Введение. Вынужденная физическая нагрузка часто нарушает слаженность взаимодействий между корой головного мозга и внутренними органами. Аллогенный биоматериал (БМА) применяется в качестве стимулятора регенерации при его местном применении. Механизм фармакупунктурной коррекции патологических изменений посредством БМА в неокортексе изучен недостаточно. Цель исследования – изучение структуры нервной ткани коры головного мозга в условиях акупунктурного воздействия на биологически активные точки и фармакупунктурной коррекции с БМА. Методика. Моделью анаэробной физической нагрузки явилось принудительное плавание крыс самцов с грузом 10% от массы тела. После проведения плавательного теста в опытной группе (n=20) вводили суспензию БМА акупунктурно, в контрольной (n=20) вводили физиологический раствор. Материал для морфофункционального исследования брали через 5 и 21 сут после принудительной анаэробной физической нагрузки. Результаты. В контрольной группе обнаруживался реактивный глиоз, отек нейропиля, перинуклеарных и периваскулярных пространств, редукция синаптического аппарата, усиление хроматолиза нейроцитов, снижение уровня ингибитора апоптоза Bcl-2+ в клетках. В опытной группе наблюдались признаки восстановления архитектоники слоев нервных клеток неокортекса, увеличения численности синапсов, микроглиальных клеток (CD-68+), Bcl-2+ клеток, снижение количества клеток теней и GFAP+ клеток, восстановление нейроваскулярной единицы, обеспечивающей работу гематоэнцефалического барьера. Заключение. В контрольной группе происходили деструктивные изменения необратимого характера. Акупунктурное воздействие БМА стимулировало нейропротекторные свойства. Introduction. Forced physical activity often disrupts the interactions between the cerebral cortex and internal organs. Allogeneic biomaterial (BMA) is used as a regeneration stimulator when applied topically. The mechanism of pharmacopuncture correction of pathological changes in the neocortex with BMA is not well understood. Aim. To study the structure of nervous tissue in the cerebral cortex after acupuncture of biologically active points and pharma-copuncture administration of BMA. Methods. Anaerobic physical activity was modeled by forced swimming of male rats with a load of 10% of body weight. After the swimming test, in the experimental group (n=20), a BMA suspension was administered by acupuncture. In the control group (n=20), saline was administered. Five and 21 days following the forced anaerobic exercise, tissue was sampled, and morpho-functional studies were performed. Results. In the control group, reactive gliosis, edema of the neuropil, perinuclear and perivascular spaces, reduction of the synaptic apparatus, increased chromatolysis of neurocytes, and a decrease in the apoptosis inhibitor Bcl-2+ in cells were found. In the experimental group, there were signs of restoration of the architectonics of the layers of neocortical nerve cells, an increase in the number of synapses, microglial cells (CD-68+), cell Bcl-2+, a decrease in the number of shadow cells and cell GFAP+, and restoration of the neurovascular unit that ensures the blood-brain barrier functioning. Conclusion. In the brain of control rats, irreversible destructive changes prevailed. Acupuncture of BMA stimulated neuroprotection.

Injectable allogeneic decellularized biomaterials are being developed both as scaffolds for delivery of cellular products and as independent pharmacological agents that affect the cascade of tissue reactions during the period of post-ischemic myocardial remodeling. Biomaterial degradation products can affect cellular processes and modulate cytokine effects, thus determining the healing strategy of damaged tissue. In this work, the influence of biomaterial on the expression of key fibrogenic factors by the cells of tissue bed was demonstrated, and the degree of damage to the myocardium during its ischemic damage was experimentally determined. The aim of our study was to determine the area of myocardial scar degeneration and detection of key fibrogenic factors (bFGF-1, TGFb1, MMP-9), as well as TIMP-2 (MMP-9 antagonist) at the acute and subacute stages of myocardial infarction after implantation of allogeneic powder-like biomaterial in an experimental model. In the course of experiments, the left ventricular coronary artery was ligated in male Wistar rats (experimental group). All animals were divided into 3 groups: experimental group I (n = 50), experimental group II (n = 50), and controls (n = 50). In experimental group I, the artery ligation was simultaneously accompanied by intramyocardial administration of powder-like biomaterial suspension (2 mg). In experimental group II, the allogeneic powder-like biomaterial was administered 5 days after coronary occlusion, and only physiological saline was administered in the control group. The animals were withdrawn from experiment on days +3, +7, +14, +30, and +45. Standard histological assessment (hematoxylin and eosin staining, according to Mallory) and immunohistochemical examination (MMP-9, TGFb1, bFGF-1, TIMP-2) were made, and statistical evaluation was performed. The cells with positive staining were counted, and the scar area index was calculated. We have found that administration of dispersed allogeneic biomaterial was followed by a five-fold decrease in the degree of scar degeneration in both experimental groups at the acute and subacute stages of ischemic myocardial damage as compared to the control group. A significantly decreased expression of fibrogenic factors (MMP-9, TGFb1, bFGF-1) by the local cells was found, along with increased activity of metalloproteinase inhibitor (TIMP-2) in connective tissue cells. Decellularized allogeneic powder-like biomaterial serves as a fibrosis inhibitor and promotes cardioprotection during myocardial remodeling at the initial stages after ischemic injury.

Introduction. The question of the possibility of recovery of postischemic myocardium remains relevant.Aim. The aim of the study was to study the effect of dispersed decellularized allogeneic extracellular matrix (allogeneic biomaterial, DAB) on the developed fibrous degeneration of the myocardium, as well as to reveal the possible mechanisms of cellular regeneration.Materials and methods. The muscular wall of the heart of rats was subjected to cryodestruction. After 45 days, the rats of the main group were intramyocardially injected with a suspension of allogeneic biomaterialinto the area of the affected myocardium, and the rats of the control group were injected with saline.Results and discussions. In the experimental group, there was a regression of the formed fibrous connective tissue, chemoattraction of progenitor cells, their differentiation and integration into the myocardium. The thickness of the muscular part of the wall of the left ventricle was three orders of magnitude higher than in the control group.Conclusion. Analysis of the results of the study indicates that the heart in adult mammals has a powerful regenerative reserve. It is likely that, based on the use of DAB, a protocol can be developed that allows the restoration of the heart muscle even in conditions of already developed fibrous degeneration.

Exhausting physical activity leads to sarcomere overstrain, destruction of the cell membrane, hydrolysis of structural proteins, thus, resulting in irreversible damage to muscle fibers. Allogeneic biomaterial (AB) is applied to regenerate various tissues and organs. The aim of the study was to identify morphofunctional features of the skeletal muscle tissue after physical exertion and under AB application. Material and methods. This experimental study involved male Wistar rats. Anaerobic physical exertion was simulated using the Porsolt test with a load equal 10 % of the body weight for 30 days. After training, animals of the main group (n = 10) were injected AB suspension: 4 ml of a 0.2 % solution into the muscles of the fore and hind limbs totally. Animals of the control group (n = 10) received physiological saline into similar zones. Tolerant load was investigated in 5 and 21 days after injection. Then the animals were withdrawn from the experiment, and muscles of the hind limbs were studied histologically, the total number, the average cross-sectional area of muscle fibers, and the number of necrotic fibers were measured. Results. In 5 and 21 days animals from the control group manifested dystrophic changes in muscle fibers: type III, IV contractures, microcirculation disorders, mosaic necrosis of muscle fibers, inflammatory cell infiltration, and a decreased tolerant load. In 21 days fibrosis was detected. The AB introduction provided rhabdomyogenesis as soon as in 5 days. Inflammatory cell infiltration decreased, the polygonality of muscle fiber profiles was restored, and edematous phenomena were leveled. There was hyperplasia of muscle fibers, a decreased number of necrotic muscle fibers, inhibited fibrosis, and an increased tolerant load. AB further biodegraded. Conclusion. AB contributed to the reduced manifestations of dystrophic changes in muscle fibers, strengthened actoprotective mechanism, and restored physical activity in the early stages.

The aim of research was to study morphological features of the synovial membrane and articular cartilage of rats under simulated adjuvant arthritis and application of allogeneic biomaterial. Material and methods . To simulate rheumatoid arthritis, 12 white outbred female rats were injected 100 μl of complete Freund's adjuvant into the plantar surface of the hind paw. In 7 days, 5 mg of the allogeneic biomaterial “Regeneration stimulator” dissolved in physiological solution was injected into the periarticular bursae of the right knee joint and surrounding tissues in the rats of the main experimental group (n=6). Saline solution was injected into animals of the main control group (n=6). Intact rats (n=6) were included in the intact control group. All the animals were withdrawn from the experiment in 37 days after the onset of the experiment. The knee joints of rats were examined using standard histological, morphometric and statistical methods. Results . Animals of the main control group manifested signs of degenerative changes of varying degrees in the articular apparatus. Proliferation of the synovial cells and tissue hyperplasia were detected in the synovial membrane, these leading to the formation of pannus. There was disorganization of the connective tissue: they were thinned, the fibrous layer was separated, the cartilage and bone tissue were destructed. In animals of the experimental group, there were no signs of tissue hyperplasia and edema, pronounced pannus were not detected after the introduction of the allogeneic biomaterial in the synovial membrane. Mild signs of arthritis manifested as slightly increased processes of the synovial cell proliferation and moderate perivascular infiltration with cellular elements were detected in 2 rats. There were no changes in the structure of the cartilage and bone in the knee joint. Conclusion . Intraarticular injection of the allogeneic biomaterial "Regeneration Stimulator" in rats with simulated autoimmune rheumatoid arthritis is accompanied by a decreased external manifestations of a generalized inflammatory process in the knee joints, and by a decreased inflammation and degenerative changes in the articular apparatus at the tissue level. Notably, the structures of the synovial membrane, the articular cartilage of the knee joint and the underlying subchondral bone are preserved compared with animals of the control group.

Введение. Современные технологии восстановления поврежденного миокарда после инфаркта недостаточно эффективны и не отвечают требованиям полноценной реабилитации больных. Аллогенный биоматериал является стимулятором регенерации мягких тканей и применяется в различных областях медицины: в хирургии, травматологии, офтальмологии, и др. Цель работы - исследование стимуляции регенерации ишемически поврежденного миокарда с помощью биоматериала Аллоплант (БМА) в эксперименте. Методика. Экспериментальные исследования были проведены на 100 крысах-самцах Вистар массой 0,18 - 0,25 кг. Всем животным была проведена коронароокклюзия путем лигирования коронарной артерии. В опытной группе одновременно с стенозированием сосуда интрамиокардиально в бассейн стенозированной артерии вводили суспензию аллогенного биоматериала в количестве 12 мг в физиологическом растворе. В контрольной группе животным вводили физиологический раствор. Взятие материала для исследования проводили через 3, 7, 14, 30 и 45 сут. В работе использовали гистологические, электронно микроскопические, морфометрические и статистические методы исследования. Результаты. После интрамиокардиального введения БМА в ишемизированный миокард площадь рубца была меньше в 2,74 раза по сравнению с контрольной группой, где биоматериал не применялся. В опытной группе на фоне некротически поврежденных кардиомиоцитов формировался регенерат, состоящий из рыхлой волокнистой соединительной ткани. Отмечалась высокая степень васкуляризации на протяжении всего эксперимента, частицы БМА фагоцитировались макрофагами, продукты биодеградации БМА способствовали ингибированию фиброгенеза, а БМА оказывал кардиопротекторный эффект и индуцировал клеточную регенерацию в ишемически поврежденном миокарде. В периинфарктной зоне зафиксированы разрозненные малодифференцированные клетки с признаками кардиомиогенной дифференциации, наряду с юными кардиомиоцитами выявлены в большом количестве клетки Аничкова. Заключение. Интрамиокардиальное введение БМА уменьшает площадь рубца при инфаркте. Продукты биодеградации БМА снижают скорость рубцевания, оказывают кардиопротективный эффект и индуцируют клеточную регенерацию ишемически поврежденного миокарда. Introduction. Modern technologies for recovery of injured myocardium after a heart attack are not sufficiently effective and do not meet requirements for full rehabilitation of patients. Allogeneic biomaterial is a stimulator of soft tissue regeneration and has been used in various fields of medicine, including surgery, traumatology, ophthalmology, etc. The aim of this work was to study stimulation of regeneration with a biomaterial Alloplant (BMA) in the myocardium injured by experimental ischemia. Methods. This experimental study was conducted on 100 Wistar male rats weighing 0.18-0.25 kg. All animals underwent coronary occlusion by ligation of the coronary artery. In the experimental group, a saline suspension of allogeneic biomaterial (12 mg) was injected intramyocardially into the territory of the stenosed artery simultaneously with the artery ligation. In the control group of animals, a physiological solution was administered. Histological methods used in the study included light-optical microscopy for hematoxylin and eosin staining, Mallory staining, electron microscopy, and morphometry. Hearts were taken for the study at 3, 7, 14, 30, and 45 days. Results. After the BMA intramyocardial injection into the ischemic myocardium, the scar area was 68.5% smaller than in the control group, where the biomaterial was not used. In the experimental myocardium with necrotic cardiomyocytes, an area of regeneration formed, which consisted of loose fibrous connective tissue. Extensive vascularization was observed throughout the experiment. BMA particles were phagocytized by macrophages, and products of BMA biodegradation facilitated fibrogenesis. BMA exerted a cardioprotective effect and induced cell regeneration in the ischemic myocardium. In the periinfarction zone, isolated low-differentiated cells with signs of cardiomyogenic differentiation were observed. Along with young cardiomyocytes, a large amount of Anichkov cells was found. Conclusion. Intramyocardial administration of BMA reduced the scar size in myocardial infarction. Products of BMA biodegradation slowed the scar formation, exerted a cardioprotective effect, and induced cell regeneration in the myocardium injured by ischemia.

Scar smoothing out, angiogenesis stimulation and cardiomyogenesis in myocardial infarction still remain pressing issues despite the variety of existing methods. One of the ways to correct them is intramyocardial implantation of an alloplant biomaterial (ABM) suspension. ABM serves as an inhibitor of fibroneogenesis in various tissues with chronic inflammatory processes. No studies have been carried out with regards to acute myocardial infarction. Objective: to assess the dynamics of the number of bFGF-1 + cells and CD68 macrophages, the degree of angiogenesis amidst the use of ABM in the formation of postinfarction scar in the experiment. Materials and methods. Experimental studies were performed on 100 male Wistar rats weighing 0.18–0.25 kg. Coronary artery ligation was performed on all animals. In the experimental group, the ABM suspension (12 mg) was injected intramyocardially. We used histological, electron microscopic, immunohistochemical (CD68, bFGF-1), morphometric and statistical research methods. Hearts were procured at day 3, 7, 14, 30, and 45. Results. The use of an allogeneic biomaterial immediately after coronary artery stenosis could reduce the area of cicatricial myocardial degeneration by two fold by accelerating inflammatory response and the onset of early proliferative phase. In the reactive zone after ABM implantation, macrophage myocardial infiltration significantly decreased in comparison to the control group. The use of ABM ensures significant predominance of bFGF-1+ cells in the initial period of inflammation (3–14 days). Subsequently (14–45 days), inflammatory cytokine expression became several times less, which corresponded to biodegradation and resorption of the biomaterial. In the control group, during the acute phase of inflammation (3–14 days), bFGF-1+ cells were low in number. Subsequently (14–45 days), cytokine expression increased significantly, causing rapid accumulation of collagen fibers and scarring. In myocardial regeneration after a heart attack in the experiment, ABM stimulated angiogenesis, whose level was three times higher than in the control group. It was noted that ABM serves as a regulator of the neofibrillogenesis-fibroclasia balance in tissue. Conclusion. Macrophage migration inhibition and suppression of pro-inflammatory orientation of macrophages should be indicated as one of the directions of therapeutic correction strategy for ischemic myocardial injuries. Alloplant biomaterial used in the acute phase of myocardial inflammation can serve as such alternative.

Macrophages as the effector cells play a key role in initiating the inflammatory process and predetermine the manifestation of the postinfarction cardiosclerosis. The population of these cells is heterogenous and is mainly represented by M1 and M2 phenotypes. Alloplant biomaterial (ABM) is resorbed by the macrophages which became the regulators of the cellular interaction in tissues. The aim of the investigation was to reveal the peculiarities of the postinfarction healing of the myocardium following the ABM insertion and to assess the population change in the dynamics of macrophages and c-kit+ cells. Materials and methods. The experimental investigations were carried out on 100 male Wistar’s rats weighing 0.18–0.25 kg. All the animals had coronary occlusion by way of ligating the arteries. In the experimental group, the ABM (12 mg) suspension was intramyocardially administered simultaneously with the vessel stricture formation. The harvesting of hearts was carried out at 3, 7, 14, 30, 45 days. Results. In the experimental group the course of the inflammatory process was characterized by the onset of the early proliferative stage, whereas in the control group colliquative necrosis was developing. It was caused by different degrees of the macrophage reaction expression. The number of CD68+ cells in the rat reactive zone of the control group was bigger than in the experimental one. In the experimental group the ABM-induced macrophages of mesenchyme origin were revealed and с-kit+ cells were considerably more in number than in the control one. After 45 days, the scar area index in the experimental group was significantly less than in the control group. Conclusion. ABM had a histoprotective effect under the conditions of the acute myocardial ischemia due to the inhibition of macrophage migration and induction of cellular cardiomyogenesis.

Aim. To identify the features of myocardial remodeling after implantation of a dispersed allogenic biomaterial (DAB) in the subacute stage of experimental infarction and determine the role of decay products in the formation of regenerate.Material and methods. Male rats in the control group (n=30) 5 days after coronary occlusion received physiological saline (intramyocardially), in the experimental group (n=30) — 3 mg of DAB. Myocardial tissues were examined after 7 14, 50 days from the beginning of the experiment using histological, immunohistochemical and statistical methods.Results. In the experimental group, the cicatrix area was reduced by 1,66 times. The products of DAB biodegradation were collagen, sulfated (dermatan- and keratan sulfate), as well as non-sulfated (hyaluronic acid) glycosaminoglycans. Used DAB served as a promoter of the early proliferative phase of inflammation and had an anti-inflammatory effect. DAB particles were a chemoattractant of stem and progenitor myocardial cells in the subacute stage of myocardial infarction. It contributed to their differentiation and integration into the tissue.Conclusion. The products of biodegradation of allogeneic biomaterial, administered intramyocardially in the subacute stage of infarction, contribute to the formation of muscular-connective tissue regenerate and effectively prevent its cicatricial devolution.

Aim. To assess the expression of ММР-9 and TIMP-2 after intramyocardial injection of the allogenic biomaterial (AB) into ischemized myocardium in rodents.Material and methods.One hundred Wistar rodents were used, who underwent ligation of coronary artery. The experiment group (n=50), together with coronary occlusion, received the suspension of AB 12 mg. General histology was applied: hematoxyline and eosine, by Mallori; immune histochemistry (ММP-9, TIMP-2, CD68); morphometry: index of the scar area (ISA), amount of ММP-9, TIMP-2, CD68-positive cells.Results.It was shown that in the experimental group, after implantation of AB there were lesser than in controls values of ISA (2,74 times, p<0,05 . <0,0001), numbers of MMP9+ cells (p<0,001) and macrophages CD68+ (p<0,05), as the manifestation of proteolysis and area of lesion. On the contrary, amount of Timp-2+ cells in the experimental group was higher than in the control (p<0,0004).Conclusion.Resorption of allogenic material (Alloplant) products does influence the balance of metalloproteinases and their inhibitors in the myocardium after ischemic lesion. This effect might influence the processes of myocardial remodelling and scar formation.

We studied the effect of different concentrations of the allogeneic biotransplant on myocardial recovery. In Wistar rats, coronary artery was ligated and intramyocardial injection of 12 or 24 mg Alloplant biomaterial suspension was performed. Histological analysis was conducted on paraffin sections stained by Mallory. The index of the scar area was measured on preparations of transverse sections of the hearts. Allogeneic biomaterial produced cardioprotective and regenerative effect in the myocardium damaged by ischemia. After administration of Alloplant in a dose of 12 mg, the index of scar area decreased by 2.74 times; after doubling the Alloplant dose (24 mg), the index of scar area decreased by 26 times.

Summary. Local progenitor cells are present in all tissues, including the myocardium. Stimulation of these cells can significantly improve the efficiency of the regeneration process. Biodegradation products of Alloplant biomaterials (ABM) are an inhibitor of fibrosis and serve as an inducer of stem cells in various body tissues including smooth and skeletal muscles, as well as connective tissue.Aim. To determine the cell regenerative potential of progenitorial c-kit and committed GATA-4 cells in ischemic myocardium on the background of ABM application in chronic experiment.Materials and methods. 100 Wistar rats weighing 180-200g were used, that underwent ligation of coronary artery. In the experimental group (n = 50), 600 μl of a physiological solution containing 12 mg of ABM was administered simultaneously with coronary occlusion. General histological studies (hematoxylin and eosin staining, and staining as per Mallory), immunohistochemical (c-kit, GATA-4) and morphometric studies (determination of the index of the scar's area, counting of stained cells) were carried out. The statistical processing of the results was carried out using the rank methods - single factor dispersion analysis as per Kruskalou-Wallace and the comparison of uncorrelated data by the Mann-Whitney method.Results. When using ABM during coronary occlusion, myocardium was characterized by a statistically significant (p <0.0001) prevalence of c-kit+ / GATA-4+ cells numbers throughout the follow-up period (3-45 days). These cells were localized mainly in the granulation tissue of the peri-cicatricial area, bordering on the preserved cardiac muscle tissue. In the peri-cicatricial area, cardiac cells had a different degree of maturity - from poorly differentiated forms (without clear ultrastructural signs of cardiomyogenic orientation) to young cardiomyocytes. In the presence of ABM, coronary occlusion was characterized by statistically significantly smaller values of the index of the scar's area (p <0,05 ÷ <0,0001).Conclusion. ABM degradation products are chemoattractants for c-kit cells and promote their differentiation into cardiomyogenic GATA-4 cells, which contributes to more successful myocardium repair.

Abstract. We have performed immunological studies in forty-five patients who underwent lamellar keratoplasty by means of Alloplant biomaterial. Serum antibodies to eye tissue-specific antigens (bovine corneal protein (BCP)-54, lens alpha-crystallin, S-antigen) were tested, and IgG, A, M in the lacrimal fluid were also determined, to detect cellular sensitization against same antigens, and to evaluate the levels of local non-specific inflammation. Percentage of cells activated for BCP-54 was found to be significantly decreased after surgery (p < 0.04), followed by increase to initial levels (p > 0.20) at later terms. Percentage of cells activated for lens α-crystallin was shown to be decreased upon further observation (p < 0.04). A post-surgical increase in IgG levels was revealed in lacrimal fluid (p < 0.02), followed by a decrease later on (p < 0.002). Thus, application of Alloplant biomaterial for lamellar keratoplasty does not cause any sufficient tension of specific humoral immunity against ocular tissues, as evidenced by lack of statistically significant changes of humoral and cellular immunity during post-surgical observations.