Objective: The dualistic role of IL-6 in diseases associated with metabolic disorders (including non-alcoholic fatty liver disease, NAFLD) under oxidative stress has been repeatedly demonstrated in various studies. The aim of this work was to investigate the combined effect of IL-6 and glycoprotein 130 (gp130) on cell death pathways and mitochondrial dynamics in the HepG2 cell line in the oxidative stress model. Methods: The HepG2 cell line was used to create a model of oxidative stress in hepatocytes (hepatocytes without any additives are hereafter referred to as control) by adding tert-butyl hydroperoxide (tBHP). IL-6 (variant 1) or IL-6 + gp130 (variant 2) were added to the experimental model of oxidative stress. Cell viability was determined by flow cytometry and the expression and production of key markers of mitochondrial biogenesis, apoptosis, mitophagy and antioxidant defense were analyzed by RT-qPCR and Western blotting, respectively. Results and Discussion: The addition of IL-6 alone (variant 1) increased the relative percentage (

Introduction. Artificial materials used in regenerative medicine induce a balanced inflammatory response after implantation, which is an important step for effective regeneration of damaged bone tissue. The contact of the implant with tissues and biological fluids is accompanied by the deposition of blood proteins on its surface, which contributes to the activation of the complement system and initiates blood clotting, leading to the formation of a fibrin clot. On the surface of the implant, fibrin ensures the adhesion of stem cells and their maturation into fibroblasts that produce collagen and its derivatives. The formed extracellular matrix is the basis for the formation of a tissue structure (callus). To prevent the development of postoperative pathological conditions caused by hypercoagulatory syndrome, therapeutic strategies with anticoagulants such as heparin are used. However, their use limits the formation of a fibrin clot in vivo, which may slow down the migration of mesenchymal stromal cells (MSCs) and the subsequent formation of callus. Aim. To investigate of the effect of heparin at pharmacological concentrations on stemness and the ability of MSCs from human adipose tissue to undergo osteogenic differentiation under conditions of in vitro cultivation. Materials and methods. To assess the morphofunctional state of cells cultured in the presence of heparin, 2 experimental groups were formed: 1) MSCs in the presence of heparin at a therapeutic concentration (1.3 IU/ml); 2) MSCs in the presence of heparin at a toxic concentration (13 IU/ml). Results and discussion. Flow cytometry results showed that the addition of heparin at both concentrations used in the study to MSC culture leads to an increase in the number of cells expressing the surface markers CD73 and CD90, indicating the maintenance of their stem state. On the other hand, a stimulatory effect of heparin at both concentrations used on the transcription of mRNA of osteogenic genes (BMP2, BMP6, ALPL, RUNX2, BGLAP and SMURF1) in MSCs was also observed, which may indicate the osteogenic potential of heparin for the cell culture studied. Conclusion. The results of the study are useful for regenerative medicine related to the use of MSCs in clinical practice; they may serve as a prerequisite for the development of new therapeutic strategies for orthopedic and traumatologic patients at high risk of postoperative thrombosis after endoprosthetics surgery and osteosynthesis.

Autophagy is required to maintain cellular homeostasis and organ function by selectively ridding cells of potentially toxic proteins, lipids, and organelles. Impaired homeostasis of autophagic processes is associated with metabolic disorders such as obesity and type 2 diabetes mellitus. In obesity, a violation of autophagy in adipose tissue and its inflammation contributes to the formation of type 2 diabetes mellitus. The aim of the study was to analyze the expression of autophagy genes in the adipose tissue of the greater omentum and to search for their relationship with the levels of cytokines of the IL-10 family in blood plasma in obese patients, depending on the presence or absence of type 2 diabetes mellitus. Blood plasma and visceral adipose tissue samples were studied from 347 obese patients with and without type 2 diabetes. A biochemical analysis of the patients' blood was carried out. The level of cytokines was detected by flow fluorometry. Gene expression was determined by real-time PCR, and tissue-specific protein production was determined by immunoblotting. Statistical processing of the results was carried out using GraphPad Prism 9.0.0 software. Plasma levels of IL-10, IL-20, IL-22, IL-28A, and IL-29 are increased in obese patients without type 2 diabetes compared with patients with type 2 diabetes. In patients with type 2 diabetes mellitus, the expression of the SQSTM1_p62 and MAP1LC3B genes in the greater omentum increased compared to patients without it. High plasma levels of IL-22 and IL-26 are associated with the presence of type 2 diabetes mellitus. In patients without type 2 diabetes mellitus, an increase in the level of IL-28A in blood plasma is associated with a decrease in the expression of autophagy genes SQSTM1_p62 and MAP1LC3B in the adipose tissue of the greater omentum.

The article is devoted to studying the role of mesenchymal stromal cells in formation of microenvironment for hematopoietic stem cells under the conditions mimicking physiological bone remodeling in presence of artificial three-dimensional matrices (Ra = 2-3 μm). The study was carried out using experimental samples of artificial implants obtained in electrolyte from hydroxyapatite nanoparticles (HAP) produced at the Institute of Strength Physics and Materials Science (Siberian Branch of the Russian Academy of Sciences). The work included cultural and instrumental research techniques. Phenotypic profile of cells was assessed by flow cytometry. Determination of cytokine/chemokine levels from cell culture supernatants was assessed by flow fluorimetry. Detection of hematopoietic cells in the vision fields, as well as areas of extracellular matrix mineralization was carried out by means of cytomorphometry.It was revealed that the 3D matrices with a calcium phosphate coating initiate the in vitro formation of specific microenvironment of MSCs, resulting in the increased numbers of HSCs with the CD45+CD34+ phenotype (at 14 days), an increased number of cells with hematopoietic morphology and evolving foci of extracellular matrix mineralization of the (at 21 days). Changed numbers of hematopoietic cells per vision field occurred, mainly, due to indirect effect of hematopoietic factors (SCF and G-CSF), along with decrease of proapoptotic factor TRAIL. It was also found that MSCs reduce the level of proinflammatory cytokines (IFNγ, TNFα, IP-10, IL-2, IL-6) in culture medium in the presence of artificial 3D calcium-phosphate-coated matrices. The revealed features of MSC functioning under the conditions simulating physiological bone remodeling, upon co-cultures with three-dimensional matrices (Ra = 2-3 μm), have shown a significant effect of MSCs upon regulation of HSCs by local microenvironment, through distinct modulating effects of cytokines, chemokines, and growth factors that provide intercellular interactions. Development of extracellular matrix mineralization areas during MSC cultivation in the presence of 3D matrices imitating mineral substance of bone tissue also indicates the formation of osteoblastic niches under the in vitro cultivation conditions.The results obtained are important in order to assess functions of hematopoietic niches and the role of MSCs in their development and maintenance of the microenvironment.The results obtained may find practical application in development of new classes of medical devices able to provide effective osseointegration.

Correct choice of nutrient media for culturing different types of cells in various applications is one of the most important aspects of modern biotechnology, since chemical composition of the culture media largely contains the necessary metabolites to support certain cells’ growth lines outside the body. Jurkat line of human leukemic T-lymphoblast-like cells (hereinafter Jurkat T-cells) is actively used for in vitro modeling of intracellular signaling and activation of normal blood T-lymphocytes mediated by the T-cell receptor/CD3/ CD4 complex in toxicological studies of immune and secretory responses, to test medicinal substances and ions. Also, Jurkat T-cells are widely used for ex vivo testing in immunology, oncology, toxicology, orthopedics, and traumatology. The existing standards and numerous studies are mainly based on short-term in vitro cultivation of Jurkat T-cells in RPMI 1640 nutrient medium. Meanwhile, the issues of long-term maintenance of the growth of Jurkat T-cells culture are poorly presented in the research literature. This study aimed for studying the activity of Jurkat T-cells over 7 to 14 days of in vitro culture and comparing the relative value of RPMI 1640 and αMEM media for the behavior of immunocompetent tumor cells. Using flow cytometry, multiplex analysis, and phase contrast Cell-IQ microscopy, the proportions of living cells and those dying by apoptosis and necrosis, secretion of cytokines and chemokines, and the dynamics of cell biomass propagation were studied. It was found that the αMEM medium in the complete nutrient medium, as compared with RPMI 1640, is more appropriate to in vitro promotion of cell viability (increased proportion of viable cells by 13.5% at the day 14), their secretory ability for 23 из 27 tested biomolecules, shortened adaptation time (на 32%) in culture before growth initiation, 5-fold increase of the Jurkat Т-cell cellularity by the day 7. Potential significance of the chemical components of nutrient media and secreted biomolecules for these results is discussed. As based on the results obtained, we concluded on superior properties of αMEM medium for long-term in vitro cultures of Jurkat T-cells. Consequently, the in vitro testing of medical devices intended for long-term contact with the body, including those for cancer patients, using Jurkat T-cell leukemia line in RPMI 1640 medium, may lead to wrong predictions on their biocompatibility and potential antitumor activity.

In recent years, materials based on graphene oxide (GO) have been actively studied for their use in biomedicine. The aim of our study was to investigate the increase in cell mass and viability of Jurkat tumor line T cells during 24 h of contact with GO nanoparticles in the Cell-IQ system of intravital observation. We used nanoparticles of different sizes coated with linear or branched polyethylene glycol (PEG) at concentrations of 5 and 25 μg/mL. It was shown for the first time that direct contact with GO nanoparticles reduced the growth in cell mass at the visualization points by more than twofold, regardless of nanoparticle size and concentration. Moreover, the number of live cells in the culture decreased by 5–9% after 24 h of monitoring. Thus, PEG-coated GO nanoparticles were found to suppress the proliferation and viability of Jurkat cell line T lymphocytes.

Molecular genetic mechanisms, signaling pathways, cultural conditions, factors, and markers of osteogenic differentiation of mesenchymal stem cells (MSC) are actively studied despite numerous works in this area of cellular technologies. This is largely due to the accumulating contradictions in seemingly classical knowledge, as well as permanent updating of the results in the field. In this regard, we focused on the main classical concepts and some new factors and mechanisms that have a noticeable regulatory effect on the differentiation potential of postnatal MSCs. The present review considers the significance of MSC sources for their differentiation capacity, as well as the role of the cellular microenvironment. The issues of classification, terminology, and functional activity of MSCs from various sources are discussed. The paracrine potential of MSCs in tissue regeneration has been considered; sufficient importance of inflammation in osteogenesis is noted, in particular, the presence of inflammatory cytokines and chemokines in the lesion focus, produced not only by microenvironmental cells but also by blood cells, including mononuclear leukocytes, migrating to the affected site. An important role in this review is given to biomechanical signals and to influence of conformational changes in cell cytoskeleton (cell shape) upon MSC differentiation, since the morphological features of cells and the structure of cytoskeleton are modulated by interactions of the cell surface with environmental factors, including hydrostatic pressure, fluid flow, compression/stretching loads. The data are presented concerning elasticity of extracellular matrix being a determining factor of cell differentiation. We conclude that one should switch from point studies of individual gene effects to multiple measurements of the gene-regulatory profile and biomolecules responsible for multiple, still poorly studied osteogenic factors of endogenous and exogenous origin. Among cornerstones in future (epi)genetic studies will be to decide if osteomodulatory effects are realized through specific signaling pathways and/or via cross-signaling with known genes controlling osteogenic differentiation of MSCs.

T-lymphoblast-like human leukemia cells of the Jurkat line (Jurkat T cells) form polyploid forms with a high DNA content in suspension culture. Due to pronounced genetic instability, this contributes to further transformation and development of clonal diversity (polyclonal) of the cell line. Little information is available on the adherent subpopulation of Jurkat T cells. In this work, we analyzed the content of nucleic acids in suspension (DNA) and adhesive (DNA, RNA) subpopulations of Jurkat T cells using flow cytometry and propidium iodide dye, as well as confocal laser microscopy and acridine orange dye. The morphology and mobility of large (with a diameter of more than 15 μm) Jurkat T cells adhering to plastic were studied using Cell-IQ phase-contrast microscopy in real-time. According to the intensity of fluorescence in the conditionally green wavelength range (300–530 nm: from UV to green) and conditionally red (565–800 nm: from red to far-red), three subpopulations of adherent Jurkat T cells were identified: with high, medium, and low nucleic acid content. Thus, Jurkat-T cells adhering to the plastic surface of the plates retain a pronounced heterogeneity in the DNA content characteristic of the suspension fraction, which suggests a difference in the morphofunctional properties (polyclonicity) of this subpopulation of cell culture. With a sharp increase in the total cell mass, the proportion of large (giant, 15–50 μm or more) cells attached to the plastic remained constant for 21 days of cultivation and amounted to 1% of the adhesive fraction. It was found that large Jurkat T cells (with a median diameter of 31 µm) moved along the plastic at a linear (median) speed of 38 µm/h. The polynuclear Jurkat T cells on plastic are morphologically identified, with a linear increase being revealed in the mobility of adherent cells with an increase in their diameter (regression coefficient r = 0.33, p < 0.02, n = 52). Possible cellular and molecular mechanisms of an increased number of DNA copies in some adhering Jurkat T cells are discussed. It is assumed that the discovered new property (locomotor activity) can provide polyploid (multinucleated) adhering Jurkat T cells with a significant advantage—directed migration (chemotaxis) in a growing cell population under conditions of nutrient deficiency due to a change in the nutrient medium after 3–4 days of cultivation.

Synthetic materials used in regenerative medicine, upon implantation, induce the development of an inflammatory reaction necessary for the effective regeneration of damaged bone tissue. Implant contact with tissues is accompanied by the deposition of blood proteins and interstitial fluid on its surface, contributing to the activation of the complement system, components of innate immunity, initiating coagulation hemostasis, leading to the formation of a fibrin clot. An extracellular matrix based on fibrin, collagen and elastin forms on the implant’s surface, which provides the basis for the formation of tissue structure through the adhesion of stem cells to the forming bone callus before the formation of bone regenerate. To prevent the development of postoperative pathological conditions caused by hypercoagulable syndrome, therapeutic strategies are used to use anticoagulants (heparin, warfarin). However, their use limits the normal formation of a fibrin clot in vivo. This can slow down the migration of mesenchymal stem cells (MSC) and disrupt the formation of callus, inhibiting the processes of osseointegration of the implant and bone healing. The study’s goal was to study the effect of heparin in a gradient of low and high concentrations on the migration activity and stem capacity of human MSCs under in vitro cultivation conditions. According to the results of flow cytometry, it was revealed that high concentrations of heparin (130, 260 IU/ml) in a 2D cultivation model contribute to an increase in the number of cells expressing surface markers CD73 and CD90, which indicates that MSCs retain high clonogenic potential. A 3D model of in vitro cultivation with the addition of heparin and osteosubstituting implants bearing a CF coating with a roughness index of Ra = 2.6-4.9 μm contributed to preserving the “stemness” character of MSCs through the expression of surface markers CD73 and CD90. According to the results obtained using the xCELLigence system, heparin at a later time (from 20-40 hours) increases the invasion of MSCs through micropores that simulate the state of the blood vessel walls. However, in the presence of HAP nanoparticles that mimic the remodeling processes of the mineral bone matrix and/or resorption of bone cement, the effect of heparin was less pronounced. The results can be used in the field of regenerative medicine associated with the introduction of MSCs. The data can serve as a prerequisite for developing new therapeutic strategies for surgical patients with a high risk of postoperative thrombosis after osteosynthesis.

The review considers complex, controversial, and individual effects of heparin and its derivatives on the bone and circulatory systems in dependence of the dose, the state of the cells and tissues of the recipient. General data on the anticoagulant activity of heparin and its derivatives are presented; special attention is paid to the effect of heparin on mesenchymal cells and tissues and its role in angiogenesis. We also discuss the ability of heparin to bind osteogenic and angiogenic biomolecules in the context of the development of systems for their delivery and sustained controlled release and propose a schematic representation of the positive and side effects of heparin as a delivery system for biomolecules in tissue engineering.

Currently, there is an increasing demand for biocompatible materials that can be used for bone reconstruction. However, there is still no consensus regarding adequate bone replacement material. The materials traditionally used for reconstructive surgeries, and methods of making bone-replacing implants from them have various disadvantages. They do not fully satisfy the biological and biomechanical characteristics of living tissues. This leads to a clinical situation called "implant failure” and consists of a violation of its integrity, loosening, attachment of infectious agents, and inflammation development. There are severe socio-economic losses for the patient himself and the state. The problem of infectious complications after surgical operations with the use of bone replacement implants is quite acute. Periprosthetic infection is a modern professional challenge for surgeons and bioengineers. However, antibiotic therapy, which is the only treatment of choice for periprosthetic infection, is characterized by various side effects and becomes ineffective due to microbes' antibiotic resistance. In this regard, for the fight against periprosthetic infection, metal ions with antimicrobial potential (copper, zinc) are considered promising, which are not destroyed during sterilization of medical devices and have their own biological (regulatory) activity. The presented data indicate researchers' interest in studying the interaction of immunocompetent and mesenchymal stem cells with biomedical materials with antimicrobial potential.

Osteoimmunolgy describes the interactions between blood immune cells and human multipotent mesenchymal stromal cells (hMMSCs) as a basis of successful wound and fracture healing. The aim was to investigate in vitro interaction of adipose-derived hMMSCs and human blood mononuclear cells (hBMNCs) modulated by calcium phosphate (CaP) coating in 3D culture. Titanium plates (10 × 10 × 1 mm3) with micro-arc bilateral multilevel CaP coatings were used. Cell-IQ phase-contrast microscopy and the real-time cell analyzer (RTCA) showed the CaP coating and hBMNCs synergistic negative effects on hMMSC motility that could be conditioned by enhanced osteogenic differentiation of stromal cells. Indeed, a 10-fold increase in the bone mineralization around the CaP-coated samples was detected in the mixed (hBMNCs + hMMSCs) 3D culture. Thus, cellular and molecular crosstalk between hBMNCs and hMMSCs modulated by multilevel micro-arc CaP coating with bone-like topography is an effective 3D model to study in vitro the novel pathways of osteoimmunology.

Introduction: Currently, traumatology actively uses ceramic materials with calcium phosphate coating (CP). Such materials have a stimulating effect on multipotent mesenchymal stromal cells (MMSCs) and immunocompetent cells. So, their use involves an assessment of the impact of these materials on the development of tumor cells. Methodology: The Jurkat 5332 cell line and human adipose-derived MMSCs (AMMSCs) were examined. 3D culture was simulated by adding to the cell culture the substrates from commercially pure titanium with rough (Ra=2-5 µm) CP microarc coating. The cultures were used: 2D with Jurkat T cells (JTCs) on plastic surface; 2D co-culture of JTCs and AMMSCs on plastics; 3D with JTCs and CP matrix; 3D with JTCs, AMMSCs, and CP matrix. Findings & Conclusion: Both 2D and 3D JTC cultures showed an increase in the CD45RO receptor expression that led to increasing number of CD45RO+CD45RA+ cells. Probably, JTCs restored the partial maturation and differentiation. Vice versa, the expression of CD45RO receptor and the number of CD45RO+CD45RA+ cells decreased in case of JTCs and AMMSCs co-cultivation. A similar reaction of the cells was revealed in the 3D culture of JTCs and AMMSCs. Thus, JTCs with AMMSCs co-cultivation may

Modern biomaterial biocompatibility research focuses the biomaterial hierarchic effects on multipotent mesenchymal stromal cell (MMSC) behavior (that occur at the nano-, micro- and macroscales) because MMSCs are the fundamental units that produce/regenerate bone tissue. Leukemia initiation and progression are connected with a disfunction of health cell microenvironment and MMSCs. Continuous monitoring of MMSC and tumor cell interaction is a promising tool for oncology, cellular biology, biotechnology and environmental research. The aim was to investigate a modulation of in vitro interaction of human MMSCs and leukemic T lymphoblast-like cells (Jurkat T cells) caused by micro-arc multilevel calcium phosphate (CP) coating with the help of Cell-IQ and RTCA advanced tools for continuous monitoring. An average velocity of cell division (AVCD) of human adipose-derived MMSCs (hAMMSCs) contacted in vitro with allogenic Jurkat line of human leukemic T lymphoblasts (Jurkat T cells) was studied by means of Cell-IQ v2 MLF integrated phase-contrast microscopic platform for real-time surveillance imaging of living cells. Both 50 mu L suspensions (5x10(4) viable karyocytes) of the CD73CD90CD105(+) adherent cells and Jurkat T cells were applied into the center of the well of 12-well plastic plates for 7 days at 100% humidity in a 5% CO2 atmosphere at 37 degrees C until a monolayer formation. A nutrient medium was once replaced. To determine cell invasion (chemotactic motility) through 8 mu m pores the real-time cell analysis (RTCA DP Analyzer) with the CIM-plate was used. AVCD of fibroblast-like adherent hAMMSCs was 0.27-0.63 divisions/h. CP coating diminished significantly the percent of dividing hAMMSCs contacted with leukemic Jurkat T cells. RTCA system showed significant hAMMSC invasion towards tumor cells and not vice versa. For all this, cellular interaction led to increasing viability of Jurkat T cells and decreasing hAMMSC viability. Thus, tumor Jurkat T cells could control a fate of health hAMMSCs and promote stromal microenvironment for survivability of tumor clones by means of secretable molecular products. Multilevel micro-arc CP coating forms bone-like inorganic structure that is capable to modulate in vitro interaction of human MMSCs and leukemic T lymphoblasts. The results obtained may be useful for replacement surgery applications of orthopedic implants in cancer patients.

Using a multiplex kit the secretion of a number of cytokines, chemokines, and growth factors has been investigated in vitro in a culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) under conditions of their osteogenic differentiation caused by 14-day contact with a calcium phosphate (CP) surface of different roughness. Bilateral X-ray amorphous CP coatings were prepared on the samples of commercially pure titanium in the anodal regime using a microarc method. The electrolyte consisted of aqueous orthophosphoric acid (20 wt %), calcium carbonate (9 wt %), and synthetic hydroxyapatite nanopowder (6 wt %, particle diameter of 10–30 nm with single agglomerates up to 100 nm). hAMMSCs isolated from lipoaspirate were co-cultured after 4 passages with the CP-coated samples at a final concentration of 1.5 × 105 viable karyocytes per 1.5 mL of standard nutrition medium (without osteogenic stimulators) for 14 days (determination of the [CD45,34,14,20], CD73, CD90, and CD105 cell immunophenotype; analysis of secretory activity) and 21 days (alizarin red S cell culture staining) with medium replacement every 3–4 days. Under conditions of in vitro contact with rough CP coating hAMMSCs differentiated into osteoblasts synthesizing the mineralized bone matrix; this was accompanied by a 2−3-fold increase in the proportion of [CD45,34,14,20]+ hemopoietic cells. The following humoral factors of hemopoietic niches acted as the signal molecules escalating in vitro the hemopoietic base in 14 days of differentiating three-dimensional culture of hAMMSCs: leukemia inhibitory factor (LIF) and stem cell factor (SCF) cytokines in the case of the mean index of CP roughness Ra = 2.4–2.6 µm or stromal derived factor-1 (SDF-1α, CXCL12 chemokine) in the case of Ra = 3.1–4.4 µm.

Secretion of 21 cytokines, chemokines and growth factors (LIF, SCF, SDF-1a, SCGF-b, M-CSF, MCP-3, MIF, MIG, TRAIL, GRO-a; IL-1a, IL-2ra, IL-3, IL-12(p40), IL-16, IL-18, HGF, TNF-b, b-NGF, IFN-a2, CTACK) has been studied in vitro in the culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) in conditions of its osteogenic differentiation caused by 14-day contact with calcium phosphate (CP) surface with different roughness. Bilateral X-ray amorphous CP coatings were prepared on the samples of commercially pure titanium in the anodal regime using a micro-arc method. An aqueous solution prepared from 20 wt% phosphoric acid, 6 wt% dissolved hydrohyapatite nanopowder (particle diameter 10-30 nm with single agglomerates up to 100 nm), and 9 wt% dissolved calcium carbonate was used to obtain CP coating. hAMMSCs isolated from lipoaspirate were co-cultured after 4 passages with the CP-coated samples at final concentration of 1.5´105 viable karyocytes per 1.5 mL of standard nutrition medium (without osteogenic stimulators) for 14 days (a determination of [CD45,34,14,20], CD73, CD90 и CD105 cell immunophenotype; an analysis of secretory activity) and 21 days (alizarin red S staining of culture) with medium replacement every 3-4 days. Under conditions of in vitro contact with rough CP coating hAMMSCs differentiated into osteoblasts synthesizing the mineralized bone matrix; this was accompanied by 2-3-fold increasing ratio of [CD45,34,14,20]+ hemopoietic cells. The following humoral factors of hemopoietic niches acted as the signal molecules escalating in vitro the hemopoietic base in 14 days of differentiating three-dimensional culture of hAMMSCs: either leukemia inhibitory factor (LIF) and stem cell factor (SCF) cytokines under mean index of CP roughness Ra=2.4-2.6 mm or stromal derived factor-1 (SDF-1a, CXCL12 chemokine) under Ra=3.1-4.4 mm.

Migration, proliferation, and osteogenic differentiation of human adipose-derived (AD) multipotent mesenchymal stromal cells (MMSCs) during in vitro modeling of indirect contact with calcium phosphate (CP) or nanoparticles of synthetic hydroxyapatite (HA) have been studied. The results were registered with electrode (real-time cell analysis, RTCA) or visual (Cell-IQ) systems of long-term observation of cell cultures. Bulk specimens were use in a Cell-IQ® v2 MLF device as pure titanium substrates (10 × 10 × 1 mm3) covered by a CP relief (roughness index Ra = 2.4–4.4 μm) bilateral coating that was prepared by the micr-arc method from an aqueous solution of orthophosphoric acid (20 wt %), calcium carbonate (9 wt %), and synthetic HA (6 wt %). HA crystallites (1 mg/mL) were fabricated by mechanochemical synthesis and served as an irritant in RTCA investigation. The Cell-IQ system identified a 3.5- to 10-fold decrease in cell number at the interface with CP coatings with differing roughness during 14-day cell culturing. After 21 days, it was accompanied by a weak reduction of MMSC antigen expression (CD73, CD90, and CD105) as opposed to an increase in MMSC osteogenic differentiation and intercellular-matrix mineralization. In turn, HA nanodispersion reduced the speed of MMSC migration by 1.5 times (P < 0.001) during 25-h RTCA recording, which simulated cell invasion through the microporous membrane (8-μm diameter). Inhibition of migration and cell division with increased osteogenic differentiation of MMSCs has been suggested to be a possible effect of biodegradation products of synthetic CP materials.

В научных работах детально описана роль интерлейкина-8 в регуляции системы врожденного иммунитета, однако влияние данного хемокина на компоненты адаптивного иммунитета все еще требует изучения. Цель данного исследования оценить влияние интерлейкина-8 на дифференцировку активированных Т-лимфоцитов человека в условиях клеточного культивирования in vitro. Из цельной крови условно здоровых доноров выделили мононуклеарные клетки, которые использовали для позитивной селекции CD3+ клеток методом магнитной колоночной сепарации и на фоне TCR-активации культивировали in vitro с рекомбинантным интерлейкином-8 в разных концентрациях (0,01; 0,10; 1,00; 10,00 нг/мл). Влияние интерлейкина-8 на субпопуляции Т-лимфоцитов человека оценивали методом проточной цитофлуориметрии по изменению количества молекул, определяющих субпопуляции Т-лимфоцитов (CD4, CD45RA, CD197). Установлено, что интерлейкин-8 способен направлять дифференцировку CD4+ наивных (CD45RA+CD197+) Т-лимфоцитов в сторону эффекторных (CD45RA-CD197-) Т-клеток. Выявлено увеличение числа активированных CD4Т-лимфоцитов с фенотипом клеток эффекторной памяти на фоне снижения количества наивных Т-клеток и Т-клеток центральной памяти (CD45RA+CD197-). Таким образом, совместное культивирование интерлейкина-8 и активированных Т-лимфоцитов приводило к переходу CD3+CD4+ и CD3+CD4субпопуляций в более поздние стадии дифференцировки. Полученные данные указывают на участие интерлейкина-8 в процессах, связанных с созреванием активированных клеток адаптивного иммунитета.Scientific papers describe in detail the role of interleukin-8 (IL-8) in the regulation of the innate immune system. However, the effect of this chemokine on the components of the adaptive immune system requires further investigation. This study aimed to evaluate the effect of IL-8 on the differentiation of activated human T-lymphocytes under the conditions of in vitro cell culture. From the whole blood of practically healthy donors we isolated mononuclear cells which were used for positive magnetic-activated sorting of CD3+ cells that were then cultured in vitro with recombinant IL-8 in different concentrations (0.01; 0.10; 1.00; 10.00 ng/ml) accompanied by TCR activation. The effect of IL-8 on the subpopulation of human T-lymphocytes was assessed using flow cytometry by the changes in the number of molecules that determine the subpopulations of T-lymphocytes (CD4, CD45RA, and CD197). We found that IL-8 is able to cause differentiation of CD4+ T-lymphocytes from naive (CD45RA+CD197+) to effector memory (CD4+CD45RA-CD197-) T-cells. Further, we identified an increase in the number of activated CD4T-lymphocytes with the phenotype of effector memory cells against the background of decreasing number of naive and central memory T-cells (CD45RA+CD197-). Thus, co-culturing of IL-8 and activated T-lymphocytes resulted in the transition of CD3+CD4+ and CD3+CD4subpopulations to the later stages of differentiation. The findings indicate the involvement of IL-8 in the processes associated with maturation of activated adaptive immune system cells.