditions.

Background. Neutrophil extracellular traps (NETs) are net-like structures that have been investigated in inflammatory diseases. However, the presence of NETs in infected persons without clinical symptoms has not been yet studied.Aim. To reveal NETs in healthy persons during and after the H1N1 influenza pandemic as well as to study the functional activity of NETs.Materials and methods. The study included two groups of volunteers (n = 10 in each group) aged 20–25 years. The first group of volunteers was examined in the absence of acute diseases during one month before the study and in the absence of chronic diseases in the medical history. Volunteers of the second group were in contact with patients with influenza, but did not get sick. The comparative study also included patients with acute inflammation in the abdominal cavity (appendicitis, cholecystitis, abscess; 12 patients) and 9 patients with non-specific ulcerative colitis. Neutrophils were isolated from the blood by the traditional method of Ficoll density centrifugation. The number, morphology, and functional activity of NETs were determined (by capture of Klebsiella pneumoniae). SYBR Green I-based fluorescence microscopy was used to visualize and quantify NETs.Results. In healthy volunteers who were not in contact with infected patients, spontaneous NETs formation did not occur. Neutrophils of persons who were in contact with infected patients spontaneously formed NETs. In this case the number of NETs reached 8.58 ± 0.51%, and the size of NETs amounted to 39.68 ± 3.52 µm. NETs effectively captured cells of the tested microorganism, which was accompanied by retraction of network fibers and transformation of the network structure into a cloud-like one, which retained 89.38 ± 5.86 microbial cells. For comparison, the NETs in patients with acute inflammation in the abdominal cavity captured and bound 20.2 ± 1 .67 microbial cells and with non-specific ulcerative colitis – 5.53 ± 0.34 cells.Conclusion. High binding capacity of NETs is a factor contributing to effective defense of the body against the development of an infectious disease with manifested clinical symptoms.

The antioxidant properties of cerium dioxide nanoparticles synthesized by the hydrothermal method are studied both by optical photometry and by EPR spectroscopy. It was found that with increasing intensity of irradiation of methylene blue with red light, the antioxidant activity of CeO2 nanoparticles improve. The greatest antioxidant activity is achieved at a concentration of CeO2 particles in solution from 5 mM to 10 mM. Increasing a concentration of CeO2 nanoparticles above 20 mM leads to deterioration in their antioxidant properties. It has been shown that in an alkaline medium the antioxidant activity of nanoparticles increases significantly, but with a decrease in pH to 3, CeO2 nanoparticles exhibit weak pro-oxidant properties due to more active generation of O2•− radicals.

Развитие инфекционных осложнений у больных с воспалительными заболеваниями определяется тем, насколько эффективную защитную реакцию нейтрофилы больного будут развивать при контакте с патогеном. Противоинфекционная устойчивость организма во многом определяется готовностью нейтрофилов формировать нейтрофильные сети, способные эффективно захватывать патогены. Цель исследования. Определение параметров нейтрофильных экстраклеточных ловушек (НЭЛ) у больных с воспалительными заболеваниями брюшной полости. Методика. Обследованы 28 прооперированных больных с различными формами абдоминального воспаления и 6 терапевтических больных с язвенным колитом, 6 пациентов обследованы при поступлении. Нейтрофилы выделяли, используя градиентное центрифугирование. Для подсчета НЭЛ использовали флюоресцентную микроскопию с красителем SYBR Green (Evrogen; Россия), специфично взаимодействующего с двухцепочечной ДНК. Функциональную активность НЭЛ определяли в тесте с захватом Klebsiella pneumoniae (ATCC 700603). Результаты. При исследовании больных в послеоперационном периоде обнаружили, что при остром аппендиците/абсцессе, остром холецистите и остром панкреатите/панкреонекрозе обнаруживаются близкие количества НЭЛ – 12,63±2,13%, 15,66±2,81% и 12,64±2,73%. В этих же группах больных регистрируются достоверные различия в размерах НЭЛ, которые достигают 46,82±6,70 мкм, 73,62±16,29 мкм и 96,84±27,17 мкм, соответственно. Увеличение размеров НЭЛ в этих группах больных объясняется присутствием, помимо сетевидных структур, дополнительно еще и нитевидных, волокнистых и вуалеобразных нейтрофильных внеклеточных структур. Группа больных с неспецифическим язвенным колитом, получающих консервативное лечение (группа сравнения), демонстрирует тенденцию к уменьшению размеров НЭЛ – до величины 31,96±14,75 мкм. Исследование функциональной активности НЭЛ показывает, что её снижение предшествует развитию септического состояния (на примере выявленного клинического случая). Заключение. Все, без исключения, исследованные нозологические формы воспаления в брюшной полости сопровождаются формированием НЭЛ в виде нейтрофильных сетей. Помимо нейтрофильных сетей воспаление индуцирует у части больных формирование НЭЛ в виде одиночных нитей, волокон и вуалей. Количественные параметры НЭЛ коррелируют с размерами и распространением очага воспаления на соседние анатомические области. На примере описанного клинического случая обнаружено резкое ослабление функциональной активности НЭЛ, которое предшествовало развитию септического осложнения. Прослеживается связь между исходно ослабленной функциональной активностью НЭЛ и развитием септического осложнения у пациента в послеоперационном периоде. The development of infectious complications in patients with inflammatory diseases is determined by how effective protective reaction the patient’s neutrophils will develop upon contact with the pathogen. The body’s anti-infective resistance is largely determined by the willingness of neutrophils to form neutrophilic networks capable of effectively capturing pathogens. Aim. Determination of parameters of neutrophil extracellular traps (NETs) in patients with inflammatory diseases of the abdominal cavity. Methods. 28 operated patients with various forms of abdominal inflammation and 6 therapeutic patients with ulcerative colitis were examined, 6 patients were examined upon admission. Neutrophils were isolated using gradient centrifugation. To calculate the NETs, fluorescence microscopy with the dye SYBR Green (Evrogen; Russia), which specifically interacts with double-stranded DNA, was used. The functional activity of NETs was determined in the Klebsiella pneumoniae capture test (ATCC 700603). Results. In the study of patients in the postoperative period, it was found that in acute appendicitis/abscess, acute cholecystitis and acute pancreatitis/pancreonecrosis, similar amounts of NETs are found – 12,63±2,13%, 15,66±2,81% and 12.64±2.73%. In the same groups of patients, significant differences in the size of the NETs are recorded, which reach 46.82±6.70 microns, 73.62±16.29 microns and 96.84±27.17 microns, respectively. The increase in the size of the NETs in these groups of patients is explained by the presence, in addition to network-like structures, additionally filamentous, fibrous and cloud-like neutrophil extracellular structures. A group of patients with nonspecific ulcerative colitis receiving conservative treatment (comparison group) shows a tendency to decrease the size of the NEL to a value of 31.96±14.75 microns. A study of the functional activity of NETs shows that its decrease precedes the development of a septic condition (using the example of an identified clinical case). Conclusion. All, without exception, the studied nosological forms of inflammation in the abdominal cavity are accompanied by the formation of NETs in the form of neutrophilic networks. In addition to neutrophilic networks, inflammation induces the formation of NETs in the form of single strands, fibers and clouds form in some patients. The quantitative parameters of the NETs correlate with the size and spread of the inflammatory focus to neighboring anatomical areas. On the example of the described clinical case, a sharp decrease in the functional activity of NETs was found, which preceded the development of a septic complication. There is a connection between the initially weakened functional activity of NETs and the development of septic complications in the patient in the postoperative period.

Objective: to analyze data on the synthesis and properties of cerium oxide nanoparticles, as well as the prospects of its application in regenerative medicine for wound healing. Methodology. World literature was reviewed using PubMed, SCOPUS, ResearchGate, CyberLeninck, and Elibrary databases, as well as manual searches for authors and reference lists. Key search terms were «cerium oxide» AND nano* AND (healing OR regeneration OR repair) AND wound». The timeline was from the date of publication through August 2023. Results. The final analysis included 59 sources containing information on the synthesis and size of nanoparticles (and/or other physicochemical characteristics), methodology and results of in vivo and in vitro studies on the efficacy and/or safety of nanoceria for wound regeneration. It is shown that despite the progressive growth of research interest over the last 15 years, the actual use of nanoceria in practical medicine is still not widespread. This is due to a wide variety of non-standardized synthesis methods and conditions, resulting in the variability of physicochemical parameters of nanoparticles (size, form), thereby affecting the safety and biomedical efficacy of nanoceria. Regeneration mechanisms, including the antioxidant-prooxidant, anti-inflammatory and antimicrobial effects of nanoceria, which contribute to accelerated wound healing, are discussed. The severity of the regenerative effects depends on the method and conditions of synthesis, hence the resulting physicochemical characteristics of the nanoparticles. Therefore, after each batch, newly synthesized nanoceria needs physicochemical and biomedical experimental tests. Conclusion. Nanoceria has great potential in tissue engineering for regenerative medicine, particularly for healing of various kinds of wounds. Having developed a technology for standardized synthesis for effective and safe nanoceria (of the right form and size) on a production scale, it can be introduced in medicine, possibly improving the outcomes of treatment of many diseases and pathologies. The authors present conclusions on the results of the study of nanoceria for accelerating qualitative regeneration and the requirements for nanoparticles obtained during synthesis.

The purpose of this study was to investigate the antimicrobial activity of citrate-stabilized sols of cerium oxide nanoparticles at different concentrations via different microbiological methods and to compare the effect with the peroxidase activity of nanoceria for the subsequent development of a regeneration-stimulating medical and/or veterinary wound-healing product providing new types of antimicrobial action. The object of this study was cerium oxide nanoparticles synthesized from aqueous solutions of cerium (III) nitrate hexahydrate and citric acid (the size of the nanoparticles was 3–5 nm, and their aggregates were 60–130 nm). Nanoceria oxide sols with a wide range of concentrations (10−1–10−6 M) as well as powder (the dry substance) were used. Both bacterial and fungal strains (Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris, Candida albicans, Aspergillus brasielensis) were used for the microbiological studies. The antimicrobial activity of nanoceria was investigated across a wide range of concentrations using three methods sequentially; the antimicrobial activity was studied by examining diffusion into agar, the serial dilution method was used to detect the minimum inhibitory and bactericidal concentrations, and, finally, gas chromatography with mass-selective detection was performed to study the inhibition of E. coli’s growth. To study the redox activity of different concentrations of nanocerium, we studied the intensity of chemiluminescence in the oxidation reaction of luminol in the presence of hydrogen peroxide. As a result of this study’s use of the agar diffusion and serial dilution methods followed by sowing, no significant evidence of antimicrobial activity was found. At the same time, in the current study of antimicrobial activity against E. coli strains using gas chromatography with mass spectrometry, the ability of nanoceria to significantly inhibit the growth and reproduction of microorganisms after 24 h and, in particular, after 48 h of incubation at a wide range of concentrations, 10−2–10−5 M (48–95% reduction in the number of microbes with a significant dose-dependent effect) was determined as the optimum concentration. A reliable redox activity of nanoceria coated with citrate was established, increasing in proportion to the concentration, confirming the oxidative mechanism of the action of nanoceria. Thus, nanoceria have a dose-dependent bacteriostatic effect, which is most pronounced at concentrations of 10−2–10−3 M. Unlike the effects of classical antiseptics, the effect was manifested from 2 days and increased during the observation. To study the antimicrobial activity of nanomaterials, it is advisable not to use classical qualitative and semi-quantitative methods; rather, the employment of more accurate quantitative methods is advised, in particular, gas chromatography–mass spectrometry, during several days of incubation.

Background. Neutrophil extracellular traps (NETs) are net-like structures that have been investigated in inflammatory diseases. However, the presence of NETs in infected persons without clinical symptoms has not been yet studied. Aim. To reveal NETs in healthy persons during and after the H1N1 influenza pandemic as well as to study the functional activity of NETs. Materials and methods. The study included two groups of volunteers (n = 10 in each group) aged 20-25 years. The first group of volunteers was examined in the absence of acute diseases during one month before the study and in the absence of chronic diseases in the medical history. Volunteers of the second group were in contact with patients with influenza, but did not get sick. The comparative study also included patients with acute inflammation in the abdominal cavity (appendicitis, cholecystitis, abscess; 12 patients) and 9 patients with non-specific ulcerative colitis. Neutrophils were isolated from the blood by the traditional method of Ficoll density centrifugation. The number, morphology, and functional activity of NETs were determined (by capture of Klebsiella pneumoniae). SYBR Green I-based fluorescence microscopy was used to visualize and quantify NETs. Results. In healthy volunteers who were not in contact with infected patients, spontaneous NETs formation did not occur. Neutrophils of persons who were in contact with infected patients spontaneously formed NETs. In this case the number of NETs reached 8.58 +/- 0.51%, and the size of NETs amounted to 39.68 +/- 3.52 mu m. NETs effectively captured cells of the tested microorganism, which was accompanied by retraction of network fibers and transformation of the network structure into a cloud-like one, which retained 89.38 +/- 5.86 microbial cells. For comparison, the NETs in patients with acute inflammation in the abdominal cavity captured and bound 20.2 +/- 1 .67 microbial cells and with non-specific ulcerative colitis - 5.53 +/- 0.34 cells. Conclusion. High binding capacity of NETs is a factor contributing to effective defense of the body against the development of an infectious disease with manifested clinical symptoms.

We developed a method of sensitive capillary electrophoresis using UV detection for the determination of certain free aminothiols (reduced cysteinylglycine (rCysGly), cysteine (rCys), glutathione (rGln), and cystine (CysS) in human blood plasma. The reduced thiols were derivatized with N-ethylmaleimide. The plasma was purified from proteins via ultrafiltration. Electrophoretic separation was performed using 115 mM Na phosphate with 7.5% (v/v) polyethylene glycol 600, pH 2.3. The in-capillary concentration of the analytes was achieved with a pH gradient created via the preinjection of triethanolamine and postinjection of phosphoric acid. The separation was carried out using a silica capillary (50 µm i.d.; total/effective separation length 42/35 cm) at a 25 kV voltage. The total analysis/regeneration time was 18 min. The quantification limits varied from 1.3 µM (rCysGly) to 5.4 µM (CysS). The accuracy was 95%-99%, and the repeatability and reproducibility were approximately 1.8%-3.8% and 1.9%-5.0%, respectively. An analysis of plasma samples from healthy volunteers (N = 41) showed that the mean levels of rCysGly, rCys, rGln, and CysS were 1.64, 10.6, 2.58, and 46.2 µM, respectively.

We examined standard clinical and laboratory biochemical parameters, as well as the levels of aminothiols in the blood and urine (homocysteine (Hcy), cysteine (Cys), S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH)) via capillary electrophoresis in patients with CKD at stages II–V. Patient outcomes were assessed after five years. To complete forecasting, correlation and ROC analysis were performed. It was found that the levels of Cys and Hcy in blood plasma were earlier markers of CKD starting from stage II, while the levels of SAM and SAM/SAH in urine made it possible to differentiate between CKD at stages II and III. Blood plasma Hcy and urinary SAM and SAM/SAH correlated with mortality, but plasma Hcy concentrations were more significant. Thus, plasma Hcy, urine SAM, and SAM/SAH can be considered to be potential diagnostic and prognostic markers in patients with CKD.

We studied the neutrophils and monocytes obtained from 37 patients with various inflammatory diseases such as psoriasis, acute infectious process in the abdominal cavity (acute appendicitis/abscess of the abdominal cavity, and acute cholecystitis), acute pancreatitis, and post-COVID syndrome after mild COVID infection. The number and the morphological structure of neutrophil extracellular traps (NET) as well as the effect of IgG on NET were examined. NET were visualized and counted by fluorescence microscopy with fluorescent dye SYBR Green. All the studied types of inflammation were accompanied by spontaneous formation of NET. After application of IgG, the number of NET doubled, their size increased, and transformation of net-like traps into the cloud forms was observed. The clouds form structure of the network is not capable of capturing pathogens with subsequent retraction, the products of its enzymatic degradation can be the factors of secondary alteration. The study results demonstrate a previously unknown mechanism of infection resistance.

OBJECTIVE:The purpose of the study was to examine the effect of cilostazol on isoforms of targets of phosphodiesterase 3 and phosphodiesterase 5 by means of molecular modeling. MATERIAL AND METHODS:Inhibition of phosphodiesterases (PDEs) was studied by means of molecular docking using software Maestro Schrodinger Inc. The structure of all mentioned proteins was taken from the Protein Data Bank (PDB): 7L27-PDE3A, 1JOS-PDE3B, 4MD6-PDE5. The structures of cilostazol and vardenafil ligands were obtained from the PubChem database. The Glide Docking SP mode was used to determine the best pose of the ligand in the supramolecular complex PDE-ligand with the function of minimal assessment of energy. The structures of PDEs were taken from protein database. Each molecule of PDE was prepared before docking using the Protein Preparation Wizard module. The ligand geometry was optimized using the force field MMF4 in the software package of molecular dynamics GROMACS. The active center of enzymes was considered rigid, with the change of torsion angles being allowed for cilostazol. The results were visualized using the function Ligand Interaction in the Maestro module. The inhibition constant (Ki) was obtained from the binding energy (ΔG) using the formula: Ki=exp (ΔG/RT), where R is the universal gas constant (1.985 × 103 kcal · mol 1 · K 1) and T is the temperature (298.15 К). RESULTS:Cilostazol possesses higher affinity to isoform of PDE3A (Ki=54 nM) as compared with PDE3B (Ki=1.13 μM) based on the findings of the performed molecular docking. Also, cilostazol can inhibit PDE5 in therapeutic doses (Ki=10 μM). CONCLUSION:Cilostazol-mediated inhibition of the isoform PDE3A to a greater degree than PDE3B explains a low incidence of side effects from the side of PDE3B inhibition in clinical practice. Cilostazol may also inhibit PDE5 at therapeutic doses (Ki=10 μM), in which connection it may have an additional therapeutic effect in treatment of intermittent claudication, diabetes mellitus and its complications, especially neuropathy.

Diabetes mellitus is a top priority non-communicable disease with a high risk of developing complications with distinctive features. Diagnosis of the stage and types of damage to the foot functions in patients with diabetes is a complex task. In presented clinical case, we used several tests to identify the expression of diabetic angiopathy: pulse examination of the main arteries, level of glycated hemoglobin (HbA1c), ultrasound of the low extremities arteries, transcutaneous pulse oximetry, and oxygen partial pressure examination in tissues; carboxyangiography or contrast angiography were performed for all patients. The admitted patient was diagnosed with a diabetic foot syndrome. Based on the partial pressure and oxygen saturation in the tissues of the right lower limb, it was decided to minimize the extremities height amputation, thus preserving the knee joint.

A new approach has been developed for the direct determination of reduced (glutathione [GSH]) and oxidized (glutathione disulfide [GSSG]) GSH in whole blood by means of capillary electrophoresis. Its features include GSH-stabilizing sample preparation, the use of an internal standard, and pH-mediated stacking. Blood stabilized with acid citrate and K3 EDTA was treated with acetonitrile with N-ethylmaleimide, and then the analytes were extracted with diethyl ether. The total analysis time was 8 min using a 50-µm (i.d.) by 32.5-cm (eff. length) silica capillary. The background electrolyte was 0.075-M citrate Na pH 5.8 with 200-µM cetyltrimethylammonium bromide and 5-µM sodium dodecyl sulfate, and the separation voltage was -14 kV. The quantification limit (S/N = 15) of the method was 1.5 µM for GSSG. The accuracy levels of GSH and GSSG analysis were 104% and 103%, respectively, and between-run precision levels were 2.6% and 3.2%, respectively. Analysis of blood samples from healthy volunteers (N = 24) showed that the levels of GSH and GSSG and the GSH/GSSG ratio in the whole blood were 1.05 ± 0.14 mM, 3.9 ± 1.25 µM, and 256 ± 94, respectively. Thus, the presented approach can be used in clinical and laboratory practice.

The urgency of the problem of wound healing is not in doubt, given the global trend of an increase in the number of operations and injuries with skin damage, as well as the lack of universal means of treating wounds.STUDY OBJECTIVE:To compare the effectiveness of the developed drugs, smart polymeric nano-drug with cerium oxide nanoparticles (SPN), and smart polymeric nano-drug in combination with mesenchymal stem cells (SPN + SC) on the healing process of skin wounds.MATERIAL AND METHODS:An experimental study was carried out using Wistar rats of post-reproductive age, which had dermis and epidermis removed on their backs. There were four groups of wounds in total: control, treatment with mesenchymal stem cells (SC), SPN, and SPN + SC.RESULTS:A positive therapeutic effect of polymeric drugs on the dynamics of wound area reduction was established, which was most typical for wounds of the SPN group and, particularly, the SPN + SC group. On the third day, an anti-inflammatory effect was revealed in the SC and the SPN + SC groups in particular, which was expressed in a reduced leukocyte infiltration and an increase in the level of microcirculation during this period. The fastest transition from the phase of exudation to proliferation was recorded in the SPN and SPN + SC groups. Histologically, these groups showed faster regeneration, including the epithelialization of wounds.CONCLUSION:The results obtained in the course of the study open up possibilities for the development of fundamentally new, highly effective wound healing agents.

Purpose: To investigate the comparative effectiveness of certain biological injectable stimulants for the healing of skin wounds and criteria for its assessment. Materials and Methods: A comparative study of the effectiveness of mesenchymal stem cells (SC group), collagen (Collagen group), and deproteinized calf blood hemoderivative (DCBH group) was carried out using an acute wound model. Control wounds were injected with isotonic sodium chloride solution (Control group). A total of four groups (28 wounds per group) were included in the study. Aged male Wistar rats were used as experimental animals. A dynamic assessment of the wound areas and edges, microvasculature assessment via laser Doppler flowmetry, histological and morphometric analyses to determine the quantitative and qualitative fibroblasts composition, as well as the degree of newly synthesized collagen maturity, was conducted on days 0, 3, 7, and 14. Results: The administration of SCs provided a rapid but short-lasting effect, whereas the administration of collagen resulted in a delayed but long-lasting wound-healing effect. DCBH resulted in little to no effect. An increase in the perfusion volume of the wound edges accelerated the regeneration process, while the level of microcirculation did not affect the number and activity of fibroblasts. The wound healing acceleration, as well as the new collagen and stratified epithelium formation and maturation, was associated with the presence of a sufficient pool of mature and active fibroblasts in the wound, and not with the number of fibroblasts. Conclusion: The present results clarify the action mechanisms of the studied drugs. In addition, the application purposes and different effects of each drug on the different wound healing phases were demonstrated. An assumption on the multi-component treatment advisability under the wound condition objective assessment possibility was made. Findings from this study may assist clinicians in making an informed transition to personalized wound management and achieve better clinical outcomes.

1Department of Human Pathology, I.M. Sechenov First Moscow State Medical University (Sechenov University), Moscow 119991, Russia; 2Department of Plastic and Reconstructive Surgery, Cosmetology and Cell Technologies, Pirogov Russian National Research Medical University, Moscow 117997, Russia; 3Department of Pathophysiology, I.M. Sechenov First Moscow State Medical University (Sechenov University), Moscow 119991, Russia; 4Department of Hospital Surgery No1, Pirogov Russian National Research Medical University, Moscow 117997, Russia Purpose: To investigate the comparative effectiveness of certain biological injectable stimulants for the healing of skin wounds and criteria for its assessment. Materials and Methods: A comparative study of the effectiveness of mesenchymal stem cells (SC group), collagen (Collagen group), and deproteinized calf blood hemoderivative (DCBH group) was carried out using an acute wound model. Control wounds were injected with isotonic sodium chloride solution (Control group). A total of four groups (28 wounds per group) were included in the study. Aged male Wistar rats were used as experimental animals. A dynamic assessment of the wound areas and edges, microvasculature assessment via laser Doppler flowmetry, histological and morphometric analyses to determine the quantitative and qualitative fibroblasts composition, as well as the degree of newly synthesized collagen maturity, was conducted on days 0, 3, 7, and 14. Results: The administration of SCs provided a rapid but short-lasting effect, whereas the administration of collagen resulted in a delayed but long-lasting wound-healing effect. DCBH resulted in little to no effect. An increase in the perfusion volume of the wound edges accelerated the regeneration process, while the level of microcirculation did not affect the number and activity of fibroblasts. The wound healing acceleration, as well as the new collagen and stratified epithelium formation and maturation, was associated with the presence of a sufficient pool of mature and active fibroblasts in the wound, and not with the number of fibroblasts. Conclusion: The present results clarify the action mechanisms of the studied drugs. In addition, the application purposes and different effects of each drug on the different wound healing phases were demonstrated. An assumption on the multi-component treatment advisability under the wound condition objective assessment possibility was made. Findings from this study may assist clinicians in making an informed transition to personalized wound management and achieve better clinical outcomes.

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.