Vol 10, No 1 (2015)


Development of implantable cell-tissue-engineering designs of auxiliary liver for the treatment of liver failure

Onishchenko N.A., Gulay Y.S., Shagidulin M.Y., Nikolskaya A.O., Bashkina L.V.


The paper analyzes the achievements and prospects of creating implantable cell- and tissue-engineering designs (CEDs and TEDs) of auxiliary liver to treat liver failure. Emphasizes the need to maintenance long-term and steady function of implantable CEDs and TEDs at the treatment of liver failure, by forming in them de novo hepatospecific structures and transformation of these structures in the new centers of restorative regeneration of damaged liver. CEDs and TEDs acquire these properties due to inclusion in their designs small-differentiated cells: liverspecific cells (parenchymal and non-parenchymal), cells, committed in hepatoid direction and bone marrow cells, adherent to the biocompatible and biodegradable 3D-material, simulating the properties of the extracellular matrix The article analyzes the advantages, disadvantages and prospects for using the major groups of matrices materials (biological, synthetic,inclusive biopolymer and tissue-specific composite materials, obtained by liver decellularization). Indicates that the biopolymer materials occupy a preferred place among biodegradable scaffolds as have not only biocompatible, but also the properties of biostimulants. Since the production of the TEDs requires the provision of adequate stereotypical distribution of different types of cells in the matrix is paid great attention to the production of micro-scale, medium-scale and large-scale TEDs of auxiliary liver. However, points out that none of the problems of producing TEDs liver (choice of sources and technologies to produce small-differentiated cells, the selection matrix and technology of cell-sowing, the choice of assembly technology TEDs) can not be considered definitively settled
Genes & Cells. 2015;10(1):6-17
pages 6-17 views

Recovery options of reproductive function of cancer patients due to transplantation of cryopreserved ovarian tissue

Abakushina E.V., Otoi T., Kaprin A.D.


Cancer patients survive at increasing rates, but successful treatment in younger patients often leads to reduced fertility. If damage to reproductive organs from treatment is unavoidable, cryopreservation of ovarian tissue can protect fertility for young patients prior to treatment. Well known that the mammalian ovary contains a huge stock of resting оосуtes. The large store of these small follicles creates a potential source of oocytes for fertilization. To utilize the potential female gametes stored in ovaries, it will be important to safe ovarian tissue before oocytes undergo degeneration during treatment of cancer cryopreservation and transplantation of ovarian tissue are two emerging techniques for fertility preservation, especially in yang cancer patients If these technologies are to become widely accepted, they need to be safe, easy to perform and must obtain favorable result. Recent advances in cryobiology have made it possible to preserve ovarian tissue with relatively little loss of viability. If gonadal toxicity of chemo radiotherapy is unavoidable, physicians also should be knowledgeable about options for fertility preservation and offer patients a referral to a fertility specialist. The ability of having genetically related children is an important issue for patients surviving cancer.
Genes & Cells. 2015;10(1):18-27
pages 18-27 views

The role of EF-hand Са2+/Mg2+-binding tescalcin protein in cell proliferation and differentiation

Kolobynina K.G., Solovyeva V.V., Slepak V.Z., Rizvanov A.A.


EF-hand Са2+/Мд2+-binding proteins are involved in many important processes in the body. Identification and analysis of the EF-hand motifs in the genome led to the discovery of novel Ca2+-binding proteins, which are potentially useful for biomedical applications. One of such molecules is tescalcin - 24 kDa protein with one EF-hand motif. Tescalcin plays an important role in differentiation of hematopoietic cells by regulating the expression of Ets family transcription factors via PMA-induced ERK-pathway. At the molecular level, it was shown to interact with subunit 4 of signalosome COP9 and Na+/H+-exchanger. Recently a potential use of tescalcin for cancer diagnostics was demonstrated
Genes & Cells. 2015;10(1):28-34
pages 28-34 views

Prospects of the tissue engineering lung development with the methods of regenerative medicine (review)

Kuevda E.V., Gubareva E.A., Sotnichenko A.S., Gilevich I.V., Gumenyuk I.S., Macchiarini P.


Deficit of donor organs, the need for lifelong immunosuppressive therapy, even in the case of successful lung transplantation, high risk of death in the postoperative period are forced to look for the new ways to treat terminally ill patients, requiring organ transplants. In the last decade, much attention is given to methods of regenerative medicine to repair or replace the function of damaged tissues and organs. Creating functional lungs in the laboratory will hopefully solve the problem of donor organs shortage. The study of morphological properties of biological scaffolds, deeper understanding of stem cells and progenitor cells behavior led to the idea of using decellularization methods followed by recellularization with autologous cells for tissue engineered trachea, lungs, heart and kidney creation Recellularized solid organs can perform organ-specific function in vitro conditions, indicating the potential clinical use of these methods This review presents the current data about lung decellularization and recellularization methods to increase efficiency and improve the quality of the biological scaffold and discusses the main aspects of lung transplantation in animal models and perspectives of lung bioengineering
Genes & Cells. 2015;10(1):35-40
pages 35-40 views

Lekishvili Experimental and morphological study of the bone plastic materials for surgical treatment of ENT pathology

Ahmedov S.M., Musina L.A., Kocharyan E.Z., Ryabov A.Y., Lekishvili M.V.


To study the possibility of application different materials during plastic surgery of the middle ear structures with background of purulent infection experimental model with cranial bone defects replacement was utilized. As the object of the experiment 17 rabbits «Chinchilla» were used. All animals underwent surgery with formation of artificial defects in parietal bones (diameter of 7 mm) followed by simultaneous grafting. Following the implantation of plastic materials animals underwent standard antibacterial and local anti-inflammatory therapy For the plastic of parietal bone defects at rabbits were used and studied: autologous plates from the parietal bone and the iliac crest, auricle's autologous cartilage plates, demineralized bone implants (DBI), derived with technology of producing osteoplastic material «Perfoost» (CITO) from cortical long bones, rabbit's auricle autologous cartilage preserved in 0. 2% thymol solution, synthetic plastic material, comprising a hydroxyapatite and antibiotic - “Collapan-D” and osteoplastic composite material obtained by adhesion of the autologous chips from iliac crest and the parietal bone of rabbits with fibrin bond “Byo-Glu”. Timing of the experiment consisted of 15 and 30 days, 3 and 6 months By 3 months of the experiment organotypical callus were obtained in cases of application pure autologous tissues (plates from rabbit's parietal and the iliac crest and, as well, as auricle's autologous cartilage plates) Similar results were obtained in the cases with DBI “Perfoost” and allogenic rabbit's auricle cartilage preserved in 0. 2% thymol solution. In the defects filled with “Collapan-D” and combined osteoplastic material with fibrin glue «Byo-Glu» at 6 months of the experiment most part of the material was resorbed and partially replaced by newly formed bone alternating with areas of connective tissue and remnants of implants
Genes & Cells. 2015;10(1):41-47
pages 41-47 views

The human dermal fibroblasts cultivation on the porous polytetrafluoroethylene membrane for a scleroplastic

Blinova M.I., Yudintceva N.M., Vershevskaja E.A., Dghanaeva Z.N., Astakhov Y.S., Tomson V.V., Potokoin I.L., Galibin O.V., Pinaev G.P.


The cell technologies began to apply in ophthalmology actively. The aim of this investigation is to test the porous polytetraphtorethilen membrane as a substrate for the human dermal fibroblasts growth with further using for scleroplastik. It was shown by the scan electron microscopy that this membrane is a suitable substrate for a dermal fibroblast growth. Also it was shown, that the tissue-like structures are formed on the membrane previously seed with cells formerly than on membrane without cells in result of this membrane implantation into rabbit eye The obtained results suggest that dermal fibroblasts growing on porous polytetraphtorethilen membrane may be used in ophthalmology for the restoration of the damage or losed tissues. The clinical example mentioned in this work is evidence in this conclusion
Genes & Cells. 2015;10(1):48-54
pages 48-54 views

Nanodiamond composite scaffolds for human skin fibroblasts cultivation

Nashchekina Y.A., Margulis B.A., Gordeev S.K., Blinova M.I., Guzhova I.V.


Carbon materials are widely evaluated as scaffolds for cultured cells in “regenerative” bone surgery. In this research we investigated the interaction of human skin fibroblasts with diamond carbon composites scaffolds. Scaffolds with 10-100 nm pore diameter were obtained from the dispersed powders of diamond with particle size - from 2 nm to 5 microns It was shown that all scaffolds were not toxic for cultured cells. The highest number of cells adhered on the scaffolds with average pore size of 50-100 nm Scaffold with a pore size of 50 nm contribute to the fibroblasts parallel orientation of their surfaces
Genes & Cells. 2015;10(1):55-60
pages 55-60 views

Different expression of hematopoietic-supporting genes in cord, placental and bone marrow mesenchymal stromal cells

Kostjunina V.S., Petyovka N.V., Potapnev M.P.


Human multipotent mesenchymal stromal cells (MMSC) from bone marrow (BM), umbilical cord (UC) and chorion villi (CV) were isolated and cultured in xeno-free media supplemented with AB human serum There were no differences in expression of CD31, CD33, CD34, CD45, CD90, СD105, CD117, HLA-ABC, HLA-DR between BM, UC and CV MMSC Human AB serum (5%) accelerated proliferation of UC MMSC in vitro. Expression of genes opn, scf, cxcl12, il-3, il-6, il-8, il-11, g-csf, gm-csf, epo, and nes was studied in Real-Time PCR. Up-regulation the expression gene nes in CV MMSC and genes g-csf and il-11 (but 6-fold down-regulation of cxcl12) in UC MMSC, was revealed when compared to BM MMSC (p<0. 03). Expression of il-6 and epo genes in MMCS did not differ. Trend to 10-fold increased expression of gene il-8 as well as cytokine content in UC MMSC vs BM MMCS was demonstrated CV MMSC revealed more differences from the UC MMSC than from the BM MMSC in expression of hematopoietic-supporting genes (opn, scf, il-11, p<0.03). Various abilities of mesenchymal cells from human BM, UC and placenta to express hematopoietic-supporting genes had to be taken into account in application of MMCS for generation of hematopoietic stem cells in vitro
Genes & Cells. 2015;10(1):61-68
pages 61-68 views

Plasticity of bone marrow-derived stromal cells at grafting onto neural tissue after ischemic injury in vitro

Rybachuk O.A., Kyryk V.M., Poberezhnyi P.A., Butenko G.M., Pivneva T.A.


Bone marrow-derived multipotent mesenchymal stromal cells (BM-MMSCs) are able to confer beneficial effects after transplantation into neural tissue with ischemic injury. This effect is probably caused by the release of trophic factors, although the possibilities of replacement of dead neural cells by BM-MMSCs are not excluded. The aim of this study was to identify the ability of BM-MMScs to differentiate into cells of the nervous tissues and their neuroprotective effect in direct contact with nervous tissue damaged by ischemia Therefore, we investigated this interaction by in vitro model of organotypic hippocampal tissue to avoid affecting the immunological processes in the conditions after transplantation in vivo. Ischemic injury induced by oxygen-glucose deprivation The potential of differentiation of transplanted multipotent mesenchymal bone marrow stromal cells to neural direction was assessed for 14 days after the ischemic injury. At the 7 th day after the oxygen-glucose deprivation and transplantation the multipotent mesenchymal bone marrow stromal cells differentiated into microglial cells, and on the 14th day - as in microglial cells and in mature oligodendrocytes These findings suggest that the transplanted stem cells respond to signals from the microenvironment of the injured tissue of the recipient, which in turn may trigger and regulate cell differentiation as well as to determine the direction of migration
Genes & Cells. 2015;10(1):72-82
pages 72-82 views

Morphofunctional reaction of T-lymphoblasts of Jurkat line on the titanium oxides coating

Khlusov I.A., Litvinova L.S., Shupletsova V.V., Sokhonevich N.A., Khaziakhmatova O.G., Khlusova M.Y., Sharkeev Y.P., Pichugin V.F.


Human leukemic T-lymphoblastoid cells (hereinafter Jurkat T-cells) which was applied actively for modeling of T-lymphocytes reaction have been used in vitro We have examined morphofunctional response of Jurkat T-cells to 24-h in vitro contact with titanium substrates (12x12x1 mm3) covered by titanium oxide (TiO, TiO2) bilateral coating that was prepared by microarc method from an aqueous solution of 20 mass % orthophosphoric acid. 27-98% of immortalized cells in control culture (without an addition of the specimens) had CD3+CD4+CD71+CD45RA+ immunophenotype and secreted IL-2, IL-4, IL-8, IL-10 and TNFα, but not IL-1b and IL-6. Other markers of cell activation, differentiation, maturation and death (CD8, CD16, CD56, CD25, CD95) were found at 0 - 2. 5% of the cell population. The addition of bulk specimens with oxide coating reduced statistically CD3, CD4, CD8 и CD95 membrane markers presentation on Jurkat T-cells and decreased IL-4 and TNFα secretion Structural (antigens expression) and functional (cytokines secretion) Jurkat T-cells inactivation was not connected with the generation of intracellular reactive oxygen species (ROS), and was not mediated by TiO2 nanoparticles (diameter of 14±8 nm; doses of 1 mg/L or 10 mg/L) which can be released under samples biodegradation. Spearman's correlation analysis showed the inhibiting action of the oxide surface roughness in the range of Ra = 2,2-3,7 μm on the number of viable Jurkat T-cells (rs = -0. 95; n = 9; p<0. 0001) due to an elevating portion of necrotic forms in the cellular population, mainly In turn, magnitude of negative electrostatic potential of the oxide surface rose linearly (r = 0. 6; p<0. 000001, n = 60) with increase in the Ra roughness index The roughness of the oxide coating induces its surface voltage that seems to promote morphofunctional suppression of tumor immune cells by electrostatic/biological mechanisms are not connected with intracellular ROS generation
Genes & Cells. 2015;10(1):83-90
pages 83-90 views

Modification of polycaprolactone scaffolds with vascular endothelial growth factors for potential application in development of tissue engineered vascular grafts

Sevostyanova V.V., Golovkin A.S., Antonova L.V., Glushkova T.V., Barbarash O.L., Barbarash L.S.


In this study, we investigated a biological activity of nonwoven polycaprolactone scaffolds for controlled delivery of vascular endothelial growth factors. The tube scaffolds with incorporated vascular endothelial growth factors were fabricated by method of electrospinning. The polycaprolactone scaffold containing growth factor provided a morphology similar to the native extracellular matrix. The sustained release of biologically active growth factor from scaffold was observed for 80 days The assessment of adhesion and proliferation of multipotent mesenchymal stem cells and endothelial cells on the material surface showed that scaffolds with vascular endothelial growth factors are able to maintain the cellular activity. Results of study demonstrated that incorporated growth factors provide active proliferation of endothelial cells on porous material and cells penetration inside the scaffold. This approach to the creation of a biologically active environment in the scaffold has a great potential in the development of grafts for blood vessels regeneration
Genes & Cells. 2015;10(1):91-97
pages 91-97 views

Stimulation of rat's sciatic nerve regeneration by transplantation of human adipose derived multipotent mesenchymal stromal cells

Masgutov R.F., Masgutova G.A., Salafutdinov I.I., Shulman A.A., Zhuravleva M.N., Gallyamov A.R., Bogov A.A., Bogov A.A., Rizvanov A.A.


Peripheral nerve injury is a potential threat to the loss of function of an extremity. Despite the ability of neurons to axonal regeneration, nerve injury causes a significant level of neuronal death in spinal ganglia L4-L5, that leads to incomplete functional recovery. The influence of xenotransplantation of human adipose derived multipotent mesenchymal stromal cells on the process of regeneration of rat's sciatic nerve. It is shown that cell therapy contributes to a significant neuronal survival in spinal ganglia L5 after autonerve grafting as well as restore microcirculation and improves recovery of motor activity
Genes & Cells. 2015;10(1):98-102
pages 98-102 views

Influence of allogeneic hematopoietic stem/ progenitor cells treated with IL-3 on structural-functional heart changes under experimental postinfarction heart failure in long-term period of observation

Severova E.A., Berkinbayev S.F., Nugmanova M.N., Pominova N.M., Perfilyeva Y.V., Supniyazova T.A., Denisov Y.D., Belyaev N.N.


The aim of this research was to study the influence of allogeneic hematopoietic stem/ progenitor cells infused intravenously to rats in acute period of izoproterenol-induced myocardial infarction on structural and functional changes of the heart in long-term period of observation (over 1 year) The postinfarction heart failure was created by intra-abdominal injection of isoproterenol solution Allogenic CD117+ hematopoietic stem/ progenitor cells were extracted from bone marrow by immunomagnetic separation and were induced for CXCR4 expression and TGFb production by cultivation for three days in presence of interleukine-3. Obtained cells were injected intravenously to rats on 9 day after isoproterenol administration. One year later structure and function of the myocardium were investigated by epicardial echocardiography in M-modal mode with the apparatus «VIVID-3» supplied by 3. 5 MHz transduser The diastolic function was examined by Doppler echocardiography method in the impulse mode One year later, in the group of infarcted animals with hematopoietic stem/ progenitor stem cell injection the clinically and statistically significant reduction of the left ventricle weight and relative thickness of the left ventricle wall were revealed; furthermore - the reduction of the left ventricle back wall thickness, interventricular septum thickness and aorta diameter were observed in comparison with the group without treatment Doppler echocardiographic test has demonstrated an improvement of cardiac diastolic function in the group of infarcted animals with hematopoietic stem/ progenitor cell injection: statistically significant reduction of proportion of the animals with hypertrophic and restrictive mode of transmitral blood flow Obtained data have shown that single intravenous injection of hematopoietic stem/ progenitor cells treated with IL-3 ex vivo reduces a risk of serious cardiovascular complications after myocardial infarction at long-term period of observation in rats Proposed approach may be useful in improving of cell therapy effectiveness for heart failure treatment
Genes & Cells. 2015;10(1):103-110
pages 103-110 views

Nucleic acid-protein identification: novel protein classification based on nucleic acid-protein interaction

Krylov A.S., Zhdanov R.I.


Several basic properties of proteins are used for their classification. Some proteins can bind DNA and RNA in a specific way. We propose a simple method for classification of nucleic acids binding proteins which we call «nucleic acid-protein fingerprint» The method is based on usage of short oligonucleotides, immobilized in hydrogel for hybridization with proteins. First, we have found the shortest oligonucleotide capable for specific binding with nucleic acids. Experiments with samples from 2 to 12 bases length have shown that tetramers with 2 chains of 5-nitroindole (universal base) and 2 chains of normal bases can recognize proteins. Thus, we have made a smallest universal protein recognizing biochip containing 16 dinucleotides. Physical length of them is 4 chains, two chains are from 5-nitroindole and two chains are normal DNA bases We manufactured that biochip and investigated binding of dye labeled proteins with them. The hybridization patterns of two proteins RNase A and Binase have shown remarkable difference specificity and difference between proteins Biochip of this type with three of four remarkable proteins will have much more specificity. Thus, we proposed and experimentally confirmed a new type of proteins classification
Genes & Cells. 2015;10(1):111-114
pages 111-114 views

Current methods in experimental angiogenesis investigation

Livanova A.A., Deev R.V., Rizvanov A.A.


Growing interest in angiogenesis, a key component in the development of different diseases, requires the use of a suitable experimental model to simulate neovascularization in a laboratory. In recent years, with the development of novel therapeutic strategies, based on angiogenesis regulation, this problem has become especially important. Current in vitro and in vivo models are characterized with a variety of disadvantages, which impede results interpretation. Thus, in vitro assays provide estimation of discrete endothelial cells characteristics, which alter from the same ones in the native microenvironment. The use of in vivo assays is accompanied with difficulties in testing agent delivery and quantitative analysis of its angiogenic activity In view of these complications, the use of a combination of assays is recommended while planning the experiment in this area. The aim of this review is to critically analyze angiogenesis assays, currently used to perform fundamental investigation as well as preclinical tests of developing therapeutic agents
Genes & Cells. 2015;10(1):115-127
pages 115-127 views

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