Влияние мультипотентных стромальных клеток, адсорбированных на полимере молочной кислоты, на воспалительную реакцию после его экспериментальной имплантации

Цель. Изучить влияние аутологичных мезенхимальных мультипотентных стромальных клеток (ММСК) костномозгового происхождения, адсорбированных на полимере молочной кислоты - полилактиде (ПЛ), на воспалительный процесс после экспериментальной имплантации этого полимера. Материалы и методы. С помощью световой микроскопии с применением люминесценции изучали изменения подкожно-жировой клетчатки крыс после имплантации ПЛ с пассивно адсорбированными на поверхности ММСК с трансфицированным геном GFP и дополнительно окрашенными Vybrant® CM-Dil клеточными мембранами. Результаты. Спустя 1 нед рядом с имплантированным ПЛ с адсорбированными ММСК методами флуоресцентной микроскопии были найдены фибробластоподобные клетки с ярким и равномерным свечением цитоплазмы при использовании родаминового фильтра. На 2-й неделе возле ПЛ присутствовали макрофаги разных размеров и форм, с интенсивной флуоресценцией множества включений в условиях применения родаминового фильтра. Далее яркость люминесценции и количество светящихся объектов прогрессивно снижались вплоть до почти полного исчезновения к 4-й неделе. В результате адсорбции на ПЛ ММСК уменьшается объем склерозированной клетчатки с увеличением в ней числа сосудов на 1 - 2-й неделях после имплантации. В этой клетчатке в течение 1-й недели численная плотность всех лейкоцитов и лимфоцитов ниже. Типичная капсула вокруг имплантированного ПЛ у крыс формируется только к 3-й неделе. Объем капсулы, ее васкуляризация и цитограмма лейкоцитов не зависели от адсорбции ММСК. Выводы. При имплантации ПЛ с адсорбированными ММСК уже к 2-й неделе эти клеточные элементы из тканей фагоцитируются макрофагами, ММСК и их детрит полностью элиминируются из места введения к 4-й неделе. Адсорбция ММСК на ПЛ способствует уменьшению выраженности склероза и воспалительных изменений подкожно-жировой клетчатки при увеличении ее васкуляризации на 1 - 2-й неделях после имплантации. Сроки формирования капсулы вокруг ПЛ, а также ее структура не связаны с использованием клеточных технологий

EVALUATION OF THE EFFICIENCY OF COMBINATION THERAPY WITH HYALURONATES IN PATIENTS WITH HIP OSTEOARTHROSIS

Objective: to make the results of treatment for coxarthrosis better via prosthetic synovial fluid replacement, by improving drug delivery into the joint cavity. Subjects and methods. The clinical trial enrolled 359 outpatients treated for coxarthrosis. A control group comprised 50 patients receiving the intraarticular combination drug Alflutop under ultrasound (US) guidance. A study group included 309 patients treated using a new three-stage procedure. At stage 1, the patients were given periarticular injections of the current enzyme preparation longidase with a hyaluronidase activity of 3000 IU. At stage 2, postisometric relaxation sessions were performed. At stage 3, after defining the optimal access, synovial fluid prosthetic devices were inserted into the joint cavity under US guidance (a total of 2-3 injections) once weekly. To evaluate the efficiency of the developed treatment option, the authors used a number of standard tests, WOMAC index, Leken's index, integral index of lower limb dysfunction, needs for nonsteroidal anti-inflammatory drugs; total treatment scores given by a physician and a patient; quality of life estimation using the SF-36 questionnaire. Results. The developed treatment option could significantly reduce the clinical and functional WOMAC index and Leken's index, and inadequate joint function in the immediate period and a year after the treatment performed. It also considerably improved quality of life in patients with hip osteoarthrosis, which appeared as a significant increase of its rating according to eight SF-36 scales during more than a year

Effect of soluble factors of macrophages polarized by efferocytosis on neuronal density in the frontal cortex and hippocampus of mice in a model of stress-induced depression

Recently, there has been a steady increase in depressive disorders, which occupy an important place in the structure of the causes of disability. In the pathogenesis of depression, an important role is played by neuroinflammation, which is associated with impaired adult neurogenesis. Notably, neuroinflammation is partially reversible, and the leading role in the initiation and regulation of neuroregeneration is given to macrophages. Opposite states of macrophage activation are classically activated M1 and alternatively activated M2 macrophages, characterized, respectively, by pro- and anti-inflammatory activity. A balance shift towards M2 macrophages has been considered as a new therapeutic strategy of psycho-neurological disorders. One of the inducers of the M2 phenotype is the efferocytosis. We have previously developed an original protocol for the generation of human macrophages under conditions of deficiency of growth / serum factors, in which M2 phenotype is formed through efferocytosis. Macrophages (M2(LS), LS – Low Serum) obtained according to this protocol express M2-associated markers, and are characterized by high production of growth and pro- angiogenic factors (IGF-1, VEGF, BDNF, EGF, FGF-basic, etc.), which can suppress inflammation and stimulate neuroregeneration / neuroplasticity. In the model of stress-induced depression, the antidepressant effect of soluble factors of M2(LS) macrophages was shown, accompanied by a decrease in the level of pro- inflammatory cytokines in certain brain structures. However, the effect of M2(LS) factors on neurogenesis remained unexplored. In the present work, which is a continuation of the aforementioned study, we analyzed the effect of intranasal administration of M2(LS) soluble factors on neuronal density in different brain areas – the frontal cortex and hippocampus – of depression-like mice. The results obtained showed that neuronal density in the frontal cortex, CA1 and CA3 zones of the hippocampus, was significantly higher in mice with intranasal administration of M2(LS) conditioned medium than in depression-like mice, and reached the level of neuronal density in intact animals. These results may indicate the neuroregenerative activity of M2(LS) macrophages in the model of stress-induced depression, which is mediated through soluble factors and manifests itself in an increase in the density of neurons in the brain

Neuromodulation possibilities in neurogenic lower urinary tract dysfunction

Introduction. Neuromodulation has proven itself in the treatment of patients suffering from idiopathic overactive bladder and non-obstructive urinary retention, who are resistant to conservative therapy. The possible use of the method in the population of patients with neurogenic lower urinary tract dysfunction (NLUTD) is of undoubted clinical interest.Objective. To analyze the current possibilities and features of neuromodulation in a cohort of patients with NLUTD.Materials and methods. Original research materials published in the PubMed, eLibrary, SciVerse (ScienceDirect), Scopus, Medline, EMBASE databases, websites of professional associations without restrictions on the date of publication were used. Sixty sources were selected for citation, with preference given to systematic reviews, meta-analyses and RCTs .Results. In relation to NLUTD, transcranial and peripheral magnetic stimulation, intravesical electrical stimulation, tibial, pudendal electrical stimulation, and stimulation of the dorsal pudendal nerve, as well as sacral and epidural methods of neurostimulation are considered.Conclusion. The current literature optimistically presents the experience of using neuromodulation in the NLUTD patient population with the largest evidence base for invasive sacral and tibial stimulation. The studies are based on heterogeneous populations, limited by small sample sizes with insufficient descriptive part of the degree and severity of neurological diseases, and it should be considered when forming guidelines. However, the lack of other suitable therapies and promising initial results indicate the importance of further efforts to improve the applied methods of neuromodulation. Further studies are needed with larger sample sizes, better classification of diseases, and controlled study desig

Некоторые особенности имплантации в условиях применения клеточных технологий

Некоторые особенности имплантации в условиях применения клеточных технологи

HUMAN BONE MARROW AND ADIPOSE TISSUE DERIVED MESENCHYMAL STROMAL CELL INFLUENCE ON NEUROLOGICAL DEFICIT RECOVERY IN A MODEL OF SEVERE TRAUMATIC BRAIN INJURY IN RATS

In the present study we characterized the effect of transplantation of bone marrow and adipose tissue MSCs in the model of brain injury in rats. This study was performed on Wistar rats (males and females in equal proportions with body weight of 250—270 g; n = 50). Traumatic brain injury was produced, by original spring-loaded, mechanism to precise dosing of impact force. Neurological disorders were assessed by Chen scale, a vertical grid test and a Morris water maze. MSC injections on day 1 were accompanied by a significant reduction in neurological deficit in compare with the control group. Adipose tissue derived MSCs were found to have a more pronounced effectiveness than MSCs obtained from bone marrow

Polymorphisms in the MTHFR and MTR genes and the risk of varicose veins in ethnical Russians

© 2016 Informa UK Limited, trading as Taylor & Francis Group.Objectives: The objective of this study was to study the association of polymorphisms MTHFR C677T (rs1801133) and MTR A2756G (rs1805087) with the risk of varicose veins in ethnical Russians. Methods: We genotyped 475 patients with varicose veins, 168 individual without chronic venous disease, and the population-based group of 896 subjects. Association was studied using logistic regression analysis adopting co-dominant, additive, recessive, and dominant models of inheritance. Results: None of the polymorphisms showed a statistically significant association with the risk of varicose veins. Conclusions: Our results provide evidence that the studied polymorphisms do not contribute to genetic susceptibility to varicose veins in ethnical Russians

MESENCHYMAL STROMAL CELLS IN CORRECTION OF NEUROLOGICAL DEFICIT AND MORPHOLOGICAL CHANGES IN THE RAT MODEL OF FOCAL CEREBRAL LESIONS INDUCED BY VENOUS BLOOD FLOW DISORDER

The paper presents the results of assessing the effect of bone marrow-derived mesenchymal stromal cells (MSCs) on the severity of neurological deficit and the brain morphology and function in a model of focal lesions induced by a venous blood flow disorder. Cerebral lesions were induced in Wistar rats by coagulation of the superior sagittal sinus followed by coagulation of cortical veins in the left parietotemporal region. MSCs were injected intravenously on days 1 and 7, and the dynamics of neurological disorders and morphological changes were assessed at 7, 14 and 21 days in comparison to controls. MSCs infusion was accompanied by a significant reduction. in neurological deficit, the most pronounced with MSCs injections on day 1. Morphological investigation of the damaged region have shown that the administration of MSCs led to a decrease in the area of necrosis, stimulation of angiogenesis, and improvement of structural and cellular parameters as compared to the control group. As a result, by day 21 the area of necrosis in animals with MSC transplantation was replaced by glial-mesodermal scar, whereas the scar formation in control animals was less pronounced, and in some cases was accompanied by cystic transformation. Transplantation of MSCs has a positive effect on neurological and morphofunctional recovery of the brain in experimental animal models of cerebral venous circulation disorders

Therapeutic effect of soluble factors of M2 phenotype macrophages in children with language impairments

The aim of the present study was to assess safety and clinical efficacy of inhalation immunotherapy based on intranasal administration of bioactive factors produced by the M2 phenotype macrophages in children with language impairments, as well as to study the effect of inhalation immunotherapy on the cytokine profile in the patients' blood serum. The study was carried out according to the NCT04689282 protocol (www.ClinicalTrials.gov) and included 14 children (9 boys / 5 girls), aged 3 to 8 years, with language impairments associated with perinatal or postnatal CNS lesions of various origin. The children recruited into the study were assessed by a neurologist and speech therapist before the therapy, at the end of the course (1 month), and 6 months later. Serum samples for cytokine analysis were obtained before and 1 month after therapy. The course of intranasal inhalations by the conditioned M2 media (2 ml one time per day for 28-30 days) was safe and well tolerated. None of the 14 treated children had significant adverse reactions and severe undesirable events. Intranasal immunotherapy led to a decrease in the severity of language problems, which manifested by improved speech understanding by 45%; the sensorimotor level of speech, by 51%; word formation skills, by 72%, as well as a twofold increase in general and fine motor skills. In children with signs of autism spectrum disorders, along with a language improvement, a decrease in the severity of autistic symptoms was registered, as evidenced by statistically significant decrease in the CARS score from 42.5 to 38.5 after 1 month, and to 33 points after 6 months (p < 0.05). The clinical effect was revealed rather soon, i.e., within a month after the first procedure, being maintained or intensified during a follow-up for 6 months. At the same time, two-thirds of the children showed a clear clinical improvement, with insignificant effect in the rest of patients. Comparative analysis of the serum cytokine levels in these subgroups showed that children with a pronounced positive response to inhaled immunotherapy differed in the following parameters: (1) initially higher level of VEGF and IGF-1, and (2) decrease the level of TNFα in response to intranasal immunotherapy. In summary, we first tested a fundamentally new approach based on the use of soluble factors from M2-type macrophages and intranasal route of their administration in order to treat the children with severe language impairments, demonstrating safety and obtained preliminary data on effectiveness of such approach

SPONTANEOUS AND LPS-INDUCED PRODUCTION OF 26 CYTOKINES SECRETED BY BLOOD CELLS OF PATIENTS WITH LIVER CIRRHOSIS DURING OF CELL THERAPY

The objective of the present study was to assess the level of 26 cytokines secreted by peripheral blood cells of the patients with liver cirrhosis (LC; n = 20) during the cell therapy (CT). All the patients were administered with intravenously injected autologous bone marrow-derived mononuclear cells (MNCs) in a dose of 1.3±0.3 × 109 (Ме 1.0 × 109 ) followed by 14 days later intravenous injection of ex vivo generated mesenchymal stromal cells (MSCs) in a dose of 22.3±5.0 × 106 (Ме 16.0 × 106 ). The patients were examined before the CT, 2-3 days after the administration of MNCs and, then, 2-3 days after the introduction of MSCs. Cytokine-secretory function of peripheral blood cells was evaluated in a 24-hour whole blood cultures both in the absence of any stimulation and in response to lipopolysaccharide (LPS). The control group consisted of 10 healthy donors. The administration of patients’ bone marrow cells (both MNCs and MSC) was safe and well tolerated and caused no any adverse (toxic or allergic) events. Compared with donors, LC patients (especially, with class B+C by Child-Pugh) differed by an initially increased production of several cytokines and chemokines. Actually, there was a statistically significant increase of the spontaneous production of IL-9, MIP-1β, and IP-10, as well as a distinct trend to an increase in TNFα, IL-1ra, IL-4, IL-5, IL-6, IL-13, of MCP-1, MIP-1α, RANTES and Eotaxin. Moreover, the blood cells of LC patients were susceptible to the stimulatory effect of LPS, and the LPS-induced production of 11 out of 26 cytokines (IL-1ra, IL-6, IL-9, IL-15, IL-17, IL-7, IL-8, IP-10, MIP-1α, MIP-1β, Eotaxin) significantly exceeded the normative values. Transplantation of bone marrow MNCs had minimal impact on cytokine production. Meanwhile, the MSCs introduction resulted in a significant decrease in spontaneous and LPS-induced production of, respectively, 20 and 18 analytes including pro-/anti-inflammatory and immunoregulatory cytokines, chemokines and growth factors. The normalization of cytokine-secretory function following transplantation of MSCs revealed in the patients with LC indicates the weakening of an inflammatory activity of circulating blood cells and the decrease in their reactivity to endotoxin. MSC suppressive effect on cytokine production was dose-dependent, and most pronounced in patients with decompensated LC (class B+C by Child-Pugh) of viral etiology