Cellular cardiomyoplasty for refractory angina: experimental rationale.

Cellular cardiomyoplasty for refractory angina: experimental rationale. | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Cellular cardiomyoplasty for refractory angina: experimental rationale. Svetlana Gramatiuk, Yulia Ivanova, Karine Sargsyan, Igor Kryvoruchko, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4024504/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This experimental study aimed to investigate the therapeutic potential of cellular cardiomyoplasty in the context of refractory angina using a rodent model. The study comprised five groups of animals subjected to different interventions, including "empty" injections, autologous mesenchymal stem cell (MSC) injections via various routes, and untreated controls. Stress load modeling was employed to assess the hemodynamic response, mainly focusing on heart rate changes during isopropyl norepinephrine loading. Results revealed a positive chronotropic effect in all groups, with varying severity. Intact rats exhibited a significant increase in heart rate, while animals in the "empty" injection group demonstrated a less pronounced response. Interestingly, those treated with MSCs, either through direct myocardial injection or intravenous and intracavitary routes, exhibited notable variations in stress-induced heart rate changes. By the third minute of the experiment, a decrease in heart rate was observed across all groups, stabilizing at 490–495 beats/min. Notably, the groups with untreated myocardial infarction and "empty" injections displayed an inability to compensate for required blood flow during stress, indicative of potential challenges in these conditions. These findings suggest that cellular cardiomyoplasty, particularly with autologous MSCs injected into the myocardium, may influence the stress-induced changes in heart rate. However, a comprehensive evaluation of additional cardiovascular parameters and further exploration of therapeutic outcomes are warranted to elucidate the potential clinical relevance of these observations. This study contributes valuable insights into the experimental rationale and hemodynamic effects of cellular cardiomyoplasty, laying the groundwork for future investigations in refractory angina and myocardial infarction. cellular cardiomyoplasty autologous mesenchymal stem cell cardiovascular parameters Introduction Ischemic heart disease (IHD) is the most common cause of cardiovascular disease in terms of complication frequency and number of fatal outcomes. In the United States, it is the cause of every fifth death [ 1 ]. In Ukraine, the diagnosis of IHD is established for approximately 400,000 patients annually [ 2 , 3 ]. Traditional methods of treating this category of patients, which currently exist, are: The available treatments for ischemic heart disease include medication, direct myocardial revascularization (coronary artery bypass grafting or angioplasty with stenting), and heart transplantation. However, current medication therapy is often insufficient to prevent myocardial remodeling [ 4 , 5 ]. Heart failure (HF) caused by IHD or cardiomyopathies is a severe disease with a poor prognosis. Despite the wide range of pharmaceuticals and surgical interventions available, there remains a significant number of patients with angina for whom surgery is not possible for various reasons and for whom drug therapy is insufficiently effective [ 6 , 7 ]. The definition of refractory angina (RA) was first proposed in 2002 by a joint group of the European Society of Cardiology for treating refractory angina. It is a chronic condition (lasting more than three months) characterized by the presence of angina caused by insufficient coronary blood flow (due to coronary artery disease), accompanied by severe clinical symptoms. Recent studies in stem cell biology have fundamentally changed all concepts of myocardial regenerative capacity and have become the beginning of a new therapeutic direction - cellular cardiomyoplasty. Patients who cannot be controlled with combined drug therapy at the maximum tolerated doses and are unable to undergo myocardial revascularization (percutaneous coronary angioplasty or coronary artery bypass grafting) [ 8 , 9 ]. The text discusses stem cell transplantation as a potential therapy for patients with chronic ischemic heart disease. However, no large-scale studies have evaluated the effectiveness of stem cell implantation in patients with heart failure. Many fundamental issues in cell therapy remain unresolved, including homing mechanisms, differentiation and engraftment of transplanted stem cells, the role of cell fusion, and the mechanisms by which transplanted cells affect the function and metabolism of the heart muscle. The most effective method of delivering cells to the myocardium, the timing of cardiomyoplasty, the number of cells in the transplant, and methods of preparation are also topics of discussion. However, much research has been devoted to these issues [ 10 , 16 ]. This experimental study aimed to investigate the therapeutic potential of cellular cardiomyoplasty in the context of refractory angina using a rodent model. The study within the framework of the academic research work at the basis of the study and the use of data were consented to by the Ethics Committee of Kharkiv National Medical University, Ukraine (Protocol No. 6, November 11, 2022) and Ukraine Association of Biobank (Protocol dd. 11/11/2021) and conducted by the Law of Ukraine "On Protection of Animals from Cruelty" and the provisions of the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes by ARRIVE guidelines. Materials and methods The experimental part of the work was carried out on 122 rats of the inbred Wistar-Cayote line weighing 200–220 g, kept in a vivarium in compliance with ARRIVE recommendations. We used the Wistar-Kayota breed because it is an inbred breed that minimizes the rejection reaction, given its genetic homogeneity. The animals were used in the experiment by the rules regulated by the European Convention for the Care and Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes (Strasbourg, 1986), the Council of the European Community Directive of 24.11.86 and the order of the Ministry of Health of Ukraine No. 32 of 22.02.88. Surgical interventions were performed in an experimental operating room under ketamine anesthesia (12.5 mg/100 g of body weight intramuscularly). MI induction was performed by stitching and ligation of the anterior interventricular artery. Only animals with transmural MI, which ECG and ultrasound confirmed, were included in the further experiment. The effect of cellular cardiomyoplasty on the course of MI was studied in 100 surviving animals (22 animals died in the first hours after modeling the pathological condition due to the development of life-threatening arrhythmias). The animals included in the study were withdrawn from the experiment by decapitation (under general anesthesia). The animals were divided into five groups (20 animals in each group): in group 1, no treatment was performed; in group 2, "empty" injections were performed into the myocardium in the area of the ischemia zone, which was determined macroscopically, in group 3, autologous mesenchymal stem cells (MSCs) were injected at a dose of 10 million cells. In group 4, MSCs were injected intravenously at the same dose by puncture of the tail vein; in group 5, MSCs were injected into the left ventricular cavity (LVC) by puncture and catheter through the right femoral artery (this was done to create the maximum concentration of MSCs in the coronary vessel mouth and to simulate intracoronary injection. MSCs were obtained from the peripheral blood of animals according to the following procedure. A syringe containing 0.5 ml of phosphate-buffered saline, 50 units/ml of heparin, and 0.25 mg/l of gentamicin sulfate was used to collect 0.5 ml of blood from the tail vein of the animal. The volume of blood taken from the animals was acceptable and safe, as it did not exceed 10% of the circulating blood volume (13.75–17.50 ml for rats, based on the recommendations for safe blood collection in mice and rats of the US National Institutes of Health). The cell suspension was centrifuged at 1500 rpm for 5 min; the cell pellet was resuspended in erythrocyte lysis solution (114 mM ammonium chloride, 7.5 mM bicarbonate, 100 µM EDTA) for 3 min and centrifuged again. The haemolysed supernatant was removed, and the cell pellet was resuspended in DMEM containing 10% fetal calf serum (HyClonegold, USA), insulin 0.4 µM, and 0.25 mg/l gentamicin sulfate. The resulting cells were seeded in culture vials and transferred to a CO 2 incubator with a 5% carbon dioxide concentration and 95% humidity. 2 days after the primary culture was isolated, the non-attached cell suspension was removed, and the remaining cells with fibroblast-like morphology were continued to be cultured. The culture medium was replaced with fresh medium every 3 to 4 days. After 75–80% monolayer formation, the cells were washed once with Versailles solution, removed with Versailles solution with 0.25% trypsin solution, resuspended in growth medium, and poured into new culture dishes. The cellular material, flattened fibroblast-like cells (mesenchymal stromal cells) fixed on plastic, retained population activity, and did not contain dead cells, was considered suitable for use. The stromal origin of the stem cells was confirmed by immunohistochemical method in culture by detecting type I collagen using rabbit monoclonal antibodies. MSCs of the first and second passages were used for the experiment. The viability of MSCs was determined before their injection by trypan blue staining. For this purpose, a 0.1% trypan blue solution was used for the cell suspension, and after 2–3 min, live cells were counted in a phase-contrast microscope without staining their membranes. Markers of neoangiogenesis in the blood serum were studied in laboratory animals in the dynamics. The nitric oxide (NO) content in the blood plasma was judged by the number of stable end metabolites of NO, namely NO 2 ¯ + NO 3 ¯ (UNOX). The concentration of vascular endothelial growth factor (VEGF) was measured using a Luminex dual-laser flow cytometric fluorescence analyzer (Luminex Corporation, USA) based on Simplex ProcartaPlexTM reagent kits (Affymetrix, USA). The level of endothelin-1 in blood plasma was determined by enzyme-linked immunosorbent assay using the Endotelin (1–21) kit from Biomedica (Austria) on a Stat Fax 2100 enzyme-linked immunosorbent assay. Study period: 1, 6, and 24 hours after induction of the pathological condition and treatment, as well as on days 7 and 30 of the study. Echocardiography was performed using a GE Vivid newborn ultrasound machine (USA) with a 12 MHz transducer. Study period: 1 and 3 months of the experiment. The following rat heart parameters were studied in different experimental groups: LVIDd - end-diastolic left ventricular internal dimensions; LVIDs - end-systolic left ventricular internal dimensions; FS - fractional shortening; EF - ejection fraction; SV - stroke volume. Electrocardiography (ECG) was performed on an IVF 1T apparatus on day 30 of the experiment. Used the standard statistical software package SPSS 20.0 for Windows for statistical data processing. Statistical data processing was performed using a non-parametric Kruskal-Wallis test and Statistica software (StatSoft, Inc., USA). Considered differences statistically significant at p < 0.05. Results of the experimental block of studies When studying the levels of vasoconstrictor endothelin-1, vasodilator nitric oxide, and endothelial vascular growth factor in the dynamics one month after modeling MI, determined a general pattern for all three indicators: they reached their maximum value one day after modeling MI (Table 1 ). Table 1 Changes in the concentration of angiogenesis markers in rat blood serum Indicators Control The frame of the study Animal groups 1 hour 6 hours 24 hours Seven days 30 days NO (mg/ml) (n = 20) 0.58 ± 0.03 0.86 ± 0.04 *** 0.92 ± 0.03 1.12 ± 0.05 *** 0.99 ± 0.04* 0.88 ± 0.03 * 1 0.89 ± 0.05 *** 1.04 ± 0.04 * 1.26 ± 0.03 *** 1.24 ± 0.04 0.96 ± 0.05 *** 2 VEGF (pg/ml) ( n = 20) 66.98 ± 12.47 70.21± 11.36 126.72± 24.05* 220.45± 22.13* 134.89± 25.24 89.74± 21.38 1 71.42± 13.45 134.86± 28.11* 288.22± 23.46*** 189.57± 28.47* 132,74± 19,87 2 Endothelin-1 (fmol/ml) (n = 20) 5.1 ± 0.4 10.6 ± 0.7 *** 12.8 ± 0.5 * 12.9 ± 0.4 8.8 ± 0.3 *** 5.3 ± 0.4 *** 1 10.4 ± 0,5 *** 12.8 ± 0.6 ** 9.1 ± 0.3 *** 6.9 ± 0.4 *** 5.1 ± 0.2 *** 2 Note: when compared with the previous indicator: * - p < 0.05; ** - p < 0.01; *** - p < 0.001; 1 - untreated MI group; 2 - MI + i.v. MSCs transplantation. The experimental data obtained indicated a decrease in myocardial ischemia after MSCs transplantation, which led to a decrease in the level of such a potent vasoconstrictor as endothelin1. When studying the dynamics of VEGF concentration, which reflects the intensity of angiogenesis, it was found that up to 6 hours after MI induction, this indicator did not differ from control values. After 6 hours in the group of animals with MI, the VEGF content increased and reached its maximum peak by the end of the 1st day. By the end of the experiment, the level of VEGF decreased to (89.74 ± 21.38) pg/ml, which did not differ from normal values (t = 0.91; p > 0.05). In the group of animals with MI + MSCs transplantation, a significant increase in VEGF levels was also observed by 6 hours of the study, reaching its maximum by the end of the 1st day; its level decreased slightly and by the end of the experiment was (132.74 ± 19.87) pg/ml, which was two times higher than normal values (t = 2.8; p < 0.05). When analysing the effect of MSCs transplantation on vascular tone parameters after MI, it was found that cellular cardiomyoplasty had a pronounced vasodilator effect from the first day. This was reflected in an increase in NO levels with a simultaneous decrease in the content of the vasoconstrictor endothelin 1. It is characteristic that the increased nitric oxide content persisted until the experiment's end. When studying the concentrations of nitric oxide derivatives in groups of animals with different methods of MSCs administration, it found that after one month in rats of the 1st and 2nd groups, their increase to (0.88 ± 0.03) and (0.85 ± 0.02) µg/ml, respectively, occurred (t = 6.4, p < 0.05). In the 3rd, 4th, and 5th groups, a more significant increase in the studied indicator was found: (0.99 ± 0.04), (0.96 ± 0.05) and (0.92 ± 0.03) µg / ml, respectively, while in the 3rd group, the NO content was higher than in the 5th group (t = 1.99, p < 0.05). Table 2 Concentrations of angiogenesis markers in rat blood serum 30 days after MI modeling Indicators Control Animal groups 1st (n = 20) 2nd (n = 18) 3rd (n = 20) 4th (n = 20) 5th (n = 20) NO (mg/ml) 0.58 ± 0.03 0.88 ± 0.03 *** 0.85± 0.02*** 0.99 ± 0.04*** 0.96 ± 0.05 *** 0.92 ± 0.03 *** VEGF (pg/ml) 66.98 ± 12.47 0.88 ± 0.03 *** 0.85± 0.02*** 0.99 ± 0.04*** 0.96 ± 0.05 *** 0.92 ± 0.03 *** Endothelin-1 (fmol/ml) 5.1 ± 0.4 5.9 ± 0.2* 5.8 ± 0.2* 4.9 ± 0.2 5.1 ± 0.2 5.0 ± 0.3 Note: in comparison with the control: * - p < 0.05; ** - p < 0.01; *** - p < 0.001. A similar trend was observed concerning the concentration of VEGF: in groups 1st and 2nd, its level did not exceed average values; in groups 3rd, 4th and 5th, the studied indicator was significantly higher than usual and amounted to (144.22 ± 14.59), (132.74 ± 19.87) and (111.43 ± 12.22) pg/ml, respectively, while, by analogy with the level of nitric oxide, determined the maximum level in group 3 compared to animals in group 5 (t = 1.97, p < 0.05). Analysis of the concentration of endothelin 1 showed that in the 3rd, 4th, and 5th groups one month after the experiment's induction, the level of the studied indicator did not differ from the norm. In the 1st and 2nd groups, it was slightly higher than normal values and amounted to (5.9 ± 0.2) and (5.8 ± 0.2) fmol/ml, respectively. When studying the final diastolic LV internal diameter, it was found that in the 1st group of animals, it increased from (6.16 ± 0.12) to (7.19 ± 0.16) mm, t = 5.15, p < 0.001. A similar trend was determined in the second group: this indicator increased to (7.21 ± 0.14) mm, t = 5.7, p < 0.001. In animals of groups 3 and 4, LVIDd remained unchanged at t = 0.12 and 0.98, respectively. In group 5, LVIDd also increased to (6.81 ± 0.15) mm at t = 3.4, p < 0.01. In the study of LVIDs, it was found that in the 1st group of rats, it increased from (2.82 ± 0.18) to (3.82 ± 0.11) mm at t = 4.7, p < 0.05. In animals of the 2nd group, this indicator increased to (3.76 ± 0.13) mm at t = 4.2, p < 0.01. It should be noted that in the 3rd and 4th groups, LVIDs remained unchanged, and in the 5th group, it increased to (3.62 ± 0.13) mm at t = 3.5, p < 0.05. At the same time, LVIDs in group 5th did not differ significantly from those recorded in rats of groups 1st and 2nd and were higher than in group 3rd at t = 4.0, p < 0.01. Thus, the best values of LVIDd and LVIDs were obtained in animals of groups 3rd and 4th (Table 3 ). Table 3 Ultrasound parameters of heart function in rats Indicators Animal groups Control 1st (n = 20) 2nd (n = 20) 3rd (n = 20) 4th (n = 20) 5th (n = 20) LVIDd, mm 6.16 ± 0.12 7.19 ± 0.16 7.21 ± 0.14 6.18 ± 0.11 6.34 ± 0.14 6.81 ± 0.15 LVIDs, mm 2.82 ± 0.18 3.82 ± 0.11 3.76 ± 0.13 2.85 ± 0.14 2.93 ± 0.15 3.62 ± 0.13 SF, % 45.3 ± 1.7 26.8 ± 0.8 27.3 ± 0.7 45.0 ± 0.9 42.6 ± 0.12 31.2 ± 0.9 LVEF, % 76.9 ± 2.5 55.3 ± 3.4 56.2 ± 3.2 75.8 ± 3.2 70.6 ± 2.3 66.5 ± 3.3 SV, ml 0.25 ± 0.08 0.13 ± 0.03 0.15 ± 0.05 0.24 ± 0.07 0.21 ± 0.02 0.18 ± 0.02 Note: The statistical significance of the figures is given in the text. When comparing the shortening fraction (SF), it was found that in animals of the 1st and 2nd groups, this indicator decreased by (26.8 ± 0.8) and (27.3 ± 0.7)%, respectively, at t = 9.8, p 0.05. In group 4th, the SF was slightly lower than normal and was equal to (42.6 ± 0.12)% at t = 1.98, p < 0.05. In the 5th group of animals, the SF was equal to (31.2 ± 0.9)%, below typical values at t = 7.3, p < 0.001. At the same time, the studied index was higher in group 5th compared to the results obtained in animals of groups 1st and 2nd at t = 3.7 and 3.4, respectively, p < 0.05, and higher than in groups 3rd and 4th at t = 10.8 and 12.6, respectively, p < 0.001. In addition, in animals of group 3rd, SF was higher than in group 4 at t = 2.6, p < 0.05. A similar trend was observed in the analysis of changes in left ventricular ejection fraction (LVEF) dynamics. Thus, in group 1st, it decreased from (76.9 ± 2.5) to (55.3 ± 3.4)% at t = 5.1, p < 0.05. In the 2nd group, the same picture was observed, with no difference in the indicators between groups 1st and 2nd. In animals of group 3rd, LVEF did not differ from the norm, and in group 4th, this indicator was below the norm and was equal to (70.6 ± 2.3)% at t = 1.9, p < 0.05. In animals of group 5th, LVEF also decreased to (66.5 ± 3.3) at t = 2.5, p < 0.05. It should be noted that the LVEF in group 5th was higher than in groups 1st and 2nd at t = 2.4 and 2.2, p < 0.05, but lower than in group 3rd at t = 2.02, p < 0.05. The LVEF between the 4th and 5th groups did not differ significantly. The dynamics of changes in stroke volume in the experimental groups were as follows: in group 1st, SV decreased from (0.25 ± 0.08) to (0.13 ± 0.03) ml at t = 1.9, p < 0.05, and in the other groups this indicator remained unchanged. However, in group 3rd, the SV was closest to normal and was higher than in groups 1st, 2nd, and 5th of laboratory animals. Thus, the values obtained in groups 3rd, 4th, and 5th animals were the closest to normal, while in group 3rd, they practically did not differ from the norm. When modeling stress load in animals of all groups, a positive chronotropic effect was observed, with the severity of the latter differing significantly in animals of different groups. Thus, in intact rats, the absolute increase in the first minute of isopropyl norepinephrine loading was about 32 beats/min. Against the background of a high baseline heart rate (HR) in animals of the second group, the stress-induced increase was 11 beats/min. In the animals of the third group, the same indicator was 22 beats/min. By the third minute of the experiment, a decrease in HR was observed in all groups of animals with stabilization at 490–495 beats/min (Table 4 ). Table 4 Indicators of the chronotropic function of the corpus callosum under stress modeling of loading Animal groups Output HR Max HR Increase in HR Final HR Control 485 ± 43 517 ± 35 + 32 490 ± 23 1st (n = 10) 503 ± 23 * 525 ± 15 * + 22 * 495 ± 12 2nd (n = 10) 507 ± 18 * 522 ± 11 ** + 24 ** 510 ± 13 ** 3rd (n = 10) 489 ± 11 * 526 ± 8 ** + 38 * * 494 ± 10 ** 4th (n = 10) 482 ± 14 528 ± 9 ** + 39 * * 499 ± 15 ** 5th (n = 10) 501 ± 16 ** 527 ± 7 ** + 28 ** 503 ± 11 ** Note: * - significance of differences between the studied indicator and the previous data (p < 0.05); ** - significance of differences between the studied indicator and the control (p < 0.05). The qualitative response of the heart to the stress-simulating load in different groups of animals was the same; however, at a rate of + 32 beats/min, the maximum increase was closest to the norm in groups 3rd, 4th, and 5th, i.e., by + 38, +39 and + 28 bears/min, respectively. The increase in groups 1st and 2nd was only + 22 and + 24 beats/min. Thus, in the groups of rats with untreated MI and "empty" myocardial injections, compensation of the required blood flow is impossible, despite the stress, which is what the clinic of heart failure is directed to. Discussion In the current literature, the main question remains open: the mechanism of the protective effect of stem/progenitor cells. Multipotent mesenchymal stromal cells are one of the most attractive cell types for cell therapy due to their proven cardioprotective properties and low immunogenicity [ 11 , 12 ]. To date, stem/progenitor cell researchers have been divided into two groups: those who believe that after being introduced into the body, stem cells and progenitor cells differentiate and replace dead or damaged cells and those who think that the paracrine activity of stem cells and progenitor cells, i.e., the synthesis and secretion of specific signaling molecules, is the key to the therapeutic effect of these cells [ 13 , 14 ]. Both groups of scientists recognize the positive therapeutic impact of introducing stem or progenitor cells in treating pathological conditions of various organs. The study of protective effects in several studies conducted in rats has shown that introducing MSCs after experimental infarction leads to decreased volume and improved myocardium functional recovery [ 15 ]. It is recognized that MSCs can promote tissue regeneration by secreting growth factors and other cytokines (including BDNF, NGF, VEGF, and others) [ 16 ]. In addition, they can regulate the inflammatory response, which plays a vital role in postischemic myocardial damage, as SCs secrete several immunomodulatory factors, such as prostaglandin E2, TNF-a, TGF-b, IL6, and others [ 17 ]. However, the mechanisms underlying the regenerative potential of MSCs still need to be fully understood and require further study. One of the ways to study the mechanisms of SCs' influence on organ function is to directly examine the interaction between the body's cells and transplanted cells to search for chemical compounds that stem cells can secrete into the intercellular space after they are co-cultured with differentiated cells. The same applies to recipient cells, as they can respond to the presence of transplanted cells by synthesizing and secreting various biological compounds. Such "communication" between neighboring cells or distant organs within the same organism is very likely to occur. A relatively large amount of data has been obtained confirming the existence of this phenomenon [ 18 , 19 ]. However, intercellular communication as a result of the secretion of biologically active compounds and their "trapping" by neighboring cells is limited by diffusion. This limitation can be minimized in the case of the directed transfer of cellular components from cell to cell as a result of the formation of intercellular contacts. Examples of such contacts include the gap junction formed by the connexin 43 protein, which provides intercellular communication, electrical coupling of neighboring cells, and transmission of depolarisation signals [ 20 ]. Among the intercellular interactions, the migration of cytoplasmic components between contacting cells along cell processes, particularly tunneling nanotubes, deserves special attention [ 18 , 21 ]. For several differentiated cells in direct contact with progenitor cells, the exchange of cytosolic components [ 22 ] and intercellular transport of mitochondria [ 23 ] is shown. Direct intercellular contact provides the least costly way of exchanging chemical/biological information between cells; however, the efficiency of such exchange is limited by the meager ratio of stem cells to differentiated cells and, as a result, a low number of intercellular contacts. A more costly method of information exchange for cells is the secretion of biologically active molecules [ 24 ] into the intercellular space (into the culture medium in vitro and into the extracellular space/interstitial fluid in vivo). At a higher level, communication between cells in different organs can occur. In this case, certain factors are secreted into the bloodstream, which then enters distant organs via the bloodstream. Thus, cellular cardiomyoplasty improves cell metabolism, confirming the neoangiogenesis theory and the paracrine effect of cell transplantation. Therefore, MSC transplantation can optimize cardiomyocyte oxygen metabolism and stabilize aerobic and anaerobic glycolysis. Mitochondrial transport, rather than engraftment of allogeneic MSCs into tissues, can partially explain the positive therapeutic effect of MSCs in experimental infarction [ 25 ]. It is known that the cytoprotective effect of stem cells located near damaged cells is due to their paracrine and endocrine activity [ 26 ]. For example, all paracrine factors secreted by SCs in damaged myocardium have been described in detail. Similar examples also exist for lymphoid stem cells (T- and B-cells), which secrete paracrine factors that promote the repair of damaged skeletal muscle [ 27 ], as well as for other types of stem cells that secrete neurotrophic factors in response to brain cell damage [ 28 ] or factors that stimulate the repair of damaged kidneys [ 29 ]. Summarizing all of the above, we can assume the following chain of events: during intercellular contacts, the cytoplasm of neutral cells enters the MSCs, inducing an increase in the production of myotropic factors; then, more effective repair of damaged tissue occurs due to the enhanced paracrine activity of MSCs, and mitochondria are transferred from MSCs to neutral cells damaged by ischemia. Conclusion The experimental rationale included exploring the impact of different administration methods and locations on the effectiveness of cellular cardiomyoplasty for refractory angina. The choice of autologous MSCs and various injection routes allowed for a comprehensive assessment of potential therapeutic outcomes. The results of the stress load modeling in the experimental groups provide valuable insights into the impact of cellular cardiomyoplasty on heart function in the context of myocardial infarction (MI). Here are some key observations based on the information provided: Positive Chronotropic Effect: a positive chronotropic effect, indicating an increase in heart rate (HR), was observed during isopropyl norepinephrine loading in all groups of animals. The extent of this effect varied significantly among the different groups. Baseline HR in Intact Rats: in intact rats (Group 1), the absolute increase in the first minute of isopropyl norepinephrine loading was approximately 32 beats/min. Effects in Group 2 (Empty Injections): Animals in Group 2 that received "empty" myocardial injections demonstrated a stress-induced increase in HR of 11 beats/min. This group had a high baseline HR. Effects in Group 3 (Autologous MSCs into Myocardium): animals in Group 3, treated with autologous mesenchymal stem cells (MSCs) injected into the myocardium, exhibited a stress-induced increase in HR of 22 beats/min. General Trend in HR by the Third Minute: by the third minute of the experiment, a decrease in HR was observed in all groups, with stabilization at 490–495 beats/min. Implications for Myocardial Infarction and Heart Failure: the findings suggest that in the groups with untreated MI and "empty" myocardial injections (Groups 1 and 2), compensation of the required blood flow is deemed impossible despite stress. This lack of compensation aligns with the clinical manifestation of heart failure, emphasizing the importance of addressing myocardial infarction and its consequences. In summary, the study indicates that cellular cardiomyoplasty, particularly with autologous MSCs injected into the myocardium, may positively impact stress-induced changes in heart rate. However, further analysis and consideration of additional cardiovascular parameters are necessary to comprehensively evaluate the potential therapeutic effects of cellular cardiomyoplasty for refractory angina and its relevance to heart failure clinic outcomes. Abbreviations DMEM ECG - Electrocardiography EDTA EF - ejection fraction FS - fractional shortening IHD - Ischemic heart disease HF - Heart failure LVIDd - end-diastolic left ventricular internal dimensions; LVIDs - end-systolic left ventricular internal dimensions; LVEF - left ventricular ejection fraction LVC - left ventricular cavity MSCs - mesenchymal stem cell NO - nitric oxide RA - refractory angina SV - stroke volume. SF - shortening fraction UNOX - number of stable end metabolites of NO, namely NO2¯ + NO3¯ VEGF - vascular endothelial growth factor Declarations Availability of data and materials The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request. Acknowledgements The authors would like to thank the patients and clinical staff who gave up their time to take part in this clinical study. Authors’ contributions IAK, SG, YVI and SIE constructed the study design. IAK, SIE, KS and YVI contributed to data interpretation and manuscript drafting. YVI and SG, contributed to the statistical analysis. KS and SG prepared figures. KS, SG, and YVI participated in the clinical investigation and contributed to the epidemiological data collection. SG and KS revised the manuscript. All authors read and approved the final manuscript. Consent for publication - Not applicable Funding sources This research was part of the research work of the Ethics Committee of Kharkiv National Medical University, Ukraine (Protocol No. 6, November 11, 2022) and Ukraine Association of Biobank (Protocol dd. 11/11/2021) and conducted by the Law of Ukraine "On Protection of Animals from Cruelty" and the provisions of the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes by ARRIVE guidelines. The number of state registration is 0116u00499. Ethical Approval and consent to participate The study within the framework of the academic research work at the basis of the study and the use of data were consented to by the Ethics Committee of Kharkiv National Medical University, Ukraine (Protocol No. 6, November 11, 2022) and Ukraine Association of Biobank (Protocol dd. 11/11/2021) and conducted by the Law of Ukraine "On Protection of Animals from Cruelty" and the provisions of the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes by ARRIVE guidelines. The number of state registration is 0116u00499. Conflicts of Interest All the authors declare no conflicts of interest. Informed consent process Informed consent was obtained from all participants included in the study. Information about authors: Igor A. Kryvoruchko , Ph.D., Doctor of Medical Sciences, Professor, Head of the Department of Surgery No. 2, Kharkiv National Medical University, Kharkiv, Ukraine. email: [email protected] ORCID: https://orcid.org/0000-0002-5525-701X Sergii I. Estrin , Ph.D., Doctor of Medical Sciences, Professor, Head of the Department of Cardio Surgery No. 2, Kharkiv National Medical University, Kharkiv, Ukraine. email: [email protected] ORCID: https://orcid.org/0000-0003-3957-5971 Yulia V. Ivanova , Ph.D., Doctor of Medical Sciences, Professor, Professor of the Department of Surgery No. 1, Kharkiv National Medical University, Kharkiv, Ukraine. email: [email protected] https://orcid.org/0000-0003-4464-3035 Svetlana Gramatiuk , PhD, President Ukraine Association of Biobank, MSc Biobanking Lecturer, International Biobanking and Education, Medical University of Graz, Austria. email: [email protected] https://orcid.org/0000-0003-4238-7031 Karine Sargsyan, Ph.D ., Doctor of Medical Sciences, Professor, Head of the International Biobanking and Education, Medical University of Graz, Austria. email: [email protected] (KS) https://orcid.org/0000-0001-5853-4994 References Rosenzweig A. Cardiac cell therapy-mixed results from mixed cells /. N Engl J Med. 2006;355:1274–87. Qian L, Huang Y, Spencer CI et al. In vivo reprogramming of murine cardiac fibroblasts into induced cardiomyocytes. Nat 2012 Apr 18.P593–8. Psaltis PJ, Carbone A, Nelson AJ, Lau DH, Jantzen T, Manavis J, et al. Reparative effects of allogeneic mesenchymal precursor cells delivered transendocardially in experimental nonischemic cardiomyopathy. JACC Cardiovasc Interv. 2010;3:974–83. Perico N, Casiraghi F, Introna M, Gotti E, Todeschini M, Cavinato RA, et al. Autologous mesenchymal stromal cells and kidney transplantation: A pilot study of safety and clinical feasibility. Clin J Am Soc Nephrol. 2011;6:412–22. Mocini D, Staibano M, Mele L, et al. Autologous bone marrow mononuclear cell transplantation in patients undergoing coronary artery bypass grafting. Am Heart J. 2006;151:192–7. Melzack R, Wall PD. Pain mechanisms: A new theory.Science, 1965; 150: 971–979. Donndorf P, Kaminski A, Tiedemann G, Kundt G, Steinhoff G. Validating intramyocardial bone marrow stem cell therapy in combination with coronary artery bypass grafting, the PERFECT Phase III randomized multicenter trial: study protocol for a randomized controlled trial. Trials. 2012;13:99. Donndorf P, Kundt G, Kaminski A, Yerebakan C, Liebold A, Steinhoff G, Glass A. Intramyocardial bone marrow stem cell transplantation during coronary artery bypass surgery: a meta-analysis. J Thorac Cardiovasc Surg. 2011;142(4):911–20. Efficacy. and safety of Ad5FGF-4 for myocardial ischemia in patients with stable angina due to coronary artery disease (ASPIRE). http://clinicaltrials.gov/ct2/show/NCT01550614?term=gene+therapy+Coronary+Heart+Disease+Phase+3&rank=8 . Gnecchi M, Zhang Z, Ni A et al. Paracrine mechanisms in adult stem cells signaling and therapy // Circ. Res.-2008.-Vol.103.-P.1204–1219. Urbanek K, Rota M, Cascapera S et al. Cardiac stem cells possess growth factor-receptor systems that, after activation, regenerate the infarcted myocardium, improving ventricular function and long-term survival. Circ Res2005;(97):92–4. Traverse JH, Henry TD. Moye' LA. Is the left ventricular ejection fraction measurement the proper endpoint for cell therapy trials? When evaluated by cardiac magnetic resonance imaging, an analysis of the effect of bone marrow mononuclear stem cell administration on left ventricular ejection fraction after ST-segment elevation myocardial infarction. Am Heart J. 2011;162(4):671–7. Hedman M, Muona K, Hedman A, et al. Eight-year safety follow-up of coronary artery disease patients after local intracoronary VEGF gene transfer. Gene Ther. 2009;16(5):629–34. Intracoronary infusion of bone marrow-derived selected CD34 + CXCR4 + cells, Tendera M, Wojakowski W, Ruzyłło W, Chojnowska L, Kepka C, Tracz W, Musiałek P, Piwowarska W, Nessler J, Buszman P, Grajek S, Breborowicz P, Majka M, Ratajczak MZ. non-selected mononuclear cells in patients with acute STEMI and reduced left ventricular ejection fraction: results of randomized, multicentre Myocardial Regeneration by Intracoronary Infusion of Selected Population of Stem Cells in Acute Myocardial Infarction (REGENT) Trial. REGENT Investigators Eur Heart J. 2009;30(11):1313–21. Intramyocardial, autologous CD34 + cell therapy for refractory angina, Losordo DW, Henry TD, Davidson C, Sup Lee J, Costa MA, Bass T, Mendelsohn F, Fortuin FD, Pepine CJ, Traverse JH, Amrani D, Ewenstein BM, Riedel N, Story K, Barker K, Povsic TJ, Harrington RA, Schatz RA. Investigators Circ Res. 2011;109(4):ACT34–CMI. Rosenzweig A. Cardiac cell therapy-mixed results from mixed cells /. N Engl J Med. 2006;355:1274–87. Qian L, Huang Y, Spencer CI et al. In vivo reprogramming of murine cardiac fibroblasts into induced cardiomyocytes. Nat 2012 Apr 18.P593–8. Debayon Paul 1, Samuel SM, Maulik N. Mesenchymal stem cell: present challenges and prospective cellular cardiomyoplasty approaches for myocardial regeneration. Antioxid Redox Signal. 2009;11(8):1841–55. 10.3390/molecules28134914 . Published online 2023 Jun 22. Abdel-Latif A, Bolli R, Zuba-Surma EK, Tleyjeh IM, Hornung CA, Dawn B. Granulocyte colony-stimulating factor therapy for cardiac repair after acute myocardial infarction: a systematic review and meta-analysis of randomized controlled trials. Am Heart J. 2008;156:216–26. e9. Zhang M, Wang Z-Z, Nai-Hong Chen. Connexin 43 Phosphorylation: Implications in Multiple Diseases./ Molecules. 2023; 28(13): 4914. Published online 2023 Jun 22. 10.3390/molecules28134914 . PMCID: PMC10343229. Arminan A, Gandia C, Bartual C, Garcia-Verdugo JM, Lledo E, Mirabet V, Llop M, Barea J, Montero JA, Sepulveda P. Cardiac differentiation is driven by NKX2.5 and GATA4 nuclear translocation in tissue specific mesenchymal stem cells. Stem Cells Dev. 2008. Nov 4 [Epub ahead of print]. Georges Makhoul, Ray CJ, Chiu R, Cecere. Placental mesenchymal stem cells: a unique source for cellular cardiomyoplasty. Ann Thorac Surg. 2013;95(5):1827-33. 10.1016/j.athoracsur.2012.11.053 . Epub 2013 Mar 28. PMID: 23541427 DOI: 10.1016/j.athoracsur.2012.11.053. Malek A, Bersinger NA. Human placental stem cells: biomedical potential and clinical relevance. J Stem Cells. 2011;6(2):75–92. PMID: 22997848. Walther G, Gekas J, Bertrand OF. Amniotic stem cells for cellular cardiomyoplasty: promises and premises. Catheter Cardiovasc Interv. 2009;73(7):917 – 24. 10.1002/ccd.22016 . PMID: 19455667. Pourrajab F, Forouzannia SK, Tabatabaee SA. Molecular characteristics of bone marrow mesenchymal stem cells, source of regenerative medicine. Int J Cardiol. 2013;163(2):125–31. Epub 2011 Dec 2. PMID: 22137091. Shevchenko EK, Dergilev KV, Tsokolaeva ZI, Beloglazova IB, Molokotina YD, Parfenova EV, Men'shikov MY. Combination of Mesenchymal Stromal Cells and Cardiac Stem Cells in a Multilayer Cell Construct Promotes Activation of Notch Signaling and Initiation of Endothelial Differentiation. Bull Exp Biol Med. 2019;166(4):548–52. 10.1007/s10517-019-04390-7 . Epub 2019 Feb 19. PMID: 30783844. Valarmathi MT, Fuseler JW, Davis JM, Price RL. A Novel Human Tissue-Engineered 3-D Functional Vascularized Cardiac Muscle Construct. Front Cell Dev Biol. 2017;5:2. 10.3389/fcell.2017.00002 . eCollection 2017. Preda MB, Rosca AM, Tutuianu R, Burlacu A. Pre-stimulation with FGF-2 increases in vitro functional coupling of mesenchymal stem cells with cardiac cells. Biochem Biophys Res Commun. 2015;464(2):667–73. Epub 2015 Jul 14. PMID: 26187662. Makhoul G, Jurakhan R, Jaiswal PK, Ridwan K, Li L, Selvasandran K, Duong M, Schwertani A, Cecere R. Conditioned medium of H9c2 triggers VEGF dependent angiogenesis by activation of p38/pSTAT3 pathways in placenta derived stem cells for cardiac repair. Life Sci. 2016;153:213 – 21. 10.1016/j.lfs.2016.04.009 . Epub 2016 Apr 16. PMID: 27091377. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4024504","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":277312603,"identity":"8622c1ec-c8ac-422c-8f88-7ae8f8ae2457","order_by":0,"name":"Svetlana Gramatiuk","email":"","orcid":"","institution":"Ukraine Association of Biobank","correspondingAuthor":false,"prefix":"","firstName":"Svetlana","middleName":"","lastName":"Gramatiuk","suffix":""},{"id":277312604,"identity":"5007d8dd-b8ce-4ced-ba49-2fbda5141495","order_by":1,"name":"Yulia Ivanova","email":"","orcid":"","institution":"Kharkiv National Medical University","correspondingAuthor":false,"prefix":"","firstName":"Yulia","middleName":"","lastName":"Ivanova","suffix":""},{"id":277312605,"identity":"6d1224e1-e978-4c0c-8cb6-9bcafc1d5f47","order_by":2,"name":"Karine Sargsyan","email":"data:image/png;base64,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","orcid":"","institution":"Medical University of Graz","correspondingAuthor":true,"prefix":"","firstName":"Karine","middleName":"","lastName":"Sargsyan","suffix":""},{"id":277312606,"identity":"5a777f51-04c8-4ab2-9957-445169e4a7fd","order_by":3,"name":"Igor Kryvoruchko","email":"","orcid":"","institution":"Kharkiv National Medical University","correspondingAuthor":false,"prefix":"","firstName":"Igor","middleName":"","lastName":"Kryvoruchko","suffix":""},{"id":277312607,"identity":"ad235f31-8595-440e-aaa8-df0c86de0064","order_by":4,"name":"Sergii Estrin","email":"","orcid":"","institution":"Kharkiv National Medical University","correspondingAuthor":false,"prefix":"","firstName":"Sergii","middleName":"","lastName":"Estrin","suffix":""}],"badges":[],"createdAt":"2024-03-07 12:45:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4024504/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4024504/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":67800145,"identity":"c1d9e066-05e8-4ad5-9563-b39a3d08890f","added_by":"auto","created_at":"2024-10-29 22:31:31","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":539325,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4024504/v1/9354cb7d-081b-4e38-9e41-4e23285e1bf7.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Cellular cardiomyoplasty for refractory angina: experimental rationale.","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIschemic heart disease (IHD) is the most common cause of cardiovascular disease in terms of complication frequency and number of fatal outcomes. In the United States, it is the cause of every fifth death [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. In Ukraine, the diagnosis of IHD is established for approximately 400,000 patients annually [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Traditional methods of treating this category of patients, which currently exist, are:\u003c/p\u003e \u003cp\u003eThe available treatments for ischemic heart disease include medication, direct myocardial revascularization (coronary artery bypass grafting or angioplasty with stenting), and heart transplantation. However, current medication therapy is often insufficient to prevent myocardial remodeling [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Heart failure (HF) caused by IHD or cardiomyopathies is a severe disease with a poor prognosis.\u003c/p\u003e \u003cp\u003eDespite the wide range of pharmaceuticals and surgical interventions available, there remains a significant number of patients with angina for whom surgery is not possible for various reasons and for whom drug therapy is insufficiently effective [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe definition of refractory angina (RA) was first proposed in 2002 by a joint group of the European Society of Cardiology for treating refractory angina. It is a chronic condition (lasting more than three months) characterized by the presence of angina caused by insufficient coronary blood flow (due to coronary artery disease), accompanied by severe clinical symptoms. Recent studies in stem cell biology have fundamentally changed all concepts of myocardial regenerative capacity and have become the beginning of a new therapeutic direction - cellular cardiomyoplasty. Patients who cannot be controlled with combined drug therapy at the maximum tolerated doses and are unable to undergo myocardial revascularization (percutaneous coronary angioplasty or coronary artery bypass grafting) [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe text discusses stem cell transplantation as a potential therapy for patients with chronic ischemic heart disease. However, no large-scale studies have evaluated the effectiveness of stem cell implantation in patients with heart failure.\u003c/p\u003e \u003cp\u003eMany fundamental issues in cell therapy remain unresolved, including homing mechanisms, differentiation and engraftment of transplanted stem cells, the role of cell fusion, and the mechanisms by which transplanted cells affect the function and metabolism of the heart muscle. The most effective method of delivering cells to the myocardium, the timing of cardiomyoplasty, the number of cells in the transplant, and methods of preparation are also topics of discussion. However, much research has been devoted to these issues [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThis experimental \u003cb\u003estudy aimed\u003c/b\u003e to investigate the therapeutic potential of cellular cardiomyoplasty in the context of refractory angina using a rodent model.\u003c/p\u003e \u003cp\u003eThe study within the framework of the academic research work at the basis of the study and the use of data were consented to by the Ethics Committee of Kharkiv National Medical University, Ukraine (Protocol No. 6, November 11, 2022) and Ukraine Association of Biobank (Protocol dd. 11/11/2021) and conducted by the Law of Ukraine \"On Protection of Animals from Cruelty\" and the provisions of the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes by ARRIVE guidelines.\u003c/p\u003e "},{"header":"Materials and methods","content":"\u003cp\u003eThe experimental part of the work was carried out on 122 rats of the inbred Wistar-Cayote line weighing 200\u0026ndash;220 g, kept in a vivarium in compliance with ARRIVE recommendations. We used the Wistar-Kayota breed because it is an inbred breed that minimizes the rejection reaction, given its genetic homogeneity. The animals were used in the experiment by the rules regulated by the European Convention for the Care and Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes (Strasbourg, 1986), the Council of the European Community Directive of 24.11.86 and the order of the Ministry of Health of Ukraine No. 32 of 22.02.88. Surgical interventions were performed in an experimental operating room under ketamine anesthesia (12.5 mg/100 g of body weight intramuscularly).\u003c/p\u003e \u003cp\u003eMI induction was performed by stitching and ligation of the anterior interventricular artery. Only animals with transmural MI, which ECG and ultrasound confirmed, were included in the further experiment.\u003c/p\u003e \u003cp\u003eThe effect of cellular cardiomyoplasty on the course of MI was studied in 100 surviving animals (22 animals died in the first hours after modeling the pathological condition due to the development of life-threatening arrhythmias). The animals included in the study were withdrawn from the experiment by decapitation (under general anesthesia).\u003c/p\u003e \u003cp\u003eThe animals were divided into five groups (20 animals in each group): in group 1, no treatment was performed; in group 2, \"empty\" injections were performed into the myocardium in the area of the ischemia zone, which was determined macroscopically, in group 3, autologous mesenchymal stem cells (MSCs) were injected at a dose of 10\u0026nbsp;million cells. In group 4, MSCs were injected intravenously at the same dose by puncture of the tail vein; in group 5, MSCs were injected into the left ventricular cavity (LVC) by puncture and catheter through the right femoral artery (this was done to create the maximum concentration of MSCs in the coronary vessel mouth and to simulate intracoronary injection.\u003c/p\u003e \u003cp\u003eMSCs were obtained from the peripheral blood of animals according to the following procedure. A syringe containing 0.5 ml of phosphate-buffered saline, 50 units/ml of heparin, and 0.25 mg/l of gentamicin sulfate was used to collect 0.5 ml of blood from the tail vein of the animal. The volume of blood taken from the animals was acceptable and safe, as it did not exceed 10% of the circulating blood volume (13.75\u0026ndash;17.50 ml for rats, based on the recommendations for safe blood collection in mice and rats of the US National Institutes of Health). The cell suspension was centrifuged at 1500 rpm for 5 min; the cell pellet was resuspended in erythrocyte lysis solution (114 mM ammonium chloride, 7.5 mM bicarbonate, 100 \u0026micro;M EDTA) for 3 min and centrifuged again. The haemolysed supernatant was removed, and the cell pellet was resuspended in DMEM containing 10% fetal calf serum (HyClonegold, USA), insulin 0.4 \u0026micro;M, and 0.25 mg/l gentamicin sulfate. The resulting cells were seeded in culture vials and transferred to a CO\u003csub\u003e2\u003c/sub\u003e incubator with a 5% carbon dioxide concentration and 95% humidity. 2 days after the primary culture was isolated, the non-attached cell suspension was removed, and the remaining cells with fibroblast-like morphology were continued to be cultured. The culture medium was replaced with fresh medium every 3 to 4 days. After 75\u0026ndash;80% monolayer formation, the cells were washed once with Versailles solution, removed with Versailles solution with 0.25% trypsin solution, resuspended in growth medium, and poured into new culture dishes. The cellular material, flattened fibroblast-like cells (mesenchymal stromal cells) fixed on plastic, retained population activity, and did not contain dead cells, was considered suitable for use. The stromal origin of the stem cells was confirmed by immunohistochemical method in culture by detecting type I collagen using rabbit monoclonal antibodies. MSCs of the first and second passages were used for the experiment. The viability of MSCs was determined before their injection by trypan blue staining. For this purpose, a 0.1% trypan blue solution was used for the cell suspension, and after 2\u0026ndash;3 min, live cells were counted in a phase-contrast microscope without staining their membranes.\u003c/p\u003e \u003cp\u003eMarkers of neoangiogenesis in the blood serum were studied in laboratory animals in the dynamics. The nitric oxide (NO) content in the blood plasma was judged by the number of stable end metabolites of NO, namely NO\u003csub\u003e2\u003c/sub\u003e\u0026macr; + NO\u003csub\u003e3\u003c/sub\u003e\u0026macr; (UNOX). The concentration of vascular endothelial growth factor (VEGF) was measured using a Luminex dual-laser flow cytometric fluorescence analyzer (Luminex Corporation, USA) based on Simplex ProcartaPlexTM reagent kits (Affymetrix, USA). The level of endothelin-1 in blood plasma was determined by enzyme-linked immunosorbent assay using the Endotelin (1\u0026ndash;21) kit from Biomedica (Austria) on a Stat Fax 2100 enzyme-linked immunosorbent assay.\u003c/p\u003e \u003cp\u003eStudy period: 1, 6, and 24 hours after induction of the pathological condition and treatment, as well as on days 7 and 30 of the study.\u003c/p\u003e \u003cp\u003eEchocardiography was performed using a GE Vivid newborn ultrasound machine (USA) with a 12 MHz transducer. Study period: 1 and 3 months of the experiment. The following rat heart parameters were studied in different experimental groups: LVIDd - end-diastolic left ventricular internal dimensions; LVIDs - end-systolic left ventricular internal dimensions; FS - fractional shortening; EF - ejection fraction; SV - stroke volume.\u003c/p\u003e \u003cp\u003eElectrocardiography (ECG) was performed on an IVF 1T apparatus on day 30 of the experiment.\u003c/p\u003e \u003cp\u003eUsed the standard statistical software package SPSS 20.0 for Windows for statistical data processing. Statistical data processing was performed using a non-parametric Kruskal-Wallis test and Statistica software (StatSoft, Inc., USA). Considered differences statistically significant at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e "},{"header":"Results of the experimental block of studies","content":" \u003cp\u003eWhen studying the levels of vasoconstrictor endothelin-1, vasodilator nitric oxide, and endothelial vascular growth factor in the dynamics one month after modeling MI, determined a general pattern for all three indicators: they reached their maximum value one day after modeling MI (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e ).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eChanges in the concentration of angiogenesis markers in rat blood serum\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eIndicators\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c7\" namest=\"c3\"\u003e \u003cp\u003eThe frame of the study\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eAnimal groups\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1 hour\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6 hours\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24 hours\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSeven days\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e30 days\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNO\u003c/p\u003e \u003cp\u003e(mg/ml)\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003cp\u003e*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003cp\u003e*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eVEGF\u003c/p\u003e \u003cp\u003e(pg/ml)\u003c/p\u003e \u003cp\u003e( n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e66.98\u0026thinsp;\u0026plusmn;\u0026thinsp;12.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e70.21\u0026plusmn;\u003c/p\u003e \u003cp\u003e11.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e126.72\u0026plusmn;\u003c/p\u003e \u003cp\u003e24.05*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e220.45\u0026plusmn;\u003c/p\u003e \u003cp\u003e22.13*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e134.89\u0026plusmn;\u003c/p\u003e \u003cp\u003e25.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e89.74\u0026plusmn;\u003c/p\u003e \u003cp\u003e21.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e71.42\u0026plusmn;\u003c/p\u003e \u003cp\u003e13.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e134.86\u0026plusmn;\u003c/p\u003e \u003cp\u003e28.11*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e288.22\u0026plusmn;\u003c/p\u003e \u003cp\u003e23.46***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e189.57\u0026plusmn;\u003c/p\u003e \u003cp\u003e28.47*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e132,74\u0026plusmn;\u003c/p\u003e \u003cp\u003e19,87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eEndothelin-1 (fmol/ml)\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e5.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e \u003cp\u003e*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0,5\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e \u003cp\u003e**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eNote: when compared with the previous indicator: * - p\u0026thinsp;\u0026lt;\u0026thinsp;0.05; ** - p\u0026thinsp;\u0026lt;\u0026thinsp;0.01; *** - p\u0026thinsp;\u0026lt;\u0026thinsp;0.001; 1 - untreated MI group; 2 - MI\u0026thinsp;+\u0026thinsp;i.v. MSCs transplantation.\u003c/p\u003e \u003cp\u003eThe experimental data obtained indicated a decrease in myocardial ischemia after MSCs transplantation, which led to a decrease in the level of such a potent vasoconstrictor as endothelin1.\u003c/p\u003e \u003cp\u003eWhen studying the dynamics of VEGF concentration, which reflects the intensity of angiogenesis, it was found that up to 6 hours after MI induction, this indicator did not differ from control values. After 6 hours in the group of animals with MI, the VEGF content increased and reached its maximum peak by the end of the 1st day. By the end of the experiment, the level of VEGF decreased to (89.74\u0026thinsp;\u0026plusmn;\u0026thinsp;21.38) pg/ml, which did not differ from normal values (t\u0026thinsp;=\u0026thinsp;0.91; p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). In the group of animals with MI\u0026thinsp;+\u0026thinsp;MSCs transplantation, a significant increase in VEGF levels was also observed by 6 hours of the study, reaching its maximum by the end of the 1st day; its level decreased slightly and by the end of the experiment was (132.74\u0026thinsp;\u0026plusmn;\u0026thinsp;19.87) pg/ml, which was two times higher than normal values (t\u0026thinsp;=\u0026thinsp;2.8; p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003eWhen analysing the effect of MSCs transplantation on vascular tone parameters after MI, it was found that cellular cardiomyoplasty had a pronounced vasodilator effect from the first day. This was reflected in an increase in NO levels with a simultaneous decrease in the content of the vasoconstrictor endothelin 1. It is characteristic that the increased nitric oxide content persisted until the experiment's end.\u003c/p\u003e \u003cp\u003eWhen studying the concentrations of nitric oxide derivatives in groups of animals with different methods of MSCs administration, it found that after one month in rats of the 1st and 2nd groups, their increase to (0.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03) and (0.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02) \u0026micro;g/ml, respectively, occurred (t\u0026thinsp;=\u0026thinsp;6.4, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). In the 3rd, 4th, and 5th groups, a more significant increase in the studied indicator was found: (0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04), (0.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05) and (0.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03) \u0026micro;g / ml, respectively, while in the 3rd group, the NO content was higher than in the 5th group (t\u0026thinsp;=\u0026thinsp;1.99, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eConcentrations of angiogenesis markers in rat blood serum 30 days after MI modeling\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eIndicators\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c7\" namest=\"c3\"\u003e \u003cp\u003eAnimal groups\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1st (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2nd (n\u0026thinsp;=\u0026thinsp;18)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3rd (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4th (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5th (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNO\u003c/p\u003e \u003cp\u003e(mg/ml)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.85\u0026plusmn; 0.02***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e \u003cp\u003e0.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 ***\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVEGF\u003c/p\u003e \u003cp\u003e(pg/ml)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e66.98\u0026thinsp;\u0026plusmn;\u0026thinsp;12.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.85\u0026plusmn; 0.02***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e \u003cp\u003e0.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 ***\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEndothelin-1\u003c/p\u003e \u003cp\u003e(fmol/ml)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e5.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e5.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e4.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e \u003cp\u003e5.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eNote: in comparison with the control: * - p\u0026thinsp;\u0026lt;\u0026thinsp;0.05; ** - p\u0026thinsp;\u0026lt;\u0026thinsp;0.01; *** - p\u0026thinsp;\u0026lt;\u0026thinsp;0.001.\u003c/p\u003e \u003cp\u003eA similar trend was observed concerning the concentration of VEGF: in groups 1st and 2nd, its level did not exceed average values; in groups 3rd, 4th and 5th, the studied indicator was significantly higher than usual and amounted to (144.22\u0026thinsp;\u0026plusmn;\u0026thinsp;14.59), (132.74\u0026thinsp;\u0026plusmn;\u0026thinsp;19.87) and (111.43\u0026thinsp;\u0026plusmn;\u0026thinsp;12.22) pg/ml, respectively, while, by analogy with the level of nitric oxide, determined the maximum level in group 3 compared to animals in group 5 (t\u0026thinsp;=\u0026thinsp;1.97, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003eAnalysis of the concentration of endothelin 1 showed that in the 3rd, 4th, and 5th groups one month after the experiment's induction, the level of the studied indicator did not differ from the norm. In the 1st and 2nd groups, it was slightly higher than normal values and amounted to (5.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2) and (5.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2) fmol/ml, respectively.\u003c/p\u003e \u003cp\u003eWhen studying the final diastolic LV internal diameter, it was found that in the 1st group of animals, it increased from (6.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12) to (7.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16) mm, t\u0026thinsp;=\u0026thinsp;5.15, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001. A similar trend was determined in the second group: this indicator increased to (7.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14) mm, t\u0026thinsp;=\u0026thinsp;5.7, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001. In animals of groups 3 and 4, LVIDd remained unchanged at t\u0026thinsp;=\u0026thinsp;0.12 and 0.98, respectively. In group 5, LVIDd also increased to (6.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15) mm at t\u0026thinsp;=\u0026thinsp;3.4, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01. In the study of LVIDs, it was found that in the 1st group of rats, it increased from (2.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18) to (3.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11) mm at t\u0026thinsp;=\u0026thinsp;4.7, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. In animals of the 2nd group, this indicator increased to (3.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13) mm at t\u0026thinsp;=\u0026thinsp;4.2, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01. It should be noted that in the 3rd and 4th groups, LVIDs remained unchanged, and in the 5th group, it increased to (3.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13) mm at t\u0026thinsp;=\u0026thinsp;3.5, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. At the same time, LVIDs in group 5th did not differ significantly from those recorded in rats of groups 1st and 2nd and were higher than in group 3rd at t\u0026thinsp;=\u0026thinsp;4.0, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01. Thus, the best values of LVIDd and LVIDs were obtained in animals of groups 3rd and 4th (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003cb\u003eUltrasound parameters of heart function in rats\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eIndicators\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e \u003cp\u003eAnimal groups\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1st (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2nd (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3rd (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4th (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5th (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLVIDd, mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e6.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e7.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e7.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e6.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e6.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e \u003cp\u003e6.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLVIDs, mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e2.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e3.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e3.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e2.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e2.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e \u003cp\u003e3.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSF, %\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e45.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e26.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e27.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e45.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e42.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e \u003cp\u003e31.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLVEF, %\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e76.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e55.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e56.2\u0026thinsp;\u0026plusmn;\u0026thinsp;3.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e75.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e70.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e \u003cp\u003e66.5\u0026thinsp;\u0026plusmn;\u0026thinsp;3.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSV, ml\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e0.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e0.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e \u003cp\u003e0.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eNote: The statistical significance of the figures is given in the text.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eWhen comparing the shortening fraction (SF), it was found that in animals of the 1st and 2nd groups, this indicator decreased by (26.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8) and (27.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7)%, respectively, at t\u0026thinsp;=\u0026thinsp;9.8, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001. In group 3rd, it remained unchanged and was equal to (45.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9)% at t\u0026thinsp;=\u0026thinsp;0.16, p\u0026thinsp;\u0026gt;\u0026thinsp;0.05. In group 4th, the SF was slightly lower than normal and was equal to (42.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12)% at t\u0026thinsp;=\u0026thinsp;1.98, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. In the 5th group of animals, the SF was equal to (31.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9)%, below typical values at t\u0026thinsp;=\u0026thinsp;7.3, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001. At the same time, the studied index was higher in group 5th compared to the results obtained in animals of groups 1st and 2nd at t\u0026thinsp;=\u0026thinsp;3.7 and 3.4, respectively, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, and higher than in groups 3rd and 4th at t\u0026thinsp;=\u0026thinsp;10.8 and 12.6, respectively, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001. In addition, in animals of group 3rd, SF was higher than in group 4 at t\u0026thinsp;=\u0026thinsp;2.6, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e \u003cp\u003eA similar trend was observed in the analysis of changes in left ventricular ejection fraction (LVEF) dynamics. Thus, in group 1st, it decreased from (76.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5) to (55.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.4)% at t\u0026thinsp;=\u0026thinsp;5.1, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. In the 2nd group, the same picture was observed, with no difference in the indicators between groups 1st and 2nd. In animals of group 3rd, LVEF did not differ from the norm, and in group 4th, this indicator was below the norm and was equal to (70.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3)% at t\u0026thinsp;=\u0026thinsp;1.9, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. In animals of group 5th, LVEF also decreased to (66.5\u0026thinsp;\u0026plusmn;\u0026thinsp;3.3) at t\u0026thinsp;=\u0026thinsp;2.5, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. It should be noted that the LVEF in group 5th was higher than in groups 1st and 2nd at t\u0026thinsp;=\u0026thinsp;2.4 and 2.2, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, but lower than in group 3rd at t\u0026thinsp;=\u0026thinsp;2.02, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. The LVEF between the 4th and 5th groups did not differ significantly.\u003c/p\u003e \u003cp\u003eThe dynamics of changes in stroke volume in the experimental groups were as follows: in group 1st, SV decreased from (0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08) to (0.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03) ml at t\u0026thinsp;=\u0026thinsp;1.9, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, and in the other groups this indicator remained unchanged. However, in group 3rd, the SV was closest to normal and was higher than in groups 1st, 2nd, and 5th of laboratory animals. Thus, the values obtained in groups 3rd, 4th, and 5th animals were the closest to normal, while in group 3rd, they practically did not differ from the norm.\u003c/p\u003e \u003cp\u003eWhen modeling stress load in animals of all groups, a positive chronotropic effect was observed, with the severity of the latter differing significantly in animals of different groups. Thus, in intact rats, the absolute increase in the first minute of isopropyl norepinephrine loading was about 32 beats/min. Against the background of a high baseline heart rate (HR) in animals of the second group, the stress-induced increase was 11 beats/min. In the animals of the third group, the same indicator was 22 beats/min. By the third minute of the experiment, a decrease in HR was observed in all groups of animals with stabilization at 490\u0026ndash;495 beats/min (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eIndicators of the chronotropic function of the corpus callosum under stress modeling of loading\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnimal groups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOutput HR\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMax HR\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eIncrease in HR\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eFinal HR\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e485\u0026thinsp;\u0026plusmn;\u0026thinsp;43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e517\u0026thinsp;\u0026plusmn;\u0026thinsp;35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u0026thinsp;32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e490\u0026thinsp;\u0026plusmn;\u0026thinsp;23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1st (n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e503\u0026thinsp;\u0026plusmn;\u0026thinsp;23\u003cb\u003e*\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e525\u0026thinsp;\u0026plusmn;\u0026thinsp;15\u003cb\u003e*\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u0026thinsp;22\u003cb\u003e*\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e495\u0026thinsp;\u0026plusmn;\u0026thinsp;12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2nd (n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e507\u0026thinsp;\u0026plusmn;\u0026thinsp;18\u003cb\u003e*\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e522\u0026thinsp;\u0026plusmn;\u0026thinsp;11 \u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u0026thinsp;24 \u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e510\u0026thinsp;\u0026plusmn;\u0026thinsp;13\u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3rd (n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e489\u0026thinsp;\u0026plusmn;\u0026thinsp;11\u003cb\u003e*\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e526\u0026thinsp;\u0026plusmn;\u0026thinsp;8 \u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u0026thinsp;38\u003cb\u003e* *\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e494\u0026thinsp;\u0026plusmn;\u0026thinsp;10\u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4th (n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e482\u0026thinsp;\u0026plusmn;\u0026thinsp;14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e528\u0026thinsp;\u0026plusmn;\u0026thinsp;9 \u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u0026thinsp;39\u003cb\u003e* *\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e499\u0026thinsp;\u0026plusmn;\u0026thinsp;15 \u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5th (n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e501\u0026thinsp;\u0026plusmn;\u0026thinsp;16 \u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e527\u0026thinsp;\u0026plusmn;\u0026thinsp;7 \u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u0026thinsp;28\u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e503\u0026thinsp;\u0026plusmn;\u0026thinsp;11\u003cb\u003e**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eNote: * - significance of differences between the studied indicator and the previous data (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05); ** - significance of differences between the studied indicator and the control (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe qualitative response of the heart to the stress-simulating load in different groups of animals was the same; however, at a rate of +\u0026thinsp;32 beats/min, the maximum increase was closest to the norm in groups 3rd, 4th, and 5th, i.e., by +\u0026thinsp;38, +39 and +\u0026thinsp;28 bears/min, respectively. The increase in groups 1st and 2nd was only\u0026thinsp;+\u0026thinsp;22 and +\u0026thinsp;24 beats/min. Thus, in the groups of rats with untreated MI and \"empty\" myocardial injections, compensation of the required blood flow is impossible, despite the stress, which is what the clinic of heart failure is directed to.\u003c/p\u003e"},{"header":"Discussion","content":" \u003cp\u003eIn the current literature, the main question remains open: the mechanism of the protective effect of stem/progenitor cells. Multipotent mesenchymal stromal cells are one of the most attractive cell types for cell therapy due to their proven cardioprotective properties and low immunogenicity [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo date, stem/progenitor cell researchers have been divided into two groups: those who believe that after being introduced into the body, stem cells and progenitor cells differentiate and replace dead or damaged cells and those who think that the paracrine activity of stem cells and progenitor cells, i.e., the synthesis and secretion of specific signaling molecules, is the key to the therapeutic effect of these cells [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Both groups of scientists recognize the positive therapeutic impact of introducing stem or progenitor cells in treating pathological conditions of various organs. The study of protective effects in several studies conducted in rats has shown that introducing MSCs after experimental infarction leads to decreased volume and improved myocardium functional recovery [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIt is recognized that MSCs can promote tissue regeneration by secreting growth factors and other cytokines (including BDNF, NGF, VEGF, and others) [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. In addition, they can regulate the inflammatory response, which plays a vital role in postischemic myocardial damage, as SCs secrete several immunomodulatory factors, such as prostaglandin E2, TNF-a, TGF-b, IL6, and others [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. However, the mechanisms underlying the regenerative potential of MSCs still need to be fully understood and require further study.\u003c/p\u003e \u003cp\u003eOne of the ways to study the mechanisms of SCs' influence on organ function is to directly examine the interaction between the body's cells and transplanted cells to search for chemical compounds that stem cells can secrete into the intercellular space after they are co-cultured with differentiated cells. The same applies to recipient cells, as they can respond to the presence of transplanted cells by synthesizing and secreting various biological compounds. Such \"communication\" between neighboring cells or distant organs within the same organism is very likely to occur. A relatively large amount of data has been obtained confirming the existence of this phenomenon [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. However, intercellular communication as a result of the secretion of biologically active compounds and their \"trapping\" by neighboring cells is limited by diffusion. This limitation can be minimized in the case of the directed transfer of cellular components from cell to cell as a result of the formation of intercellular contacts. Examples of such contacts include the gap junction formed by the connexin 43 protein, which provides intercellular communication, electrical coupling of neighboring cells, and transmission of depolarisation signals [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Among the intercellular interactions, the migration of cytoplasmic components between contacting cells along cell processes, particularly tunneling nanotubes, deserves special attention [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. For several differentiated cells in direct contact with progenitor cells, the exchange of cytosolic components [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e] and intercellular transport of mitochondria [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] is shown. Direct intercellular contact provides the least costly way of exchanging chemical/biological information between cells; however, the efficiency of such exchange is limited by the meager ratio of stem cells to differentiated cells and, as a result, a low number of intercellular contacts. A more costly method of information exchange for cells is the secretion of biologically active molecules [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] into the intercellular space (into the culture medium in vitro and into the extracellular space/interstitial fluid in vivo). At a higher level, communication between cells in different organs can occur. In this case, certain factors are secreted into the bloodstream, which then enters distant organs via the bloodstream.\u003c/p\u003e \u003cp\u003eThus, cellular cardiomyoplasty improves cell metabolism, confirming the neoangiogenesis theory and the paracrine effect of cell transplantation. Therefore, MSC transplantation can optimize cardiomyocyte oxygen metabolism and stabilize aerobic and anaerobic glycolysis. Mitochondrial transport, rather than engraftment of allogeneic MSCs into tissues, can partially explain the positive therapeutic effect of MSCs in experimental infarction [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. It is known that the cytoprotective effect of stem cells located near damaged cells is due to their paracrine and endocrine activity [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. For example, all paracrine factors secreted by SCs in damaged myocardium have been described in detail. Similar examples also exist for lymphoid stem cells (T- and B-cells), which secrete paracrine factors that promote the repair of damaged skeletal muscle [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], as well as for other types of stem cells that secrete neurotrophic factors in response to brain cell damage [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e] or factors that stimulate the repair of damaged kidneys [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Summarizing all of the above, we can assume the following chain of events: during intercellular contacts, the cytoplasm of neutral cells enters the MSCs, inducing an increase in the production of myotropic factors; then, more effective repair of damaged tissue occurs due to the enhanced paracrine activity of MSCs, and mitochondria are transferred from MSCs to neutral cells damaged by ischemia.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe experimental rationale included exploring the impact of different administration methods and locations on the effectiveness of cellular cardiomyoplasty for refractory angina. The choice of autologous MSCs and various injection routes allowed for a comprehensive assessment of potential therapeutic outcomes. The results of the stress load modeling in the experimental groups provide valuable insights into the impact of cellular cardiomyoplasty on heart function in the context of myocardial infarction (MI). Here are some key observations based on the information provided:\u003c/p\u003e \u003cp\u003e \u003col\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003ePositive Chronotropic Effect: a positive chronotropic effect, indicating an increase in heart rate (HR), was observed during isopropyl norepinephrine loading in all groups of animals. The extent of this effect varied significantly among the different groups.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eBaseline HR in Intact Rats: in intact rats (Group 1), the absolute increase in the first minute of isopropyl norepinephrine loading was approximately 32 beats/min.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eEffects in Group 2 (Empty Injections): Animals in Group 2 that received \"empty\" myocardial injections demonstrated a stress-induced increase in HR of 11 beats/min. This group had a high baseline HR.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eEffects in Group 3 (Autologous MSCs into Myocardium): animals in Group 3, treated with autologous mesenchymal stem cells (MSCs) injected into the myocardium, exhibited a stress-induced increase in HR of 22 beats/min.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eGeneral Trend in HR by the Third Minute: by the third minute of the experiment, a decrease in HR was observed in all groups, with stabilization at 490\u0026ndash;495 beats/min.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eImplications for Myocardial Infarction and Heart Failure: the findings suggest that in the groups with untreated MI and \"empty\" myocardial injections (Groups 1 and 2), compensation of the required blood flow is deemed impossible despite stress.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003c/ol\u003e \u003c/p\u003e \u003cp\u003eThis lack of compensation aligns with the clinical manifestation of heart failure, emphasizing the importance of addressing myocardial infarction and its consequences.\u003c/p\u003e \u003cp\u003eIn summary, the study indicates that cellular cardiomyoplasty, particularly with autologous MSCs injected into the myocardium, may positively impact stress-induced changes in heart rate. However, further analysis and consideration of additional cardiovascular parameters are necessary to comprehensively evaluate the potential therapeutic effects of cellular cardiomyoplasty for refractory angina and its relevance to heart failure clinic outcomes.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eDMEM\u003c/p\u003e\n\u003cp\u003eECG - \u0026nbsp;Electrocardiography\u003c/p\u003e\n\u003cp\u003eEDTA\u003c/p\u003e\n\u003cp\u003eEF - ejection fraction\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFS - fractional shortening\u003c/p\u003e\n\u003cp\u003eIHD - \u0026nbsp;Ischemic heart disease\u003c/p\u003e\n\u003cp\u003eHF - Heart failure\u003c/p\u003e\n\u003cp\u003eLVIDd - end-diastolic left ventricular internal dimensions;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eLVIDs - end-systolic left ventricular internal dimensions;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eLVEF - left ventricular ejection fraction\u003c/p\u003e\n\u003cp\u003eLVC - left ventricular cavity\u003c/p\u003e\n\u003cp\u003eMSCs - mesenchymal stem cell\u003c/p\u003e\n\u003cp\u003eNO - nitric oxide\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eRA - refractory angina\u003c/p\u003e\n\u003cp\u003eSV - stroke volume.\u003c/p\u003e\n\u003cp\u003eSF - shortening fraction\u003c/p\u003e\n\u003cp\u003eUNOX - number of stable end metabolites of NO, namely NO2\u0026macr; + NO3\u0026macr;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eVEGF - vascular endothelial growth factor\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the patients and clinical staff who gave up their time to take part in this clinical study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIAK, SG, YVI and SIE constructed the study design. IAK, SIE, KS and YVI contributed to data interpretation and manuscript drafting. YVI and SG, contributed to the statistical analysis. KS and SG prepared figures.\u003c/p\u003e\n\u003cp\u003eKS, SG, and YVI participated in the clinical investigation and contributed to the epidemiological data collection. SG and KS revised the manuscript.\u003c/p\u003e\n\u003cp\u003eAll authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e- Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding sources\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was part of the research work of the Ethics Committee of Kharkiv National Medical University, Ukraine (Protocol No. 6, November 11, 2022) and Ukraine Association of Biobank (Protocol dd. 11/11/2021) and conducted by the Law of Ukraine \"On Protection of Animals from Cruelty\" and the provisions of the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes by ARRIVE guidelines. The number of state registration is 0116u00499.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study within the framework of the academic research work at the basis of the study and the use of data were consented to by the Ethics Committee of Kharkiv National Medical University, Ukraine (Protocol No. 6, November 11, 2022) and Ukraine Association of Biobank (Protocol dd. 11/11/2021) and conducted by the Law of Ukraine \"On Protection of Animals from Cruelty\" and the provisions of the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes by ARRIVE guidelines. The number of state registration is 0116u00499.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the authors declare no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed consent process\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInformed consent was obtained from all participants included in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformation about authors:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eIgor A. Kryvoruchko\u003c/em\u003e\u003c/strong\u003e, Ph.D., Doctor of Medical Sciences, Professor, Head of the Department of Surgery No. 2, Kharkiv National Medical University, Kharkiv, Ukraine.\u003c/p\u003e\n\u003cp\u003eemail: [email protected]\u003c/p\u003e\n\u003cp\u003eORCID: https://orcid.org/0000-0002-5525-701X\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eSergii I. Estrin\u003c/em\u003e\u003c/strong\u003e, Ph.D., Doctor of Medical Sciences, Professor, Head of the Department of Cardio Surgery No. 2, Kharkiv National Medical University, Kharkiv, Ukraine.\u003c/p\u003e\n\u003cp\u003eemail: [email protected]\u003c/p\u003e\n\u003cp\u003eORCID: https://orcid.org/0000-0003-3957-5971\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eYulia V. Ivanova\u003c/em\u003e\u003c/strong\u003e, Ph.D., Doctor of Medical Sciences, Professor, Professor of the Department of Surgery No. 1, Kharkiv National Medical University, Kharkiv, Ukraine.\u003c/p\u003e\n\u003cp\u003eemail: [email protected]\u003c/p\u003e\n\u003cp\u003ehttps://orcid.org/0000-0003-4464-3035\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eSvetlana Gramatiuk\u003c/em\u003e\u003c/strong\u003e, PhD, President Ukraine Association of Biobank, MSc Biobanking Lecturer, International Biobanking and Education, Medical University of Graz, Austria.\u003c/p\u003e\n\u003cp\u003eemail: [email protected]\u003c/p\u003e\n\u003cp\u003ehttps://orcid.org/0000-0003-4238-7031\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eKarine Sargsyan, Ph.D\u003c/em\u003e\u003c/strong\u003e., Doctor of Medical Sciences, Professor, Head of the International Biobanking and Education, Medical University of Graz, Austria.\u003c/p\u003e\n\u003cp\u003eemail: [email protected] (KS)\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;https://orcid.org/0000-0001-5853-4994\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eRosenzweig A. 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PMID: 27091377.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"cellular cardiomyoplasty, autologous mesenchymal stem cell, cardiovascular parameters","lastPublishedDoi":"10.21203/rs.3.rs-4024504/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4024504/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis experimental study aimed to investigate the therapeutic potential of cellular cardiomyoplasty in the context of refractory angina using a rodent model. The study comprised five groups of animals subjected to different interventions, including \"empty\" injections, autologous mesenchymal stem cell (MSC) injections via various routes, and untreated controls. Stress load modeling was employed to assess the hemodynamic response, mainly focusing on heart rate changes during isopropyl norepinephrine loading.\u003c/p\u003e\n\u003cp\u003eResults revealed a positive chronotropic effect in all groups, with varying severity. Intact rats exhibited a significant increase in heart rate, while animals in the \"empty\" injection group demonstrated a less pronounced response. Interestingly, those treated with MSCs, either through direct myocardial injection or intravenous and intracavitary routes, exhibited notable variations in stress-induced heart rate changes. By the third minute of the experiment, a decrease in heart rate was observed across all groups, stabilizing at 490–495 beats/min. Notably, the groups with untreated myocardial infarction and \"empty\" injections displayed an inability to compensate for required blood flow during stress, indicative of potential challenges in these conditions.\u003c/p\u003e\n\u003cp\u003eThese findings suggest that cellular cardiomyoplasty, particularly with autologous MSCs injected into the myocardium, may influence the stress-induced changes in heart rate. However, a comprehensive evaluation of additional cardiovascular parameters and further exploration of therapeutic outcomes are warranted to elucidate the potential clinical relevance of these observations. This study contributes valuable insights into the experimental rationale and hemodynamic effects of cellular cardiomyoplasty, laying the groundwork for future investigations in refractory angina and myocardial infarction.\u003c/p\u003e","manuscriptTitle":"Cellular cardiomyoplasty for refractory angina: experimental rationale.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-12 15:57:18","doi":"10.21203/rs.3.rs-4024504/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"fb522de1-7618-4384-92a9-9afe9c175e55","owner":[],"postedDate":"March 12th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-10-29T22:23:20+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-12 15:57:18","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4024504","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4024504","identity":"rs-4024504","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}