Relevance. Creating three-dimensional scaffolds from biodegradable materials and seeding them with stem cells derived from the oral tissues is a promising tool for guided tissue regeneration. Pulp and periodontal stem cells have a high potential for osteogenic differentiation, which biologically determines their use in surgical bone reconstruction. The experiment shows the result of using fibrin glue seeded with pulp and periodontal stem cells on the mandible of laboratory mice. The article presents the results of computed tomography and histological examination. The data provide evidence of the influence of seeded scaffolds on bone remodeling in the area of the defect.Materials and methods. The Local Ethics Committee of the North-Western State Medical University named after I.I. Mechnikov gave permission for the practical part of the research work. The study included 29 white laboratory mice. Molars were extracted and a bone defect was formed. Pulp and periodontal stem cells were obtained and cell-seeded scaffolds were made, then they were introduced into the defect area. The animals were euthanized, maxillofacial CT scan and histology of the defect area were performed 28 days after the molar extraction.Results. The oral cavity of mice was examined, molars were extracted, and teeth were morphologically examined under anesthesia. Scaffolds were synthesized and bone defects were filled. CT scans and histology results were analyzed. The bone volume increased in the main group compared to the control group.Conclusion. The fibrin glue can be used to obtain a material with mechanical characteristics sufficient for a stable shape scaffold. The study proved that the pulp stem cells enclosed in a fibrin glue-based scaffold can maintain the ability to proliferate and osteogenically differentiate. The scaffold based on fibrin glue, which we used, affected the bone remodeling process in the area of jaw defects.

To date, there are quite a large number of reports of the use of stem cells in the treatment of various diseases in humans. However, the authors do not provide credible evidence that the introduced stem cells come directly into the target organ. Without a clear understanding of the location of the introduced stem cells, it is difficult to reliably assess the effect of their use. The article gives its own observation about the possibility of imaging in the human body autologous mesenchymal stem cells introduced through the portal vein of the liver to a patient suffering from cirrhosis. Iron oxide nanoparticles were used as labels. A rather simple question was previously solved: how to create a label of such size that the viability of the stem cell was preserved and it could be visualized in the human body when performing magnetic resonance imaging of abdominal organs. Following in vitro studies, the most appropriate stem cell marking embodiment was chosen to provide both adequate imaging and cell viability. After administration of marked table cells, magnetic resonance imaging of abdominal organs was performed to the patient. The introduced stem cells were visualised only in hepatic tissue. According to clinical and biochemical blood tests - no deviations were detected. There were no complications against the background of this procedure. The patient was discharged for outpatient treatment in satisfactory condition.

Clinical experience of application of autologous multipotent mesenchymal stromal cells in treatment of patient suffering from liver cirrhosis of alcoholic etiology is presented. A special feature is that the hepatocyte precursors were isolated directly from the patients liver tissue. The patient underwent laparoscopic surgery to obtain the largest volume of material and to be able to visually control the tissue sampling with minimal fibrotic changes. After liver tissue sampling, the patient was discharged for outpatient treatment in a satisfactory condition. Subsequently, the patient was re-admitted to the hospital. During repeated hospitalization, multipotent mesenchymal stromal cells in the amount of 20 million were injected into the arterial bed of the liver using x-ray endovascular technique. In the control study, 6 months after treatment, according to the 13C-metacetin test, normalization of liver function, regression of portal hypertension, and an increase in platelet levels were noted. There were no complications during this treatment. Treatment of patients suffering from cirrhosis of the liver is quite a serious and complex task. As a rule, the patient learns about his diagnosis already in the presence of complications, when the liver function is already significantly impaired. The propensity of the population to alcoholism leads to the formation of fibrosis, and subsequently cirrhosis of the liver. The absence of anti-fibrotic drugs contributes to the implementation of research to find alternative methods of treatment for this category of patients. In General, the use of autologous multipotent mesenchymal stromal cells is an effective and promising method, and research in this direction should be continued.

Through clinical observation, we present an assessment of the autologous mesenchymal stem cells effectiveness in treatment of liver cirrhosis of alimentary etiology. In order to determine the localization of the implanted cell structures, the stem cells were previously labeled with iron (II, III) oxide nanoparticles (IONPs). Further MRI visualization helped to detect the cell structures stained with iron oxide nanoparticles in the human body. In 6 months after the cell therapy, the patient underwent clinical and biochemical blood tests, MEGX test, elastography and subjective health assessment test. The tests data analysis revealed the improvement of the values of all examined parameters after the cell treatment. Also in 6 and 12 months after the treatment, a liver biopsy was performed from the area where the implanted stem cells were visualized. In histological examination of liver bioptates obtained from the area of MSC transplantation, the largest number of stained cells was observed in liver micronodes, as well as at the boundaries of micronodes and fibrous septa. A portion of the bioptate obtained in 12 months after transplantation was used to produce primary cell cultures. Before the first re-seeding of the cultures, cell colonies of both fibroblast-like morphology and epithelial were detected in them. Both types of colonies contained the particles. Conducting the cell therapy to a patient with liver cirrhosis of alimentary etiology contributed to improving the laboratory and instrumental examinations indicators. The patient had come through the treatment procedure satisfactorily, no complications were registered.

The increase in the number of patients with diffuse liver diseases, the lack of efficacy from conservative treatments that produce long-lasting and stable outcomes, contribute to the search for alternative treatments. The goal is to assess the effectiveness of using autologous mesenchymal stem cells in treating patients with diffuse liver disease. Materials and methods: the study involved 2 groups of patients of 20 people each. The main group included patients who used autologous mesenchymal stem cells (20-25 million) introduced into the vascular liver channel as therapy. The control group included patients who were treated with L-ornithine-L-aspartate 1 package 3 times a day for 30 days. Benchmarks for evaluation of treatment effectiveness were determined 1, 3 and 6 months after the therapy. Results: in the main group, 3 months after treatment, a normalization of platelet levels was observed, whereas in the control group, platelets did not reach normal values and 6 months after treatment. 1 month after treatment, hepatic enzyme levels were normalized in the main group. The result was maintained 6 months after treatment. In the control group, the level of alanine aminotransferase (ALT), alkaline phosphatase and gamma-glutamyl transpeptidase (GGTP) was above normal throughout the observation; the level of aspartate aminotransferase (AST) normalized to 3 months after treatment and then exceeded normal levels again 6 months from the start of therapy. The functional state of the liver according to the results of the lidocaine test at 6 months after treatment was normalized in 70 % of patients of the main group and in 25 % of patients of the control group. Manifestations of encephalopathy regressed in 65 % of patients in the main group and in 10 % of the control group. After 6 months of treatment, patients with low and reduced level of subjective comfort were absent in the main group, while in the control group such patients were 45 and 35 %, respectively. No complications have been identified against the background of the use of cellular technologies. Findings: the use of cellular technologies is an effective and safe way to treat patients with diffuse liver disease. Continuing research in this direction in the future will help to create algorithms for the use of stem cells in the treatment of patients with diffuse liver diseases.

В клиническом наблюдении представляется оценка эффективности использования аутологичных мезенхимальных стволовых клеток в лечении цирроза печени алиментарной этиологии. Для определения локализации вводимых клеточных структур предварительно проводилось мечение стволовых клеток наночастицами оксида железа (II, III). Клеточные структуры, отмеченные наночастицами оксида железа, были визуализированы в организме человека при помощи МРТ. Через 6 месяцев после проведения клеточной терапии у пациента были оценены показатели клинического и биохимического анализов крови, данные MEGX-теста, показатели эластографии и теста субъективной оценки самочувствия. После проведения клеточной терапии отмечалось улучшение показателей всех вышеперечисленных методов исследования. Также через 6 и 12 месяцев после лечения была выполнена биопсия печени из области, где были визуализированы введенные ранее стволовые клетки. При гистологическом исследовании биоптатов печени, полученных из области трансплантации МСК, наибольшее количество меченых клеток наблюдали в микроузлах печени, а также на границах микроузлов и фиброзных септ. Часть биоптата, полученного через 12 месяцев после трансплантации, использовали для получения первичных культур. До первого пересева культур в них выявляли колонии клеток как фибробластоподобной морфологии, так и эпителиальной. Оба типа колоний содержали частицы. Проведение клеточной терапии пациенту с циррозом печени алиментарной этиологии способствовало улучшению показателей лабораторных и инструментальных методов исследования. Процедура была перенесена пациентом удовлетворительно, каких-либо осложнений выявлено не было.

This study evaluated the feasibility of using uncoated iron (II, III) oxide nanoparticles (IONP) obtained by electric explosion of wire in air for labelling living mesenchymal stromal cells and their subsequent visualization by magnetic resonance imaging (MRI) using 1.5T clinical MRI scanners. The uptake of uncoated IONP by MSC was demonstrated for the wide range of IONP concentration in the cell culture medium. The cells did not change their proliferative activity, viability, and the set of surface markers. IONP obtained by electric explosion of wire in an atmosphere of air had a shape close to spherical. The size of nanoparticles varied from 14 to 136 nm according to dynamic lateral light scattering, laser diffraction, and transmission electron microscopy. Particles up to 136 nm comprised 75%, and particles less than 36 nm --- 10% of the IONP powder. A wide range of particle sizes made it possible to select MRI parameters suitable for labelled cells detection in animal tissues both in the T2 mode and in the T1 relaxation mode.

Abstract. The effectiveness of the influence of autologous mesenchymal stem cells on the function of liver tissue in liver cirrhosis of alimentary etiology is considered, as well as the possibility of visualization of the introduced cell structures with subsequent cytological confirmation. To be able to track autologous mesenchymal stem cells introduced in the patients body, they were labeled with iron oxide nanoparticles. Visualization of the introduced cell structures was performed using magnetic resonance imaging and ultrasound examination of the liver. 6 months after the cell therapy, clinical and biochemical blood tests, lidocaine test results, elastography indicators were evaluated, and the dynamics of hepatic encephalopathy was evaluated. The effectiveness of using autologous mesenchymal stem cells was evidenced by the improvement of the above indicators. Also, 6 months after treatment, a liver tissue biopsy was performed from the sites of fixation of previously introduced cell structures. Histological examination revealed the largest number of labeled cells in the liver micro-nodes, as well as at the borders of micro-nodes and fibrous septa. The use of cell therapy in a patient suffering from liver cirrhosis of alimentary etiology helped to improve the indicators of laboratory and instrumental research methods. No complications were detected during the procedure.

The purpose of the study is to evaluate the morphological and functional features of the tumor microenvironment and the cultural characteristics of mesenchymal stromal cells (MSC) of patients with multiple myeloma (MM) before and after treatment. Material and methods. Parenchymal and stromal bone marrow tissue in iliac bone trepanobiopsy of 32 patients with MM aged from 48 to 78 years (median age 56 years) before and after treatment was investigated. The study applied histological, immunohistochemical and cultural methods. Results and discussion. Сhanges of stromal microenvironment of bone marrow in all patients with MM have been revealed: increased microvascular density and the number of endosteal stromal cells, strengthening of reticulin fiber in subendosteal and perivascular spaces. Increased angiogenesis correlates with the number of plasma cells in the myelogram (r = 0.58; p < 0.05) and with the type of bone marrow infiltration (r = 0.85; р < 0.05), as well as with osteodestructive changes in the patient’s history (r = 0.65; p < 0.05). Significant changes in MSC cultures, both before and after therapy, including autologous hematopoietic stem-cell transplantation, were shown. Bone marrow MSC of patients with MM had decreased speed of proliferation (2.4 times on average), total time of passaging from 7 to 5 passages, increased expression of markers associated with myofibroblastic phenotype and aging (smooth muscle actin, β-galactosidase) compared to healthy donors. Conclusion. MSC cultures and BM niche forming elements of patients with MM have signs of tumor-associated microenvironment, despite the treatment of the underlying disease.

Objective: to visualize the cellular structures entered into the arterial course of a liver, for the purpose of assessment of a possibility of their fixing in liver tissue. Materials and methods . Introduction of mesenchymal stem autologichny cells, marked by iron oxide nanoparticles was carried out to 10 patients with the diagnosis cirrhosis. Introduction was carried out to the arterial course of a liver. Results. The mesenchymal autologichny cages entered into the arterial course of a liver, marked iron oxide nanoparticles, are visualized in tissue of a liver of patients by means of a magnetic and resonant tomography. Conclusion. Mesenchymal autologichny cages, marked iron oxide nanoparticles, it is possible to visualize in a human body by means of a magnetic and resonant tomography. The mesenchymal stem autologichny cells entered into the arterial course of a liver are fixed in liver tissue.