Highlights

There is an association of varying strength and significance between serum endocan and endoglin levels and a number of clinical and instrumental characteristics of patients with coronary artery disease.

Abstract

Aim. To assess the relationship between serum endocan and endoglin levels with clinical and instrumental characteristics of patients with coronary artery disease (CAD).

Methods. The study included 264 people (161 men and 103 women), of whom 220 were patients with CAD. The patients underwent anthropometric measurements; coronary angiography; echocardiographic examination; duplex ultrasound scanning of the extracranial sections of the brachiocephalic arteries. We studied serum endocan (ng/mL), endoglin (ng/mL) and glucose (mmol/L) levels. The patients were divided into groups according to the SYNTAX score: group 1 – ≤ 22 points (124 people); group 2 – 23–32 points (53 people); group 3 – > 33 points (43 people). The control group was represented by healthy volunteers (44 people). All groups were comparable in age and gender. The differences were considered statistically significant at p < 0.05.

Results. We found an association between endocan level and severity of coronary artery (CA) lesions (r = 0.32, p < 0.001). In the group 1, the median endocan level was 14.40 ng/mL [10.19; 19.91], in the group 2 it was 20.31 ng/mL [12.75; 24.12], in the group 3 it was 32.10 ng/mL [22.12; 38.21], and in the control group it was 5.97 ng/mL [4.38; 8.25] (p < 0.0001). While studying serum endoglin levels, we did not find any statistical differences between the patient groups (p > 0.05). However, we found an association of varying strength and significance between the serum endocan and endoglin levels and a number of clinical and instrumental characteristics of patients. The results showed statistically significant differences in endocan levels between the groups of patients with multivessel disease (p < 0.01), angina pectoris (p < 0.0001), myocardial infarction in history (p < 0.0001) and obesity (p < 0.05) compared with patients without these diseases. While studying serum endoglin levels, we noted statistically significant differences in endoglin levels between the groups of patients with multivessel disease and patients with CA lesions only (p < 0.0001), as well as between patients with persistent atrial fibrillation and patients with sinus rhythm (p < 0.05).

Conclusion. The data obtained demonstrate the potential role of endocan and endoglin as promising biomarkers for diagnosing the severity of endothelial dysfunction in patients with CAD.

Highlights

The effectiveness of cognitive rehabilitation (the absence of the postoperative cognitive dysfunction (POCD) is associated with the emotional regulation of behavior and frequency-spatial organization of brain activity.

The positive outcome of cognitive rehabilitation is promoted by «pre-tuning» background theta activity associated with the components of emotional intelligence: «attention to emotions» and «use of emotions in decision making». The distinctive feature of the group with unsuccessful rehabilitation is the link between empathy and high-frequency beta 2 rhythm.

Aim. To study the relationship between the components of emotional intelligence (EI) and electrical activity of the brain (EEG), reflecting the effects of cognitive rehabilitation using multitasking training in cardiac surgery patients.

Methods. The prospective cohort study included 83 patients who had undergone direct myocardial revascularization, 42 of whom underwent multitasking training in the early postoperative period. Physical, laboratory, and extended neurophysiological examinations were performed before cardiac surgery and 11–12 days after the intervention.

Results. The effectiveness of cognitive rehabilitation (absence of postoperative cognitive dysfunction (POCD)) is related to peculiarities of emotional regulation of behavior, reflected in the ratio of components of EI and frequency-spatial organization of brain activity. The group of patients with positive cognitive rehabilitation (absence of POCD) is characterized by «pre-tuning» background theta activity associated with the components of emotional intelligence: «attention to emotions» and «use of emotions in decision making». The distinctive feature of the group with unsuccessful rehabilitation is the link between empathy and high-frequency beta 2 rhythm.

Conclusion. Regional features of detected coupling patterns of EI and brain activity suggest that the effectiveness of cognitive rehabilitation using multitasking training is determined by the degree of preservation of cortical control functions during the development of cardiac artery disease, reflected in the peculiarities of emotional regulation of behavior of the patients.

Highlights

• Telomere length in patients with coronary artery disease is shorter compared to the telomere length of healthy individuals.
• Telomeric regions of DNA in patients with coronary artery disease do not recover over time.
• Telomere length in atherogenesis negatively correlates with markers of cardiovascular diseases and inflammatory process.

Aim. To assess the impact of changes in the length of telomeric regions of chromosomes on the course of coronary artery disease caused by coronary artery atherosclerosis in the population of residents of a coal-mining region.

Methods. The study included 60 patients with coronary artery disease (before surgery and five years after) and 52 healthy participants. We isolated the DNA using the standard phenol-chloroform extraction method. We used the quantitative PCR method to measure the relative length of leukocyte telomeres and analyzed biochemical parameters using standard methods, selecting cytokine proteins as markers of the inflammatory process.

Results. Telomeres in healthy participants were seven times longer compared to patients with coronary artery disease. At the same time, the length of telomeric regions of DNA did not differ in patients before surgery and after 5 years of rehabilitation. We used ROC analysis to determine the effectiveness of measuring telomeres as a marker of atherosclerosis. The area under the ROC curve was 0.998 ± 0.002. We found an inverse correlation between the telomere length and such parameters as the total body sodium, triglycerides and high-density lipoproteins. We noted a significant inverse correlation between such indicators of the lipid profile as triglycerides and high-density lipoproteins only in patients before surgery. The study results revealed direct and inverse dependence of the length of telomeres and cytokines such as IL-33 and IL-10, respectively.

Conclusion. Supposedly, inflammatory processes and oxidative stress, complementing each other, are the causes of irreparable damage to telomeres, accelerating the aging process and leading to irreversible consequences in atherogenesis.

Highlights

Contemporary algorithms for optical coherence tomography (OCT)-guided percutaneous coronary intervention (PCI) are easy to learn and understand, however, these algorithms are mostly declarative, and not substantiated in practice. In particular, using the mean reference lumen diameter may result in underestimation of the optimal stent size in relation to the degree of stenosis, i.e., the difference between the minimum and maximum diameters (lumen asymmetry), whereas choosing a stent based on the mean lumen diameter leads to stent underexpansion.

Aim. To assess whether choosing a stent size based on the mean lumen diameter of the reference segment of the artery may result in its underexpansion.

Methods. The prospective cohort study included patients with chronic coronary artery disease and complex lesion anatomy who underwent high-risk OCT-guided PCI. Based on the OCT results, the distal and proximal reference segments were determined, in which the minimum and maximum diameters were measured, followed by calculating the mean lumen diameter, which was further used for stent selection. Based on the results of repeated OCT after stent implantation, the degree of stent expansion was assessed. The expected relationship between the degree of stent expansion and the baseline lumen asymmetry was assessed by correlation analysis. Linear regression was used to determine the asymmetry threshold corresponding to clinically relevant stent expansion of 80%.

Results. Overall, 24 patients were enrolled, 3 of them were excluded because of inability to perform OCT without lesion predilatation. On average, the maximum and minimum diameters of the reference lumen according to OCT were 2.87 ± 0.66 mm and 1.85 ± 0.44 mm, respectively. The ratio between minimum and maximum diameter was 0.35 ± 0.09. The mean diameter was 2.30 ± 0.51 mm, the difference between the maximum and mean diameters was 0.61 ± 0.28 mm. The criterion of stent expansion ≥ 80% was achieved in 6 cases (28.6%). The correlation analysis has shown a statistically significant inversely proportional dependence of the relative stent expansion on the baseline lumen asymmetry (the difference between the maximum and minimum diameters (r = –0.41, p = 0.032), while clinically relevant stent expansion ≥ 80% was observed with a relative difference in diameters of less than 30%

Conclusion. In case of significant lumen asymmetry choosing a stent diameter based on the mean lumen diameter is associated with stent uderexpansion.

Highlights

• Low-density lipoprotein cholesterol (LDL)-containing circulating immune complexes (LDL-CIC) reflect impaired lipid metabolism and systemic inflammation.
• Serum level of LDL-CIC is sensitive and specific diagnostic and prognostic marker of carotid atherosclerosis.
• The combination of age and serum LDL-CIC is particularly valuable for the prognosis of carotid atherosclerosis.

Aim. Recent studies suggested that increased level of low-density lipoprotein cholesterol (LDL)-containing circulating immune complexes (LDL-CIC) might have high diagnostic value in regards to clinical and asymptomatic atherosclerosis. The aim was to study prognostic significance of LDL-CIC.

Methods. The two-year study included 98 men. Each included patient underwent the following studies: carotid intima-media thickness, total cholesterol, low- and high-density lipoprotein cholesterol, triglycerides, and LDL-CIC.

Results. Among these parameters, low-density lipoprotein cholesterol and LDL-CIC were significantly associated with severity and prognosis of carotid atherosclerosis. LDL-CIC has the highest sensitivity and specificity in diagnostics of carotid atherosclerosis when combined with age. Other parameters do not affect the prognostic significance of LDL-CIC.

Conclusion. The results suggest that LDL-CIC can be used for diagnostics and prognosis of atherosclerosis.

Highlights:

• Epoxy-treated xenopericardium modified with polyvinyl alcohol has increased resistance to calcification and proteolytic degradation in vitro.
• Cyclic loads negatively affect fatigue life of studied material, moreover, this process is accompanied by deterioration of its anti-calcium and anti-enzyme properties. Additional studies aimed at improving the wear resistance of the hydrogel in the biological material are required.

Aim. To study the resistance of epoxy-treated bovine pericardium modified with polyvinyl alcohol to calcification and proteolytic degradation in collagenase after exposure to cyclic loading.

Material and methods. The epoxy-treated patches made with xenopericardium were treated with polyvinyl alcohol according to the original method, after that they were subjected to cyclic loading (70 million cycles) using the HiCycle wear tester system. Visualization of the surface and internal structure of the biomaterial was performed by scanning electron microscopy. The resistance to calcification was assessed by incubating the samples in a solution saturated with calcium ions and phosphate ions for 3 and 6 weeks, followed by quantitative measurement of the calcium by spectrophotometry. The susceptibility of the samples to proteolytic degradation was determined by weight loss after incubation in a solution of clostridial collagenase for 24 hours. The control group consisted of patches of unmodified epoxy-treated bovine pericardium.

Results. After cyclic loading, patches of xenopericardium modified with polyvinyl alcohol showed signs of fatigue (cracks on the surface and large cavities in the internal structure). Although cyclic loading significantly worsened the resistance of the modified tissue to calcification and proteolytic degradation (by 3 and 5 times, respectively, relative to the values obtained for patches stored under static conditions), patches from this group showed better results compared to unmodified patches. Thus, they contained 1.5 to 2 times less calcium after 3 and 6 weeks of incubation in a calcium-saturated solution. Mass loss after incubation in collagenase was 1.5 times lower for patches of modified biomaterial undergoing cyclic loading compared to unmodified patches.

Conclusion. The proposed modification method of the epoxy-treated xenopericardium with polyvinyl alcohol increases the resistance of biological tissue to calcification and proteolytic degradation. Although cyclic loading negatively affects the protective properties of the polymer coating over time, this type of modification can potentially slow down the degeneration of biomaterial used in manufacturing of bioprosthetic heart valves.

Highlights

• Micro-computed tomography allows specialists to qualitatively and quantitatively assess the structure of calcified areas of explanted bioprosthetic heart valves.
• This method enables the evaluation of changes in the structure of the bioprosthesis that have occurred during its prolonged operation.

Aim of the study. To evaluate the potential of high-resolution tomography for the study of mitral valve bioprostheses of different designs explanted due to dysfunction and various calcification patterns (microcalcification and macrocalcification).

Methods. Single samples of the «UniLine» and «PeriCor» bioprostheses were the objects of study, they were explanted due to dysfunction developed after 76 and 87 months of operation in recipients. The peculiarities of calcification localization in the structure of bioprostheses were studied using high-resolution tomography followed by reconstruction of volumetric images and quantitative analysis of radiodense areas. Moreover, we used light microscopy with Alizarin Red S staining to detect calcifications.

Results. The study results showed that the nature of the distribution and volume of calcinates significantly differ between the studied samples: for the «UniLine» bioprosthesis, the affected areas were located in the leaflet material and constituted 21.1% of the total biological tissue volume; for the «PeriCor» bioprosthesis, calcifications were diffusely distributed in small structural formations, accounting for a total of 5.1% of the biological material, primary localized on the «auxiliary» structures of the prosthesis – the covering made of porcine or calf xenopericardium. In addition, high-resolution tomography allowed us to determine the degree of deformation of the «UniLine» bioprosthesis frame, with the posts deviating inward by 1.1–1.4 mm.

Conclusion. The possibility of using computed microtomography for qualitative and quantitative assessment of calcified xeno-pericardial and xeno-aortal bioprostheses has been demonstrated. However, this method is limited in its ability to detect macrocalcification within the leaflet thickness.

Highlights

The study involves experiments with titanium substrates with various composite coatings. The aim of the study was to assess different methods of diagnostics of artificial implant materials and evaluate the effect of coatings on osteogenic differentiation in interstitial cells of the human aortic valve.

Background. Currently, the only medical treatment for aortic stenosis is aortic valve replacement. At the same time, calcific aortic stenosis is the third most prevalent cardiovascular disease that affects up to 10% of the population by the age of 80. In this regard, it is important to develop materials for aortic valve prostheses that would be highly biocompatible, chemically stable, non-thrombogenic, non-immunogenic and non-osteogenic.

Aim. To assess the impact of composite coatings of titanium bases on osteogenic differentiation of valve`s interstitial cells and to analyze the methods of express diagnostics of artificial implant materials.

Methods. The study examined titanium samples with three different coatings. The interstitial cells of the aortic valve were cultured on the studied materials. Using the PCR-RT method, the level of changes in the expression of osteogenic markers RUNX2, COL1a1, ALP under the influence of osteogenic stimuli was analyzed. To determine the level of calcification in cells, we stained the samples with alizarin red and conducted spectrophotometric analysis. Several RNA isolation methods were evaluated.

Results. The most optimal RNA isolation method involves the use of a poly(A) tails. Moreover, we developed a protocol for the absorption of calcium from composite materials. We noted multidirectional changes in the expression of RUNX2, ALP, and COL1A1 genes in cells cultured on coated titanium samples compared to controls. The intensity of expression differed depending on the type of coatings; these data correlated with the intensity of staining with alizarin. Thus, different types of coatings affect the processes of osteogenic differentiation in cells in different ways.

Conclusion. The duration of express diagnostics was 21 days and included the study of gene expression of osteodifferentiation markers at a time point of 96 hours and staining with alizarin red on day 21 from the start of osteodifferentiation. It has been established that polymer coatings affect osteogenic differentiation, however, all the studied coatings cannot be recommended for valve prostheses, it is advisable to use them to enhance osteogenic processes.

Highlights

• The number of coronary bypass surgeries in the Russian Federation in 2021 has remained below the 2019 indicator.
• The highest territorial accessibility of open-heart surgery is noted in the Volga Federal District.
• There number of open-heart surgeries performed does not depend on the capacity of the medical organization.

Aim. To study the changes in the number and outcomes of coronary artery bypass surgeries in the Russian Federation during 2019–2021 on the example of the Central, Volga and North Caucasus Federal Districts.

Methods. We used data from the following sources: official statistics published by the Ministry of Health of the Russian Federation (Statistics Form No. 14); data published by Federal State Statistics Service; data on patients from “A.N. Bakulev National Research Center for Cardiovascular Surgery”; official statistics published by the Organization for Economic Co-operation and Development; and the available literature data. Taking into account the demographic characteristics of the regions, we calculated the number of surgeries per 100.000 citizens.

Results. The total number of CABG decreased by 18.7% in 2020 (from 9 739 in 2019 tо 7918 in 2020) in the studied subjects of the Russian Federation. There was an increase in the number of direct myocardial revascularization in 2021 by 8,6% compared to 2020. However, this index was still 11.7% lower as compared to 2019. The number of CABG per 100,000 population in the regions was 24,6 ± 18,6; 21,0 ± 16,2 and 22,9 ± 17,8 in these years, respectively. At the same time, it should be noted that the decrease in the number of open-heart surgeries did not depend on the capacity of medical organization. The analysis also showed that in 2020, the average mortality rate during CABG increased by 5.6% compared to 2019 (from 1.8 ± 1.5 to 1.9 ± 1.6).

Conclusions. In 2020, the number of planned coronary artery bypass surgeries in the Central, Volga and North Caucasus Federal Districts of the Russian Federation decreased by 18,7% on average compared to 2019 with a slight positive trend in subsequent years. One of the most likely reasons is coronavirus disease pandemic.

Highlights

The article presents a review of literature data devoted to the latest changes in the epidemiology and features of transthyretin (ATTR) amyloidosis. The review highlights factors that increase the likelihood of a patient having this disease, emphasizing the group of high-risk patients, and discusses issues of non-invasive diagnostic management of a patient with cardiac amyloidosis in clinical practice.

Abstract

Transthyretin (ATTR) amyloid cardiomyopathy is a severe infiltrative heart disease that followed by heart failure (HF) with a preserved and moderately reduced left ventricular ejection fraction and a restrictive phenotype. It negatively affects the prognosis and quality of life of patients. In the absence of timely treatment, the median survival is 2–3.5 years for different forms of ATTR amyloidosis. Advances in noninvasive cardiac imaging, availability of genetic testing, identification of «high risk» groups of patients with amyloid cardiomyopathy, and the emergence of new methods of treatment have increased the detection rate of the disease, but only at its later stages (when a patient suffers from heart failure and pronounced hypertrophic cardiomyopathy), which negatively affects the prognosis of patients. Therefore, an interdisciplinary approach, vigilance, and timely diagnostics are crucial for both ATTR amyloidosis and the problem of heart failure in general.

Highlights

• The atherosclerotic process is caused by phenotypic heterogeneity and cellular plasticity of the immune system and vascular wall.
• Somatic mutations and clonal hematopoiesis with uncertain potential demonstrate a close association with cardiovascular diseases and acute vascular events.

Abstract

The last decade of cardiogenetic studies focused on inherited germline mutations. Recently researchers demonstrated a significant role of cellular heterogeneity, somatic mosaicism, and clonal hematopoiesis in the risk of coronary disease and acute vascular disorders. Up-to-date technologies, such as single-cell sequencing and mass cytometry, have made it possible to reveal fundamentally new mechanisms for the development of cardiovascular diseases. This review discloses cutting-edge data on atherosclerosis and vascular disorders, focusing on cellular heterogeneity, somatic mosaicism, and clonal hematopoiesis.

Highlights

This review precedes the planned study and is an analysis of the available works on the role of pectinate muscles in the development of morphological and functional conditions of intracavitary hemodynamics in the heart.

Abstract

This review presents an analysis of recent publications on the assessment of the functional role of the pectinate muscles in the development of morphological and functional conditions of blood flow in the atria. The results of this analysis helped us to identify a number of topics for further research. The results of this research should help us develop new diagnostic systems and criteria in cardiology field, and assist in furthering research regarding the development of anatomically and physiologically substantiated valve prostheses, taking into account the individual characteristics of the hydrodynamic patterns of blood flow.

Highlights

• The creation of a domestic bank of standardized endothelial cell cultures (arterial, venous, microvascular, valvular, and lymphatic lineages) and the development of an import-substituting technological process for working with them is a key task to enable next-generation endothelial physiology studies.
• The cultural and molecular justification for selecting the best source of autologous endothelial cells with the highest regenerative potential (in particular in context of their proliferative and angiogenic capacities) represents a primary task for endothelial physiology.
• The research methodology for studying endothelial cell physiology under normal and pathological conditions includes assessing their proliferative and angiogenic properties and molecular profiling.

Abstract

One of the primary goals in studying endothelial physiology is the development of a biomedical cell product in the form of autologous endothelial cells with high regenerative potential. These cells would be used to stimulate angiogenesis in ischemic tissues and to coat tissue-engineered constructs (such as vascular grafts) for their endothelialization prior to implantation into the human body. This review examines the key applied aspects of endothelial physiology aimed at achieving these objectives, including the selection of endothelial cell sources for modeling various pathological processes and regenerative medicine, the creation of a bank of standardized endothelial cell lines with various differentiation directions to enhance the effectiveness of this modeling, methods to ensure adhesion, sustained proliferation, and physiological functioning of primary endothelial cells in culture, as well as the methodology for assessing proliferative and angiogenic activity in the context of analyzing endothelial cell resistance to external influences. The discussion emphasizes the need for a comparative analysis of the regenerative properties of microvascular endothelial cells from subcutaneous adipose tissue, colony-forming endothelial cells from peripheral venous blood, and endothelial cells differentiated from induced pluripotent stem cells. Additionally, the current state of import substitution for various components of endothelial cell technology is reviewed, including standardized cell lines, functional protein coatings, and three-dimensional matrices for angiogenesis assessment, culture media for isolation, cultivation, and experimentation with endothelial cells, as well as other reagents for the isolation and subculturing of endothelial cells, culture dishes, and antibodies for flow cytometric and fluorescent microscopic immunophenotyping. The review also analyzes the prospects for import substitution of missing components, such as certain endothelial cell lines, magnetic beads with immobilized antibodies for their isolation, and antibodies conjugated with various fluorophores.

Highlights

Adherence to CPAP therapy in patients with severe obstructive sleep apnea and cardiovascular diseases can be predicted using a model that includes the desaturation index and anxiety.

Aim. To evaluate the results of CPAP therapy (Continuous Positive Airway Pressure) in cardiac patients with severe obstructive sleep apnea (OSA) and to identify factors predicting adherence to CPAP therapy using prospective follow-up data.

Methods. The study included 52 patients with severe OSA (82.7% men, 55.6 ± 8.8 years) examined in 2019–2022, the data was collected from the registry of cardiac patients with OSA (n = 138). Arterial hypertension was diagnosed in 96.1% of patients, coronary artery disease (CAD) was diagnosed in 42.3% of patients, chronic heart failure was diagnosed in 86.5% of patients. Clinical and laboratory parameters, data from OSA screening scales, overnight pulse oximetry, diagnostic scales of anxiety, depression, quality of life and cognitive function were assessed before the start of CPAP therapy and 12.3 ± 7.0 months after its start. 8 patients adequately used CPAP therapy (group with CPAP) and 44 patients either refused to use it or used it inadequately (group without CPAP).

Results. When comparing the groups at baseline, there were no differences in age, gender, body mass index and clinical characteristics. Predictors of adherence to CPAP therapy were a higher desaturation index (odds ratio (OR) 1.264, 95% confidence interval (CI) 1.017–1.569, p = 0.034) and manifestations of anxiety (OR 1.056, 95% CI 1.01–1.115, p = 0.049). The sensitivity of the model was 85.7%, specificity was 78%, and area under the ROC curve was 0.843 (95% CI 0.712–0.974, p = 0.004). There were no new cases of CAD in the group with CPAP, in the group without CPAP, 5 new cases of CAD developed during follow-up.

Conclusion. Adherence to CPAP therapy in patients with severe OSA and cardiovascular disease can be predicted with high sensitivity and specificity using the model that includes the desaturation index and anxiety. Supposedly, CPAP therapy is relevant for secondary prevention of coronary artery disease.

Highlights

Patients with pulmonary embolism in the Kemerovo region have severe comorbidities and are older than those included in the Russian registry, however, these patients show a significantly lower rate of thromboembolism provoked by surgical interventions. The management of this group of patients in Kuzbass region includes active recanalization of the pulmonary artery branches with the help of fibrinolysis and surgical methods of thrombus removal.

Aim. To comparatively analyze the features of diagnosis, treatment and outcomes in patients with pulmonary embolism (PE) from the Kuzbass patient registry and the results of the “SIRENA” Russian registry.

Methods. The registry-based prospective single-center study included 185 patients with PE residing in the Kemerovo region. Moreover, we analyzed the data of 609 patients from the SIRENA registry (observational prospective multicenter study) who underwent a standard examination for patients with PE. Statistical processing of the study results was carried out using the MedCalc Version 16.2.1 software package (Softwa, Belgium).

Results. Patients from the Kemerovo region have more adverse risk factors for pulmonary embolism compared with patients from the SIRENA registry: chronic heart failure (CHF) (23.30% versus 50.80%), coronary artery disease (CAD) (8.50%) versus 36.20%), acute stroke) in history (9.90% versus 12.40%), arterial hypertension (AH) (65.70% versus 78.40%), type 2 diabetes mellitus (type 2 diabetes) (15.20% vs. 20.50%), chronic kidney disease (CKD) (9.00% vs. 12.40%).

Conclusion. Patients with PE in the Kemerovo region are characterized by a higher prevalence of cardiovascular diseases (coronary artery disease, arterial hypertension, chronic heart failure, venous thromboembolism in the anamnesis), older age, predominance of women, more frequent use of fibrinolysis compared with patients included in the SIRENA Russian registry. Only in Kuzbass specialists used surgical methods of thrombus removal.

Highlights

• Extracellular vesicles (EVs) are membrane objects with inner and surface molecular cargo performing various biological functions associated with intercellular connection. Therefore, they are a promising object for diagnostic and therapeutic purposes.
• THP-1 is a human leukemia monocytic cell line and a classic model for monocyte/macrophage function and differentiation. Spectrum of THP-1-derived extracellular vesicles are heterogenic and understudied.
• The review presents the comparison of EVs produced by stimulated THP-1 cells.

Aim. To quantitatively and qualitatively characterize THP-1-derived extracellular vesicles in depending on type and dose of used stimulators.

Methods. THP-1 cells were stimulated by PMA (4-phorbol 12-myristate 13-acetate), LPS (lipopolysaccharide) and TNF (tumor necrosis factor). Cell activation and viability were assessed by the expression of CD54 and DAPI (4′,6-diamidino-2-phenylindole) staining respectively. Size and quantitative characteristics were assessed using nanoparticle tracking analysis (NTA). Qualitative assessment of surface molecular markers (CD9, CD63, CD54) was performed by high-sensitivity multicolor flow cytometry (hs-FCM).

Results. TNF and PMA stimulation in medium and high doses led to an increase in quantity of small EVs (30–150 nm) that express tetraspanins CD9 and CD63. Low doses of TNF and PMA did not lead to a significant increase of EVs compared to control. There was a decrease in amount of small EVs (30–150 nm) in LPS-stimulated probes. However, medium and high doses of LPS stimulated selective CD63-positive EVs secretion with absence of CD9-positive events. Cell activation assessed by CD54 expression have been dose-dependent and was most prominent under PMA stimulation.

Conclusion. Every type of THP-1 cell activators promotes changes in the functional activity of cells towards production of extracellular vesicles. Moreover, we have demonstrated quantitative and qualitative difference in THP-1-derived vesicles under different stimulation conditions. Medium and high doses of PMA and TNF were the most relevant in stimulation of vesiculation. LPS-stimulation could be used to selectively obtain CD63-positive fraction of EV.

Highlights

• To achieve a 10% (i.e., 1.1-fold) increase in ionized calcium level in the serum-free cell culture medium and Wistar rat serum, 10 µg/mL calcium (e.g. with CaCl₂) should be added; this corresponds to the upper quartile of ionized calcium in the human population.
• Incubation with ionized calcium or calciprotein monomers (10 µg/mL calcium) does not induce any pathological effects in primary arterial endothelial cells, although calciprotein monomers are internalised by endothelial cells similar to calciprotein particles.
• Incubation of primary human coronary artery endothelial cells with calciprotein particles (10 µg/mL calcium) triggers an increased expression of VCAM1, ICAM1, and SELE genes (i.e., those encoding cell adhesion molecules) as well as IL6, CXCL8, and CXCL1 genes (i.e., those encoding pro-inflammatory cytokines), together indicating endothelial activation.

Aim. To compare the pathological effects of ionized calcium, calciprotein monomers (CPMs), and calciprotein particles (CPPs) after their addition to endothelial cells.

Methods. CPMs and CPPs were synthesized by the supersaturation of albumin-supplemented NaCl solution with calcium (by adding CaCl₂) and phosphate ions (by adding Na₂HPO₄). CPMs and CPPs were separated by sequential ultracentrifugation (to isolate CPPs) and ultrafiltration (to separate CPMs and free mineral ions). Calcium concentration in CPMs and CPPs was measured by an o-cresolphthalein complexone-based colorimetric assay. Internalization of fluorescent-labeled CPMs and CPPs by endothelial cells was interrogated by confocal microscopy after their 1-hour co-incubation under flow. Primary coronary artery and internal thoracic artery endothelial cells were incubated with ionized calcium (CaCl₂), CPMs, or CPPs (10 µg/mL calcium) upon reaching confluence (n = 6 wells per group) to simulate a 10% increase in ionized calcium concentration. After 24 hours, we collected RNA and analyzed the gene expression pattern by reverse transcription-quantitative polymerase chain reaction.

Results. CPMs and CPPs were internalized by endothelial cells as soon as after 1 hour of co-incubation under flow. Treatment with ionized calcium or CPMs (10 µg/mL calcium) did not cause any pathological effects in arterial endothelial cells. In contrast, CPPs promoted an expression of VCAM1, ICAM1, and SELE genes (i.e., those encoding cell adhesion molecules) as well as IL6, CXCL8, and CXCL1 genes (i.e., those encoding pro-inflammatory cytokines) in primary human coronary artery endothelial cells, together indicating their pro-inflammatory activation.

Conclusion. Pathological effects of calcium stress are defined by the calcium source and not simply by its concentration.

Highlights

• Hybrid decellularization of allografts in scCO₂ with a small addition of the detergent Tween 80 is very effective in terms of reducing calcification.
• A special coating based on chitosan nanoparticles obtained and applied from the water/subcritical CO₂ system makes it possible to significantly reduce calcification of the standard GA-stabilized pericardium.

Aim. To conduct a comparative analysis of anticalcification properties of implants based on allo- and xeno-tissue after treatment in super- (scCO₂) and subcritical carbon dioxide.

Methods. We have performed a modification of the following structures: a) rat vessels by hybrid decellularization in scCO₂ with detergents b) calf pericardium by direct coating of chitosan nanoparticles with silver from the water/carbon dioxide system under pressure (carbonic acid). Following that we have performed an in vivo calcification study on rats (n = 10 for each type of sample). To model the behavior of biomaterial in the human body, we have implanted allogeneic modified aorta into rats. To determine the calcium in the extracted samples, we have used spectrophotometric method to estimate the concentration of calcium. To assess the cytotoxicity of nanocoated pericardium samples, we have measured the impedance of cells populated on biomaterial samples was used.

Results. The treatment of rat vessels in supercritical CO₂ reduces the deposition of calcium salts by 30% compared to baseline allografts. Pericardium coated with chitosan nanoparticles can be characterized by a significant decrease in calcification – the biomatrix coated with Cht_Ag/TPP contains only (0.015 ± 0.005) wt.% of calcium salts, which is significantly lower compared with GA-stabilized pericardium with (0.070 ± 0.015) wt.% after the experiment. Biocomposites with nanocoating are characterized by similar cellular index compared to a sample of uncoated xeno-tissue. All samples exhibit a cytostatic effect, including nanocoated samples (Cht_Ag/TPP), which proves the encapsulation of silver in the polymer matrix and ensures the absence of negative effects on the body during normal functioning. The coating of biomatrix based on hollow chitosan nanoparticles contributes to an increase in cell proliferation.

Conclusion. The use of supercritical technology for the purification of biomaterials, as well as the creation and application of polymer protective coatings in order to enhance the ability to suppress calcification looks very promising. At the same time, the use of such a “green” approach to the modification of bioprostheses is a more advantageous strategy relative to the standard generally accepted methods of their chemical modification.

Highlights

The original method of forming a hydrogel anti-trombogenic drug coating on the inner surface of a small diameter vascular graft did not affect the baseline structural, physical and mechanical properties of the product. Loading iloprost and heparin into the coating significantly improved the hemocompatibility of the material with respect to inhibition of platelet aggregation.

Aim. To evaluate structural, physical and mechanical characteristics of a drug-eluting tissue-engineered vascular graft and the functional activity of its coating.  

Methods. Vascular grafts based on polycaprolactone and polyurethane with growth factors were manufactured by emulsion electrospinning. The modified coating on the inner surface of the graft consisted of a hydrogel layer loaded with iloprost and heparin. The surface structure was studied by scanning electron microscopy, physical and mechanical characteristics were studied using a universal Zwick/Roell testing machine, surface properties were studied by contact angle measurements and wettability. The functional activity of the modified coating was assessed with the help of indicators of maximum platelet aggregation upon contact with the material.

Results. The results showed that forming the coating did not change the architectonics of the polymer matrix surface. The structure of the graft was represented by randomly directed micro- and nanofibers with heterogeneous pores and without deformation of material; the surface of the graft retained hydrophilicity (θ = 68.6 ± 11.85). We confirmed the success of surface modification of vascular graft by observing extremely low values of the maximum platelet aggregation upon contact of platelet-rich plasma with the surface of the graft (8,5 (7,2; 12,2) %). Depending on the conditions of their incubation in saline solution, we have noted the prolongation of the drug-releasing activity.

Conclusion. The formation of anti-thrombogenic coating on the surface of the PCL/PU/GFmix/^Ilo/Hep vascular graft did not negatively affect the structural characteristics or physical and mechanical properties, moreover, it significantly improved hemocompatibility in terms of inhibition of platelet aggregation. 

Highlights

The review is dedicated to analysis of available data on diagnostic significance of extracellular vesicles and their nucleic acids and proteins cargo. The analysis is focused on clinical application of testing systems in various pathological conditions and illnesses. In particular, diagnostic value of miRNA levels in different types of cancer are also presented.

Abstract

The increased number of performed studies investigating biological nature of extracellular vesicles, as well as their participation in many biological and pathological processes, determines the need to translate the acquired knowledge into practical medicine. The development of diagnostic and prognostic algorithms based on the study of the level of target extracellular vesicles in patients with various pathological conditions is the most promising way of translation of this knowledge. This review attempts to systematize the available data on the experience and/or prospects for the use of certain classes of extracellular vesicles in the diagnosis of the most common diseases and pathological conditions (ischemic and reperfusion injuries of tissues and organs, infectious and non-infectious inflammatory diseases, malignant tumors, etc.). Potentially applicable extracellular vesicles for diagnostic purposes are considered separately in terms of their protein composition, as well as in terms of the repertoire of transferred nucleic acids (primarily miRNAs).