Цель работы: изучить динамику состояния клеток крови и костного мозга при прогрессирующем развитии сахарного диабета 2 типа (СД2) в зависимости от степени тяжести нарушений окислительно-восстановительных процессов (ОВП) в тканях организма. Методика. Использована генетическая модель СД2 у мутантных мышей – db/db (опытная группа n=30). Контролем слу жили здоровые мыши той же линии – db/+m (n=10) и линии B10 (n=5). В течение 6–6,5 мес контролировали: динамику кли нических показателей (глюкоза крови, гликозилированный гемоглобин, масса тела) и состояние ОВП в тканях организма по уровню активности НАДН, ФАД и показателю окислительного метаболизма (ПОМ) с помощью аппарата «Лазма-СТ». В течение того же срока исследовали состояние клеток крови (эритроциты, лейкоциты, тромбоциты) и костного мозга. Ста тистическую обработку результатов проводили с предварительным использованием теста Шапиро–Уилкса; достоверность различий с контролем оценивали с помощью параметрического t – критерия Стьюдента, при р<0,05. Результаты. В развитии СД2 выявлено 3 стадии прогрессирующего нарушения метаболизма и ОВП: I – стадия адаптации (1,0-2,0 мес); II – стадия прогрессирующей дисадаптации (2,5-4,5 мес); III – стадия декомпенсации (5,0-6,5 мес). Установлено, что уже в I стадии у мышей db/db снижалось содержание эритроцитов, Hb г/л и лейкоцитов. Во II и особенно в III стадиях происходило повышение тромбоцитов, нейтрофилов, моноцитов, эозинофилов, снижение лимфоцитов. В костном мозге у мышей db/db уже в I, но особенно в III стадии определялось снижение доли живых и повышение количества поврежден ных клеток, преимущественно за счёт апонекротических клеток. Заключение. По мере прогрессирования СД2 и выраженного снижения эффективности ОВП, особенно на III стадии – в организме тормозятся процессы кроветворения и усиливаются нарушения в соотношении клеточных популяций нейтро филы/лимфоциты, что свидетельствует о развитии тяжёлой гипоксии, активации системной воспалительной реакции и торможении репаративных процессов, создающих условия для развития опасных осложнений. Aim: to study the dynamics of changes in the blood and bone marrow cells during the progression of type 2 diabetes mellitus (DM2) depending on the severity of redox (RO) disorders in tissues. Methods. A genetic model of DM2 in mutant db/db mice (experimental group, n=30) was used. The control group consisted of the same mouse strain, db/+m (n=10), and the B10 strain (n=5). Time-related changes in clinical variables (blood glucose, HbA1c, body weight) and the tissue RO status were monitored for 6-6.5 months. The RO status was evaluated by the NADH concentration, FAD activ ity, and the indicator of oxidative metabolism (IOM) using a Lasma-ST apparatus. During the same period, the condition of blood cells (erythrocytes, leukocytes, platelets) and bone marrow cells was examined. Statistical analysis was performed with a preliminary Shap iro-Wilks normality test followed by the parametric Student’s t-test. Differences from the control were considered significant at p<0.05. Results. During the development of DM2, three stages of progressive metabolic and RO disorders were identified: I: stage of adap tation (1.0-2.0 months); II, stage of progressive maladaptation (2.5-4.5 months); III, stage of decompensation (5.0-6.5 months). In db/db mice already at stage I, erythrocytes, Hb and leukocytes were decreased. At stages II and especially III, platelets, neutrophils, monocytes, and eosinophils were increased whereas lymphocytes were reduced. In the bone marrow of db/db mice, already in stage I, but even more in stage III, the proportion of living cells was decreased and the proportion of damaged cells was increased, primarily due to the contribution of aponecrotic cells. Conclusion. With the progression of DM2 and a pronounced decrease in the effectiveness of RO processes, especially at stage III, the hematopoietic processes were inhibited and the disorders in the neutrophil/lymphocyte cell population ratio intensified. This indicates the development of severe hypoxia, activation of a systemic inflammatory response, and inhibition of reparative processes that create conditions for development of dangerous complications.

Objective: to examine how the severity of tissue metabolic disorders affects the dynamics of the state of blood cells and bone marrow (BM) cells in patients with progressive diabetes mellitus (DM).Materials and methods. The genetic model of type 2 diabetes (T2DM) in db/db mutant mice (experimental group, n = 30) was used. Healthy mice of the same line – db/+m (n = 10) and line B10 (n = 5) served as control. The dynamics of laboratory and clinical parameters (blood glucose, glycosylated hemoglobin, body weight) and oxidative metabolism indicators in tissues were monitored FOR 6–6.5 months using Lasma-ST device. The state of blood cells (red blood cells, white blood cells, platelets) and BM cells were examined during the same period. Statistical processing of the results was done with preliminary use of the Shapiro–Wilk test; the significance of differences with the control was assessed using the parametric Student’s t test, at p < 0.05.Results. In the development of T2DM, 3 stages of progressive metabolic disorders were identified: I – adaptation stage (1–2 months); II – progressive maladaptation stage (2.5–4.5 months); III – decompensation stage (from 5.0–6.5 months to death). It was found that in T2DM mice, blood content of red blood cells, Hb and leukocytes was reduced already in stages I–III; but in stage II and especially in stage III, there was increased platelet count and percentage of neutrophils, monocytes, eosinophils with a decrease in lymphocytes. A high percentage of live cells is preserved in the BM in stages I, II and early periods of stage III; in late periods of stage III, live cell percentages are frequently found to be low; in all periods of stage III, the total cell content in the BM is clearly reduced.Conclusion. Hematopoietic processes are inhibited in the BM as T2DM progresses. Individual assessment of the state of BM and its cells at the progressive stages of T2DM may be useful for prognostic purposes.

The progressive development of diabetes mellitus (DM2) in db/db mice is associated with three stages of impaired carbohydrate metabolism and redox processes. In db/db mice, the content of erythrocytes, hemoglobin, and leukocytes was found to decrease already in stage I of DM2. In the II and, in particular, III stages, an increase in platelets, percentage of neutrophils, monocytes, and eosinophils was observed, along with a decrease in lymphocytes. In the bone marrow of db/db mice, a decrease in the proportion of living cells and an increase in the number of damaged cells were determined already in stage I, but especially in stage III, mainly due to aponecrotic cells. Therefore, along with the progression of DM2 and a marked decrease in the effectiveness of redox processes, particularly in stage III, hematopoiesis processes are inhibited and disturbances in the ratio of neutrophil/lymphocyte cell populations increase. This indicates the development of severe hypoxia, activation of a systemic inflammatory reaction, and inhibition of reparative processes that create conditions for the development of dangerous complications.

Введение. Согласно данным литературы Аппарат лазерной диагностики «ЛАЗМА СТ» не был ранее использован для доклинических исследований тканевых нарушений на мелких лабораторных животных. Цель исследования – изучение возможности использования аппарата лазерной диагностики «ЛАЗМА СТ» на мелких лабораторных животных – мутантных мышах линии С57BL/KsJYLeprdb/+ (db/db) – в качестве новой тест системы, для оценки тканевых изменений при сахарном диабете 2 типа (СД 2). Методика. Патологические изменения у мышей с СД изучали на генетической модели СД 2 у мутантных мышей С57BL/KsJYLeprdb/+ (db/db) (n=40); для контроля использовали группу фенотипически здоровых гетерозиготных мышей той же линии (db/+m) (n=16). Общее количество животных составляло 56 голов. Исследование проводили на аппарате лазерной диагностики «ЛАЗМА СТ», который адаптирован приспособлением, ограничивающим подвижность мышей во время измерений. Аппарат «ЛАЗМА СТ» позволяет осуществлять одновременный неинвазивный контроль состояния микроциркуляци крови и лимфы, а также определять уровень активности митохондриальных окислительных коферментов в тканях. Дополнительно измеряли уровень глюкозы в крови фотометрическим методом на приборе Accu-Chek (Швейцария). Результаты. Использование аппарата ЛАЗМА СТ позволило в реальном времени неинвазивно изучить в динамике нарушения микроциркуляции (крови и лимфы) и окислительного метаболизма (НАДН и ФАД) при СД 2, сопоставить их с нарушениями глюкозы в крови, а также прогнозировать тяжесть выявленных нарушений в условиях компенсации, субкомпенсации и декомпенсации. Аппарат ЛАЗМА СТ позволил одновременно выявить начавшиеся нарушения липидного и белкового обмена (пигменты-липофусцин и порфирин), которые свидетельствуют о тяжести прогноза заболевания. Выводы. Аппарат ЛАЗМА СТ, адаптированный для применения у мышей с генетической моделью СД 2 типа, представляет собой новую неинвазивную, информативную и безопасную тест-систему, позволяющую в динамике контролировать и прогнозировать тяжесть нарушений микроциркуляции и тканевых окислительно-восстановительных коферментов – НАДН и – ФАД. The aim of this study was to evaluate a possibility of using the LAZMA ST laser diagnostic apparatus in small laboratory animals, C57BL/KsJYLeprdb/+ (db/db) mutant mice, as a new test system for assessing changes in tissues in type 2 diabetes mellitus (DM2). Methods. Pathological changes were studied on a genetic model of DM2 in C57BL/KsJYLeprdb/+ (db/db) mutant mice (n=40). Phenotypically healthy heterozygous mice of the same strain (db/+m) (n=16) were used as a control group. The study was performed with a LAZMA ST laser diagnostic apparatus that was adapted for mice with a device limiting their mobility during measurements. LAZMA ST allows simultaneous noninvasive monitoring of blood and lymph microcirculation along with measurements of tissue activity of mitochondrial oxidative coenzymes. Additionally, blood glucose was measured photometrically with an Accu-Chek (Switzerland) glucometer. Results. The use of LAZMA ST allowed noninvasive, real-time evaluation of disorders in blood and lymph microcirculation and oxidative metabolism (NADH and FAD) in DM2, comparing them with glycemic disorders, and also predicting severity of these disorders in the conditions of DM2 compensation, subcompensation and decompensation. The LAZMA ST device provided simultaneous detection of the onset of lipid and protein metabolic disorders (lipofuscin and porphyrin pigments) to predict severity of the disease. Conclusion. The LAZMA ST device adapted for using in a murine genetic model of DM2 is a new, noninvasive, informative, and safe test system that allows to monitor and predict the dynamics of severity of disorders in microcirculation and tissue redox coenzymes, NADH and FAD.

С целью выявления критериев прогнозирования тяжести повреждения печени и эффективности корригирующей терапии в обзоре обсуждаются особенности прогрессирования дисфункции иммунитета при хронических фиброзирующих заболеваниях печени. Несмотря на различия в этиопатогенезе хронических заболеваний печени, развитие фиброза и цирроза печени происходит на фоне однотипно прогрессирующих проявлений дисфункции компонентов врождённого и адаптивного иммунитета, а также изменений костного мозга – центрального органа иммуногенеза. Наблюдаемые сдвиги формируются как следствие хронического воздействия на организм интоксикации, транслокации бактерий из кишечника, нарушенного метаболизма и прогрессирующего системного воспаления. Уже на раннем этапе активации процессов фиброзирования печени клетки врождённого и адаптивного иммунитета становятся длительно гиперактивированными, появляются субпопуляции с профиброгенными иммуносупрессирующими свойствами (стадия субкомпенсации). По мере прогрессирования деструктивных процессов в печени (переход фиброза в цирроз) в состоянии иммунных клеток появляются признаки истощения функции, выраженного цитокинового дисбаланса и стойкой иммуносупрессии (стадия декомпенсации или «иммунного паралича»). Иммуносупрессия при этом становится фактором повышенной восприимчивости организма к бактериальным инфекциям и септическим осложнениям, а также фактором глубокого торможения регуляции восстановительных процессов (второй важнейшей функции иммунитета) и развития необратимости повреждения печени. Стадийность развития иммунного дисбаланса при прогрессировании фиброза в цирроз может быть выявлена с помощью маркеров врожденного и адаптивного иммунитета, а также по содержанию в крови CD34+ клеток костного мозга. Обсуждается значение выявления стадий иммунного дисбаланса для прогнозирования тяжести (обратимости) повреждения печени и эффективности применения корригирующей терапии. This review discusses features of progressive immune dysfunction in chronic fibrosing liver diseases to identify criteria for predicting the severity of liver (L) damage and the effectiveness of corrective therapy. Despite differences in the etiopathogenesis of chronic L disease, the development of L fibrosis and cirrhosis is associated with similarly progressing manifestations of dysfunction of innate and adaptive immunity components, as well as changes in the bone marrow, the central organ of immunogenesis. These changes result from chronic intoxication, bacterial translocation from the gut, impaired metabolism, and progressive systemic inflammation. Already at the early stage of L fibrosis activation, innate and adaptive immune cells become chronically hyperactivated, and subpopulations with profibrogenic immunosuppressive properties emerge (subcompensation stage). With the progression of hepatic destructive processes (evolution of fibrosis to cirrhosis), the immune cells display signs of functional exhaustion, pronounced cytokine imbalance, and persistent immunosuppression (decompensation stage or “immune paralysis”). In this case, immunosuppression becomes a factor of increased susceptibility of the body to bacterial infections and septic complications, as well as a factor of deep inhibition of the regenerative process (the second most important immunity function) and the development of irreversible L damage. The staging of the immune imbalance during the progression of L fibrosis to cirrhosis can be detected with markers of innate and adaptive immunity and by the blood content of CD34+ bone marrow cells. The authors discussed the importance of identifying the stages of immune imbalance for predicting the severity (reversibility) of L damage and the effectiveness of corrective therapy.

Creation of a bioartificial pancreas, including a cell-engineered construct (CEC) formed from pancreatic islets (islets of Langerhans) and a biocompatible matrix mimicking the native microenvironment of pancreatic tissue, is one of the approaches to the treatment of type 1 diabetes mellitus (T1D). Objective: to conduct preliminary in vivo studies of the functional efficacy of intraperitoneal injection of a cell-engineered pancreatic endocrine construct and a suspension of rat pancreatic islets in an experimental T1D model. Materials and methods. Tissue-specific scaffold was obtained by decellularization of human pancreatic fragments. The viability and functional activity of rat islets isolated with collagenase were determined. Experimental T1D was modeled by intraperitoneal injection of low-dose streptozotocin and incomplete Freund's adjuvant into rats. The rats were intraperitoneally injected twice with pancreatic CEC (n = 2) or islet suspension (n = 1). Glucose levels in the blood and urine of the rats were assessed. Histological examination of organs (pancreas and kidneys) of the experimental animals was carried out. Results. After the first injection, blood glucose levels gradually decreased in all animals by more than 47% of the initial values; by follow-up day 24, the glucose level rose to the initial hyperglycemic values. After repeated administration, a 63.4% decrease in glycemic level was observed in the rats with pancreatic CEC and a 47.5% decrease in the one with islet suspension. At week 5 of the experiment, blood glucose levels gradually increased in all animals. At the same time, the glycemic index of the rat with injected pancreatic CEC was 62% lower than the glycemic index of the rat with injected islets. Conclusion. Allogeneic pancreatic islets in pancreatic CEC increase the duration of stable glycemic level in T1D rats.

Objective: to study the effectiveness of correcting the morphofunctional characteristics of the liver in an experimental model of chronic liver disease (CLD), using implanted cell-engineered constructs (CECs).Materials and methods. Experiments were carried out on male Wistar rats (n = 80) aged 6–8 months with an initial weight of 230–250 g. CLD was modeled by inoculating the rats with 60% CCl4 oil solution for 42 days based on a modified scheme. Microgel based on recombinant spidroin rS1/9 was used as a matrix for CECs fabrication. Allogeneic liver cells (LCs) and multipotent bone marrow-derived mesenchymal stem cells (BM-MSCs) from a healthy donor were used as the cellular component of the CECs. The effectiveness of the corrective effect of the implanted CECs was assessed in an experimental CLD model (n = 60) in two groups of rats: Group 1 (control, n = 20, 1 mL of saline solution was injected into the damaged liver parenchyma) and Group 2 (experimental, n = 40, CECs containing allogenic LCs and BM-MSCs in a 5 : 1 ratio in a volume of 1 mL were implanted into the damaged liver parenchyma). For long-term monitoring of the CEC state, the CECs were labeled by additional inclusion in Cytodex-3. The effectiveness of the regulatory effect of CECs on regenerative processes in the liver was evaluated using biochemical, morphological and morphometric techniques, as well as by flow cytometry at 90 days after implantation.Results. In the control group, the mortality rate in CLD was 25%. There was no death in the experimental group with CLD after CEC implantation. The CECs were found to have a corrective effect on the biochemical and morphological parameters of the liver in CLD during 90 days of follow-up, with concomitant preservation of structural cellular homeostasis in the implanted CECs. Conclusion. Implantation of CECs in the liver facilitates effective correction of CLD by activating regenerative processes in the damaged liver, which is due to long-term preservation of structural cellular homeostasis in the CECs.

Objective: to compare the efficiency of regenerative processes in the liver using apoptotic bone marrow-derived mononuclear cells (BMMCs) and intact BMMCs from healthy animals on an extended liver resection (ELR) model. Materials and methods. Male Wistar rats (n = 77) with an ELR model (70–75%) were divided into 3 groups: group 1 (control with a single intraperitoneal injection of saline), group 2 (single intraperitoneal injection of unsorted intact BMMCs at a dose of 30–35 × 106, and group 3 (single intraperitoneal injection of apoptotic BMMCs at the same dose). Restoration of biochemical parameters of liver function and mass, as well as the emerging microstructural changes in hepatocytes in histological preparations, were monitored by assessing hepatocyte mitotic activity (MA) during the first 7–10 days after ELR. Results. It was found that in groups 2 and 3, as compared with group 1, there was no death after ELR modeling, and that the biochemical parameters of liver function normalized more rapidly (at days 10–14). Hepatocyte MA in group 3 sharply increased as early as on day 1, and mitotic index (MI) averaged 14‰, reaching 20.9‰ in some experiments; MI in the control group remained at the baseline by this time, while in group 2, MI was only 3.2‰. In group 3, liver mass recovered more rapidly after ELR to baseline values already at days 8–10, whereas the recovery was at day 12–14 and day 17–20 in group 2 and group 1, respectively. It was suggested that the more pronounced increase in the efficiency of regenerative processes in the liver after ELR in group 3 after using apoptotic BMMCs was due to the release from these cells of a large spectrum of formed paracrine factors, including various classes of RNA molecules involved in the regeneration process. Conclusion. Apoptotic BMMNCs have a more effective adaptive and regulatory potential than intact BMMCs because reorganizations are rapidly formed in the damaged liver cells, providing an early and more powerful activation of the targeted regenerative program.

Cell death represents the most critical pathologic entity in liver disease, which dictates pathologic consequences such as inflammation, fibrosis, and cell transformation. We analyzed the conclusions of studies on the involvement of different types of programmed cell death (PCD) in the pathogenesis of liver diseases. Three main forms of PCD (autophagy, apoptosis, necrosis) and five additional, still insufficiently studied PCD – necroptosis, ferroptosis, pyroptosis, partanatosis and entosis – observed in the liver in various acute and chronic diseases are considered. The involvement of several PCD at once in the development of any one pathology and one type of PCD in different pathologies was established. This indicates the existence of cross-regulation of metabolism in the liver cells with different levels of damage in the formation of the main dominant type of PCD. Available results indicate the possibility of attenuation (correction) of functional and morphological manifestations of PCD in the organ by controlled blocking of effector-mediated PCD pathways, as well as targeted induction of autophagy, anti-apoptotic and anti-necrotic mechanisms in liver cells.

Objective : to study the peculiarities of the induction effect of total RNA (tRNA) from xenogenic bone marrow cells (BMCs) on regeneration processes in the recipient's native liver with extensive liver resection using an adoptive transfer model. Materials and methods . The study was carried out on an adoptive transfer model using male Wistar rats (n = 20) and guinea pigs (n = 17). The donors were rats (n = 10). 12 hours after extensive liver resection (70-75%), tRNA was isolated from BMCs and injected into intact (non-operated) recipients intraperitoneally at a dose of 30 μg/100 g of weight. The induction effect of the tRNA on operated rats was studied in 3 groups of recipients: Group 1 (control, n = 5) - administration of saline to guinea pigs; Group 2 (control, n = 10) - administration of tRNA from a donor rat to a recipient rat (allogeneic transfer); Group 3 (experiment, n = 12) - administration of tRNA from a donor rat to a recipient guinea pig (xenogeneic transfer). In histological preparations of recipient livers, after 48, 72 hours and 7 days, we studied the mitotic activity of hepatocytes and the features of the microscopic picture of the liver. The significance of differences in the compared groups was assessed using the parametric Student's t-test. Results. The ability of BMC tRNA to tissue-specifically activate regenerative and immune responses in the liver after extensive resection was found to depend on the donor and recipient species identity. Introduction of allogeneic donor tRNA in the recipient's liver resulted in predominant enhancement in hepatocyte mitotic activity (p < 0.05). The use of xenogeneic donor tRNA leads to enhanced activity of only immuno-inflammatory reactions in the recipient's liver, such as sinusoidal cell activation, lymphocytic infiltration into sinusoids, and portal tract infiltration by inflammatory cells. Conclusion. To induce regenerative processes in the liver, tRNA obtained from allogeneic BMCs should be used.

Objective: using an adoptive transfer model to study the cellular mechanisms involved in the formation of the initial stage of liver regeneration during intraperitoneal injection of a healthy recipient with apoptotic bone marrowderived mononuclear cells (BM-MNCs) from a donor after extended liver resection.Materials and methods. Male Wistar rats (n = 40) were used to create a model of adoptive transfer of apoptotic BM-MNCs (a-BM-MNCs) taken from the donor after extended liver resection to a healthy recipient. During the experiments, the animals were divided into five groups. Four experimental groups with intraperitoneal injection of the same doses to the recipient: freshly isolated BM-MNCs (group 1); BM-MNCs subjected to apoptosis for 48 hours by storage at t = 4–6 °C in phosphate-buffered saline (PBS) (group 2) or in a Custodiol HTK solution (group 3). In group 4, the animals were injected with PBS after storing BM-MNCs in it. The control animals were animals injected with saline (group 5). For selection of effective modes of apoptosis induction, BM-MNCs stained with 7AAD after incubation in solutions were analyzed by flow cytometry. Targeted transfer of regenerative signals to the recipient was assessed by the mitotic activity of hepatocytes in the liver and tubular epithelium in the kidneys, as well as by the intensity of microstructural changes in the liver 24, 48 and 72 hours after injection of the studied material.Results. BMC incubation in PBS and HTK for 48 hours at t = 4–6 °C provides the most effective accumulation of a-BM-MNCs in early apoptosis. It was shown that a-BM-MNCs retain the ability to target-focused transmission of regulatory signals to the liver supported by autophagy process during adoptive transfer. It was established that a-BM-MNCs (groups 2 and 3) in comparison to native BM-MNCs (group 1) at adoptive transfer increased the regenerative potential of the liver due to pronounced increase in the activity of autophagy processes and directed infiltration of immunomodulatory mononuclear cells in the liver.Conclusion. a-BM-MNCs create a stronger basis for development and implementation of a targeted and effective regeneration program by enhancing autophagy processes and immunomodulatory effect on mononuclear cells, which are regenerative signal carriers.

Objective: to study the cellular mechanisms of activation of regenerative processes in the liver when using total RNA (tRNA) of bone marrow cells (BMCs) based on an extended liver resection (ELR) model. Materials and methods. Male Wistar rats (n = 80) with ELR model (70%) were divided into 2 groups: group 1 (control group) had a single saline injection, while group 2 (experimental group) received a single tRNA injection at a 30 μg/100 g dose of animal weight. The biochemical parameters of liver function and weight were monitored over time. Also monitored were microstructural changes in hepatocytes 48 hours after ELR by examining mitotic activity, caspase-9 expression and morphometric parameters. Results. It was found that in group 2, in comparison to group 1, there was faster normalization of biochemical parameters (by 10–14 days), a higher mitotic index of hepatocytes (23.45‰ versus 5.37‰), and initially sharper decrease and then faster recovery of liver mass (by 10–12 days versus 18–20 days). Both groups showed almost total expression of caspase-9, including in mitotically splitting hepatocytes. Group 1 demonstrated decreased values of morphometric parameters of single and binuclear cells, decreased number of binucleated hepatocytes and increased total density of hepatocytes as compared to the intact liver. Intraperitoneal administration of tRNA increased morphometric parameters of mononuclear hepatocytes, did not affect their number, but increased the area of the nuclei of binuclear hepatocytes as compared to the control group. Conclusion. The proven capability of cell-bone marrow total RNA to simultaneously support apoptosis in liver cells after ELR and induce mitotic activity indicates that tRNA can switch activated apoptosis to cell proliferation at the early phase of the regenerative process. This effect may be due to the presence of regulatory RNA molecules in tRNA, including numerous non-coding RNAs.

Objective: to investigate the functional efficiency of a cell-engineered construct (CEC) of the liver based on tissuespecific matrix consisting of decellularized rat liver fragments, allogeneic liver cells and multipotent mesenchymal stromal cells (MSCs) isolated from the bone marrow on an experimental model of chronic liver failure (CLF). Materials and methods . In creating liver CECs, the liver for decellularization and liver cells were obtained from male Wistar rats. MSCs were isolated from rat bone marrow. The functional efficacy of CEC was investigated on an experimental CLF model obtained by priming rats with CCl 4 solution. At different periods after implantation, the outcomes were assessed based on the biochemical parameters of cytolysis. Morphological changes in the liver were analyzed by histochemical methods in the control (administration of saline solution into the liver parenchyma) and experimental (administration of liver CEC into the liver parenchyma) groups. Results . It was shown that implantation of the proposed CEC normalizes blood biochemical parameters and structural disorders of the damaged rat liver faster (by day 30 after introduction of CEC instead of day 180 in the control). The CEC was also shown to have reduced animal mortality from 50 to 0%, which is due to early activation of proliferation of viable liver cells and faster formation of new blood vessels. These effects are down to either stimulation of the internal regenerative potential of the damaged liver during CEC implantation or long-term functioning of the transplanted cells as part of the CEC based on the decellularized liver matrix. Conclusion. The liver CEC, implanted into the liver parenchyma in laboratory animals with a CLF model, has a functional activity.

Aim: to carry out a comparative assessment of the effectiveness of recovery processes in the liver after modeling of chronic fibrotic liver damage (CFLD) at using of bone marrow mononuclear cells (BMMC) and total RNA (tRNA), isolated from BMMC. Materials and methods. In this study it was used 140 rats of Wistar breed. CFLD was modeled on 100 rats, of which 25 died. The surviving 75 rats (CFLD formed by 3 months) were divided into 3 groups: group 1 - control (administration of saline solution); group 2 - single administration of tRNA from BMMC at a dose of 30 mu g/100 g weight; group 3 - single administration of BMMC at a dose of (30-35) x 10(6) cells. The dynamics of recovery processes in the liver was evaluated by the mortality of animals, as well as by the dynamics of the recovery of biochemical parameters (A1AT, MAT, ALP and total protein) and morphological status of the liver in 7 days and in 3,6 and 9 months. The reliability of differences between compared values was evaluated by the t - Student criterion at p < 0.05. Results. The mortality of animals with CFLD in group 1 was 12%, in groups 2 and 3 by 4%; In group 1- A1AT and AsAT were restored to normal values in 2 months, ALP in 3 months, and the total protein remained reduced for more than 4 months. In groups 2 and 3, all hepatic homeostasis indices returned to the values, which were before CFLD modeling, faster than in group 1 (in 2 months), but in group 2 the recovery rate was higher than in group 3. It was shown that the normalization of liver functional parameters in all groups was ahead of the restoration of its histological structure. The activation of liver defibrotic processes in group 2 occurred in 3 months, and in groups 1 and 3 - in 6 months. The restoration of liver histological structures in group 2 occurred in 6 months, and in groups 1 and 3 - after 9 months. Conclusion. BMMC and tRNA from BMMC in biologically effective doses take in part at the activation of liver recovery processes after modeling CFLD, but the regulatory impact of tRNA appears earlier and is more effective.

Aim: to determinate the most effective liver cells and multipotent mesenchymal stromal cells of bone marrow (MMSC BM) ratio into implantable cell engineering constructions (CECs) used for chronic liver failure (CLF) correcting.Materials and methods. For creating liver CECs it was used a biopolymer implant – a composition of a heterogeneous collagen-containing gel (BMCG) (Sphero®GEL trademark) containing viable liver cells and MMSC BM in the following ratios – 1 : 1; 5 : 1 and 10 : 1 respectively. CECs with different ratios of liver cells and MMSC BM were implanted into liver of rats in which chronic liver failure (CLF), was modeled by using CCl4. The effectiveness of the regulatory effects of CECs (with different cell ratios) on regenerative processes in livers were assessed by using biochemical, morphological and morphometric methods at different periods after their implantation.Results. Corrective effect of CECs with different cell composition on biochemical and morphological parameters of livers at chronic liver failure was established. During studying the liver CECs with various cell ratios of liver cells and MMSC BM (1 : 1; 5 : 1 and 10 : 1 respectively), it was found that the most optimal ratio of cells into the CECs is 5 : 1, because at this ratio of cells, there were a more distinct normalization of the morphological and functional liver parameters within 365 days after modeling CLF and maintenance of the structural homeostasis into the CECs. Themselves, which allows predicting their long-term regulatory effect on the liver tissue in CLF and maintaining its normal structural and functional state.Conclusion. The effective correction of chronic liver failure can be carried out by using the implanted liver CECs, in which donor liver cells and MMSC BM where presented in ratios – 1 : 1; 5 : 1 and 10 : 1. But analysis of prolonged correction of liver morphological and functional parameters at CECs using it was allow to recommend the preferences using of CECs with ratio 5 : 1, because prolonged preservation of structural homeostasis into these CECs makes possible to prognosticate their prolonged regulatory action on the liver tissue at CLF, especially for recipients on a waiting list for liver transplantation.

Aim: to conduct a comparative assessment of the effectiveness of liver regeneration occurring after induction of chronic fibrosing liver disease (CFLD) using bone marrow mononuclear cells (BMMCs) and total RNA (tRNA) extracted from BMMCs.Materials and methods. The study involved 140 Wistar rats. CFLD was modeled in 100 rats, of which 25 died. The surviving 75 rats (CFLD formed by the third month) were divided into 3 groups: Group 1 – control (administered with physiological saline); Group 2 – a single injection of tRNA from BMMCs at a dose of 30 μg/100g body weight; Group 3 – a single injection of BMMCs at a dose of (30–35) × 106 cells. The dynamics of regenerative processes in the liver was evaluated based on the animal mortality, dynamics of restoration of biochemical markers (ALAT, ASAT, alkaline phosphatase and total protein) and morphological picture of the liver on the seventh day and after three, six and nine months. The significance of differences in the compared values was determined through Student’s t-test for <0.05.Results. Mortality in Group 1 was 12%, in Groups 2 and 3 – 4%; In Group 1, ALAT and ASAT were restored to normal values after two months, alkaline phosphatase after 3 months, and total protein remained low for over 4 months. In Groups 2 and 3, all hepatic homeostasis markers returned to the values they were before CFLD modeling faster than in Group 1 (after two months). However, in Group 2, the regeneration rate was higher than in Group 3. It was revealed that normalization of functional liver parameters in all groups were ahead of restoration of the histological structure of the liver. Liver defibrotic processes in Group 2 were activated after 3 months, and in Groups 1 and 3 – after 6 months. The histological structure of the liver was restored in Group 2 after 6 months, and in Groups 1 and 3 after 9 months.Conclusion. BMMCs and tRNA extracted from them in biologically effective doses trigger liver regeneration in CFLD. However, regulatory effect from the use of tRNA appears earlier and is more effective.

Extensive liver resection (ELR), performed in a number of surgical operations, refers to a critical injury, which necessitates the improvement of methods of therapy of acute post-resection liver failure.Aim:to compare the effectiveness of stimulation of regenerative processes in the liver residue after ELR (60–70%) by intraperitoneal administration of lysed aspirate bone marrow cells (BMCs) and total RNA (tRNA) isolated from BMCs.Materials and methods.This work was performed on 175 rats-male Wistar breed 250–300 g, on 75 of which under the inhalation anesthesia it was reproduced the model of the ELR in three groups of experiments: group 1 – control (administration of isotonic solution after ELR), group 2 – in 3–5 hours after ELR the tRNA from BMCs was intraperitoneally injected at a dose of 30 μg/100 g, group 3 – in 3–5 hours after ELR BMCs was administered intraperitoneally at a dose of 30–35 × 106cells per rat. Comparative studies of the restorative processes in the liver after the ELR in the three groups were carried out by dynamic control of the mitotic activity of hepatocytes in the liver residue, cytolytic enzymes, total bilirubin and total serum protein, as well as the liver residue (mass) weight.Results.The tRNA from BMCs and BMCs in the indicated doses prevent the risk of the development of lethal outcomes, and also contribute to an earlier (by 10–14 days) normalization of the functional indices of hepatic homeostasis. However, the tRNA from BMCs, compared with BMCs, has a stronger stimulating effect on the recovery processes: it promotes earlier intensification of mitotic activity of hepatocytes and provides a higher rate of recovery of liver mass.Conclusion.For the induction of recovery processes in the liver residue after ELR, the preference should be given to the tRNA from BMCs.

Aim : to conduct a comparative assessment of the effectiveness of liver regeneration occurring after induction of chronic fibrosing liver disease (CFLD) using bone marrow mononuclear cells (BMMCs) and total RNA (tRNA) extracted from BMMCs. Materials and methods . The study involved 140 Wistar rats. CFLD was modeled in 100 rats, of which 25 died. The surviving 75 rats (CFLD formed by the third month) were divided into 3 groups: Group 1 – control (administered with physiological saline); Group 2 – a single injection of tRNA from BMMCs at a dose of 30 μg/100g body weight; Group 3 – a single injection of BMMCs at a dose of (30–35) × 106 cells. The dynamics of regenerative processes in the liver was evaluated based on the animal mortality, dynamics of restoration of biochemical markers (ALAT, ASAT, alkaline phosphatase and total protein) and morphological picture of the liver on the seventh day and after three, six and nine months. The significance of differences in the compared values was determined through Student’s t-test for <0.05. Results . Mortality in Group 1 was 12%, in Groups 2 and 3 – 4%; In Group 1, ALAT and ASAT were restored to normal values after two months, alkaline phosphatase after 3 months, and total protein remained low for over 4 months. In Groups 2 and 3, all hepatic homeostasis markers returned to the values they were before CFLD modeling faster than in Group 1 (after two months). However, in Group 2, the regeneration rate was higher than in Group 3. It was revealed that normalization of functional liver parameters in all groups were ahead of restoration of the histological structure of the liver. Liver defibrotic processes in Group 2 were activated after 3 months, and in Groups 1 and 3 – after 6 months. The histological structure of the liver was restored in Group 2 after 6 months, and in Groups 1 and 3 after 9 months. Conclusion . BMMCs and tRNA extracted from them in biologically effective doses trigger liver regeneration in CFLD. However, regulatory effect from the use of tRNA appears earlier and is more effective.

Aim: to determinate the most effective liver cells and multipotent mesenchymal stromal cells of bone marrow (MMSC BM) ratio into implantable cell engineering constructions (CECs) used for chronic liver failure (CLF) correcting. Materials and methods. For creating liver CECs it was used a biopolymer implant – a composition of a heterogeneous collagen-containing gel (BMCG) (Sphero®GEL trademark) containing viable liver cells and MMSC BM in the following ratios – 1 : 1; 5 : 1 and 10 : 1 respectively. CECs with different ratios of liver cells and MMSC BM were implanted into liver of rats in which chronic liver failure (CLF), was modeled by using CCl4. The effectiveness of the regulatory effects of CECs (with different cell ratios) on regenerative processes in livers were assessed by using biochemical, morphological and morphometric methods at different periods after their implantation. Results. Corrective effect of CECs with different cell composition on biochemical and morphological parameters of livers at chronic liver failure was established. During studying the liver CECs with various cell ratios of liver cells and MMSC BM (1 : 1; 5 : 1 and 10 : 1 respectively), it was found that the most optimal ratio of cells into the CECs is 5 : 1, because at this ratio of cells, there were a more distinct normalization of the morphological and functional liver parameters within 365 days after modeling CLF and maintenance of the structural homeostasis into the CECs. Themselves, which allows predicting their long-term regulatory effect on the liver tissue in CLF and maintaining its normal structural and functional state. Conclusion. The effective correction of chronic liver failure can be carried out by using the implanted liver CECs, in which donor liver cells and MMSC BM where presented in ratios – 1 : 1; 5 : 1 and 10 : 1. But analysis of prolonged correction of liver morphological and functional parameters at CECs using it was allow to recommend the preferences using of CECs with ratio 5 : 1, because prolonged preservation of structural homeostasis into these CECs makes possible to prognosticate their prolonged regulatory action on the liver tissue at CLF, especially for recipients on a waiting list for liver transplantation.