Pathogenetic rationale for the use of cell therapy in lung injury associated with SARS-CoV-2

Acute respiratory disease COVID-19 caused by the SARS-CoV-2 coronavirus demonstrate weak clinical manifestation in most patients. However, pneumonia and acute respiratory distress syndrome in some cases may cause serious problems due to the lack of effective etiotropic and pathogenetic therapy. Presumably, SARS-CoV-2 leads to the delayed type I interferon activation and loss of control over virus replication in the early stages of infection, which is why the adaptive CD8+T-cell response must be controlled to avoid the development of pulmonary pathology. These data should be taken into account when developing strategies for COVID-19 therapy. Mesenchymal stem cells therapy serves as possible treatment opportunity for severe forms of the disease due to their homing, pronounced anti-inflammatory and antifibrotic properties. It was found that in viral infections, including COVID-19, mesenchymal stem cells can synthesize antiviral defense mediators under the influence of interferon causing resistance to viruses. Thus, mesenchymal stem cells are able to provide comprehensive anti-inflammatory protection, which leads to clinical improvement in patients with COVID-19

Diagnostic power of laboratory methods for assessing ulcerative colitis severity: A prospective comparative study

Background. The epidemiology of ulcerative colitis in the Russian Federation is typified by late diagnosis and the predominance of severe complications entailing high mortality.Objectives. A comparison of the diagnostic value of laboratory methods for assessing ulcerative colitis severity.Methods. A total of 178 ulcerative colitis patients were divided into 4 cohorts by the presence and severity of attack; a control cohort included 40 healthy volunteers. Besides standard tests, a cytokine profile was determined in all patients: IL-1 IL-2, IL-4, IL-6, IL-10, TNF-α, IL-17; faecal inflammation marker concentrations — lactoferrin (LF), calprotectin (CalP), neopterin (NP); optical anisotropy (OA) of neutrophilic granulocyte nuclei; clinical activity index (CAI); Mayo scores. A single-factor analysis of variance was performed to compare the diagnostic value of laboratory tests, with a Mayo score taken as the factor reflecting ulcerative colitis attack severity. Differences were assumed statistically significant at p < 0.05. Results. The patients suffering from ulcerative colitis exhibited statistically significant Mayo score correlations with IL-6 (r = 0.598, p = 0.001), IL-17 (r = 0.587, p = 0.005), TNF-α (r = 0.701, p = 0.001), CalP (r = 0.881, p = 0.001), LF (r = 0.799, p = 0.001), NP (r = 0.791, p = 0.001) and OA (r = –0.877, p = 0.001). Faecal inflammatory biomarkers varied in the range from 73.4 (NP) to 95.3% (CalP) of total variance. Serum markers varied from 75.2 (IL-6) to 88.1% (IL-17) of total variance. As of all markers, the highest diagnostic value was observed for CalP (95.3% of total variance), while the lowest — for NP (73.4% of total variance). In the analysis of variance, the cut-off values for serum markers in predicting endoscopically active disease (>1 Mayo score) comprised: IL-6 = 10.3 pg/mL; IL-17 = 18.5 pg/mL; TNF-α = 10.9 pg/mL. The analogous values for faecal markers were: CalP = 112.0 μg/g; LF = 80.9 μg/g; NP = 92.8 μg/g. Neutrophilic granulocytes optical anisotropy comprised 94.5% of total variance, which compares with CalP by diagnostic power.Conclusion. A high diagnostic power has been demonstrated for faecal inflammatory markers (calprotectin, neopterin, lactoferrin), cytokines (IL-6, IL-17, TNF-α) and neutrophilic granulocytes optical anisotropy in detecting the attack relapse and severity

Surgical aspects of full-thickness skin autograft engraftment on a granulating wound

Background. The paper presents the clinical results of treating patients using full-thickness skin autografts for granulating wounds.Objective. To study the surgical aspects of full-thickness skin autograft engraftment on a granulating wound.Material and Methods. In traumatology and burn research, to achieve the best cosmetic results, transplantation methods of free full-thickness skin autografts are used. In the Regional Clinical Hospital no. 1, a method of skin grafting with a full-thickness autograft was developed, which allows to close the defect in the conditions of a purulent wound: granulations are excised before skin transplantation, and a vacuum is applied after grafting.Results. Annually, the specialists of the Burn Center of the Scientific Research Institute – Regional Clinical Hospital no. 1 assist more than 1300 patients. Of these, from 20 to 25 cases are face burns. 132 patients with burns of the face have been admitted to the hospital over the past 10 years. 37 patients underwent plastic surgery with a full-thickness skin autograft. The authors presented the technology for the surgical treatment of deep burns on the face. After cleansing the wound from areas of necrosis and granulations, usually 20 days after the injury, the upper layers of granulation tissue are removed with a dermatome parallel to the skin surface, to a depth of 1–2 mm. Then, plastic surgery with the free full-thickness skin autograft is carried out on the skin of the face with the formation of cuts for the nose, mouth, eyes. The wound is tightly bandaged with 5–7 rounds of a medical bandage or a vacuum-assisted closure is applied.Conclusion. In case of traumatic skin detachment, plastic surgery according to Krasovitov should be performed in the first hours from the moment of injury. Our study allows transplantation of a full-thickness skin graft to granulating wounds as well. After 5 days, the condition of skin autografts is assessed at the first dressing. Their engraftment is observed on the 7th day. In the postoperative period, scar tissue does not form

Application of cells of cord blood and umbilical cord: achievements, challenges and perspectives

This review focuses on the biological aspects of the use of cord blood as a valuable source of cells. The distinctive features of hematopoietic stem cells, cells of the immune system, mesenchymal stem cells are described. The analysis of the results of clinical research and development of therapeutic approaches using cord blood cells and umbilical cord for the treatment of various diseases has been carried out.Currently, the target area of cord blood research is hematopoietic stem cell transplantation, as well as cellular immunotherapy of tumor diseases, treatment of neurological diseases and regenerative medicine

Characterization of Stem-Like Cells in Mucoepidermoid Tracheal Paediatric Tumor

Stem cells contribute to regeneration of tissues and organs. Cells with stem cell-like properties have been identified in tumors from a variety of origins, but to our knowledge there are yet no reports on tumor-related stem cells in the human upper respiratory tract. In the present study, we show that a tracheal mucoepidermoid tumor biopsy obtained from a 6 year-old patient contained a subpopulation of cells with morphology, clonogenicity and surface markers that overlapped with bone marrow mesenchymal stromal cells (BM-MSCs). These cells, designated as MEi (mesenchymal stem cell-like mucoepidermoid tumor) cells, could be differentiated towards mesenchymal lineages both with and without induction, and formed spheroids in vitro. The MEi cells shared several multipotent characteristics with BM-MSCs. However, they displayed differences to BM-MSCs in growth kinectics and gene expression profiles relating to cancer pathways and tube development. Despite this, the MEi cells did not possess in vivo tumor-initiating capacity, as proven by the absence of growth in situ after localized injection in immunocompromised mice. Our results provide an initial characterization of benign tracheal cancer-derived niche cells. We believe that this report could be of importance to further understand tracheal cancer initiation and progression as well as therapeutic development

CLINICAL EFFECTS OF MESENCHYMAL STROMAL CELLS IN LYMPHOMA PATIENTS WITH AUTOLOGOUS HEMATOPOIETIC STEM CELL TRANSPLANTATION

The clinical and laboratory effects of bone-marrow derived mesenchymal stromal cells (MSC) in patients with malignant lymphomas following autologous hematopoietic stem cell transplantation (auto-HSCT) have been investigated. Co-transplantation of MSC in average dose of 0,178 106/kg was conducted in 74 patients with auto-HSCT. The control group included 83 patients eligible for standard HSCT. We revealed the decreasing of the period of neutropenia and thrombocytopenia when hematopoietic stem cells were co-transplanted with low doses ex vivo expanded autologous MSC. Patients with MSC co-transplantation were differed by more effective early lymphocyte recovery. At the same time MSC co-transplantation did not increase the incidence of infectious complications and cases of renal and. hepatic toxicity. Patients with MSC co-transplantation did not differ from opposite group by 5-year overall survival, but were characterized by significantly better progression-free survival

THE COMPARATIVE ANALYSIS OF GRAFT CELL SUBTYPES AND ITS CYTOKINE PRODUCTION OF LYMPHOMA AND LEUCOSIS PATIENTS

Cell subtypes and cytokine profile of apheresis products collected from lymphoma and acute leucosis patients were analyzed. It was shown, that acute leucosis patients' grafts contained higher relative numbers of naïve T-cells, CD4+CD25high T-cells, T-lymphocytes (non-significant trend) and lower counts of granulocytes. Significant increase of relative numbers of dividing CD34+ cells (in S, G2/M phases of the cell cycle) was demonstrated, in acute leucosis patients' grafts. In lymphoma grafts the levels of CD34+ cells in G0/G1 phases were found, to be increased. Cells isolated from grafts of acute leucosis patients characterized by higher levels of proinflammatory cytokines production, such as IL-1, IL-6, MIP-1β, TNF-α, IL-8, IFN-γ (the last two ones— non-significant trend) and cytokines, essential for humoral immune response (IL-4 and — in trend — IL-10). The existing differences didn't influence on effectiveness of early lymphocyte recovery

ОПЫТ ПЕРФУЗИОННОЙ РЕЦЕЛЛЮЛЯРИЗАЦИИ БИОЛОГИЧЕСКОГО КАРКАСА ЛЕГКИХ КРЫСЫ

Aim. The main aim of our research is to evaluate the process of rat lung decellularization and recellularization as the initial step of tissue-engineered organs creation.Materials and methods. Rat lung decellularization was performed by perfusion with detergents and enzymes with concomitant atmospheric air ventilation through the trachea. The quality of decellularization was analyzed with routine histological and immunohistochemical staining techniques, DNA content was determined quantitatively by spectrophotometer. For static and whole organ reseeding as a model of cells’ behavior mesenchymal multipotent stromal cells were used. Recellularization was followed by assessment of the cellular metabolic activity by colorimetric method; cell viability was analyzed by calcein and ethidium homodimer staining. Matrix qualitative evaluation after recellularization was performed using immunohistochemical staining methods.Results. 92% of allogeneic DNA was eliminated after decellularization. Histological staining revealed no residual cells and cell nuclei; preservation of the fibers of extracellular matrix was confirmed by immunohistochemical staining for laminin, elastin, fibronectin, collagen types I and IV before and after decellularization. The scaffold does not exhibit toxic properties after reseeding; cell viability and metabolic activity were proved after cultivation.Conclusion. The experience of rat lung decellularization and recellularization can be the prospective basis for protocols of organ recellularization and tissue engineered lungs creation.Цель исследования – оценить перспективы использования процессов децеллюляризации и рецеллюляризации легких крысы как начального этапа создания тканеинженерных конструкций.Материалы и методы. Децеллюляризацию легких крысы выполняли перфузионным детергент-энзиматическим методом при сопутствующей вентиляции трахеи атмосферным воздухом. Качество проведенной децеллюляризации оценивали с использованием рутинных гистологических и иммуногистохимических методов исследования, количественное содержание ДНК определяли спектрофотометрически. Для статической рецеллюляризации и перфузионной рецеллюляризации целого органа в качестве модели для изучения поведения клеток на каркасе использовали мезенхимальные мультипотентные стромальные клетки с последующей оценкой метаболической активности клеток колориметрическим методом и их жизнеспособности – путем окрашивания кальцеином и гомодимером этидия. Для качественной оценки матрикса легких после рецеллюляризации использовали иммуногистохимический анализ.Результаты. 92% аллогенной ДНК удалено при проведении децеллюляризации. Гистологическое окрашивание не выявило остаточных клеток и клеточных ядер при сохранности волокон внеклеточного матрикса, что подтверждалось иммуногистохимической реакцией матрикса с антителами к ламинину, эластину, фибронектину, коллагенам I и IV типов до и после проведения децеллюляризации. Каркас при заселении клетками не проявляет токсических свойств, поддерживает жизнеспособность и метаболическую активность клеток.Заключение. Полученный опыт децеллюляризации и рецеллюляризации легких крысы является перспективной основой для разработки протоколов создания тканеинженерных легких

Разработка методики получения дермального внеклеточного матрикса

Despite advancements in modern surgery in the treatment of cutaneous injuries, the search for new methods of ensuring faster and more effective wound healing appears especially urgent today. Tissue engineering is undoubtedly of interest when it comes to developing such technologies. Objective: to determine the optimal protocol for obtaining a decellularized dermal matrix scaffold for subsequent development of tissue-engineered skin. Materials and methods. One Landrace piglet was used as the experimental animal. After preliminary skin treatment with dermatome, 0.3 cm thick samples were taken. Two decellularization protocols were considered: protocol No. 1 was based on the use of Triton X-100 and deoxycholate, protocol No. 2 was only based on deoxycholate. There were 5 processing cycles in total for the 2 protocols. After treatment, acellular matrix scaffolds were examined through histological analysis and quantitative determination of DNA concentration. Next, static recellarization of the matrix scaffolds was carried out with porcine dermal fibroblasts. After that, the matrix scaffolds were tested for cytotoxicity using XTT test and differential staining test to differentiate between live and dead cells. Results. Comparative analysis of the two protocols for porcine dermis decellularization showed that both protocols effectively remove cells and nuclear material, while maintaining the architectonics of the intercellular substance intact, since fibrous structures are not destroyed. But when assessing the biocompatibility of matrix scaffolds based on analysis of cell viability according to data obtained from XTT test and cell–matrix adhesion, the matrix scaffold processed under protocol No. 1, shows advantages. Conclusion. In this study, a decellularization protocol based on Triton X-100 and deoxycholate was noted. The results obtained mark the first stage towards developing a tissue-engineered skin.Несмотря на достижения современной хирургии в лечении повреждений кожных покровов, актуальным остается поиск новых методов для более быстрого и эффективного заживления ран. Тканевая инженерия, несомненно, представляет интерес для разработки таких технологий. Цель данной работы состояла в определении оптимального протокола получения децеллюляризированного дермального матрикса для последующей разработки тканеинженерной кожи. Материалы и методы. Экспериментальным животным был 1 поросенок породы Ландрас. После предварительной обработки кожи дерматомом забирали образцы толщиной 0,3 см. В работе рассматривалось 2 протокола децеллюляризации: протокол № 1 на основе применения тритон Х100 и дезоксихолата, протокол № 2 только на основе дезоксихолата. Всего циклов обработки по 2 протоколам было 5. Ацеллюлярные матриксы после обработки были исследованы следующим образом: гистологический анализ, количественное определение содержания ДНК. Далее была проведена статическая рецеллюляризация матриксов фибробластами дермы свиньи. После чего матриксы были исследованы на цитотоксичность с помощью ХТТ-теста и теста на дифференциальное окрашивание живых и погибших клеток. Результаты. Проведенный сравнительный анализ двух протоколов децеллюляризации дермы свиной кожи показал, что оба протокола эффективно удаляют клетки и ядерный материал, при этом сохраняется архитектоника межклеточного вещества неповрежденной, так как не происходит разрушения волокнистых структур. Но при оценке биосовместимости матриксов на основе анализа жизнеспособности клеток по данным ХТТ-теста и адгезии клеток к матриксу преимущества демонстрирует матрикс, обработанный по протоколу № 1. Заключение. В настоящем исследовании был отмечен протокол децеллюляризации на основе тритон Х100 и дезоксихолата. Полученные результаты являются первым этапом для дальнейшей разработки тканеинженерной кожи