Dysferlinopathies is a group of autosomal-recessive inherited neuromuscular diseases, which are characterized by defect in mRNA expression or in functionioning of dysferlin protein, appearing in about 1/200 000 births. Dysferlin is encoded by DYSF gene (Dystrophy-associated fer-1-like). It's disruption can cause various types of primary dysferlinopathies, which include Miyoshi myopathy (MM), Limb-girdle Muscular Dystrophy type 2B (LGMD2B) and distal myopathy with anterior tibial onset. Also, dysferlin deficiency can be associated with other diseases, such as caveolin- and calpainopathies. Here we discuss dysferlin protein structure and function, it's clinical phenotypes, known animal models and developing treatment strategies for dysferlinopathies.
The destructive forms of periodontal disease have a high prevalence. Our study was aimed to evaluate the effectiveness of tissue-engineering osteoplastic material in treatment of patients with periodontal pathology. We have obtained autologous gingival fibroblasts of patients, combined with the hydroxyapatite carriage and transplanted on a standard surgical treatment of periodontitis destructive forms. As a control we used the material without cells implanted into periodontal defects of other affected teeth of the same patients. We have shown the better effect of tissue-engineering osteoplastic material regarding the periodontal pockets depth, but significant difference in bone regeneration and other indicators has not been revealed. Thus, a continuation of researches is required to modify the technology of tissueengineering osteoplastic material creation and to choice the more optimal carrier for gingival fibroblasts.
Development of new effective osteoplastic materials is highly requested in practice of traumatology and orthopedics, oral and maxillofacial surgery. The goals of our research were design and construction of gene-activated bone graft (GABG) consisting of collagen/hydroxyapatite scaffold and plasmid DNA encoding vegf-a165 and evaluation of its biological effect in vitro and in vivo. We have shown that GABG co-incubation with multipotent mesenchymal stromal cells increased their VEGF protein expression. After GABG implantation into parietal bones defects the transfection of «recipient bed cells» was observed and accompanied by more pronounced angiogenesis as compared with control. The lager volume of bone regenerate was in case of GABG on 15 and 30 days after application. The source of reparative osteogenesis was not only parietal bones but also the GABG fragments (even from central part of the defect) majority of which were surrounded by newly formed bone tissue. In control group no osteoinductive effect has been observed. Thus, GABG with plasmid DNA encoding VEGF-A165 possesses angiogenic activity providing osteoinductive properties.
Development of bioengineered scaffolds of internal organs is one of the priority areas of tissue engineering. Decellularization allows to obtain biological (natural) scaffolds while preserving extracellular matrix and three-dimensional structure of organs. The primary goals of the present research were to investigate pathological characteristics of the decellularized rat heart scaffold and evaluate adhesion and viability of multipotent mesenchymal stromal cells (MMSC) during recellularization. Rat hearts were decellularized using a modified detergent-enzymatic method including sodium deoxycholate and DNAse. The results of morphological studies have confirmed the safety of extracellular matrix proteins and patency of the coronary vessels. Mechanical testing of decellularized and native samples of rat heart showed an increase of mechanical properties of the matrix during decellularization. During the conducted experiments on recellularization of obtained scaffold has been shown that the extracellular matrix was not toxic for MMSC which were viable and maintained their metabolic activity during prolonged cultivation on the scaffold.
It is known that human cord blood hematopoietic stem cells (HSC) are able to differentiate into hepatocytes. This ability can be widely used in treatment of various liver diseases. However, there are some genetic diseases of liver, when the application of autologous stem cells is not possible. So it could be very helpful to develop methods of genetic modification of stem/progenitor cells. However, it should be proved that genetic modification does not change the properties of HSC. We performed partial hepatectomy for the white mongrel male rats and injected human umbilical cord blood mononuclear cells transfected by gene of green fluorescent protein (GFP) into the spleen. Paraffin sections of the liver were stained with antibodies to stem cell factor receptor, human leukocyte antigen, a-smooth muscle actin, enhanced GFP, cytokeratin 19, hepatocyte specific antigen, human a-fetoprotein. Also we used a double-immunohistochemical staining to detect expression of stem cell factor receptor and desmin, enhanced GFP and cytokeratin 19. Our study showed that human cord blood mononuclear cells transfected by gfp transplanted into the spleen of rats after partial hepatectomy migrated to the liver and acquired the phenotype of hepatocytes, cholangiocytes and sinusoidal cells. At the same time the differentiation of such transplanted cells into myofibroblasts, as it was previously shown, does not occur. Hepatoblasts and hepatocytes found in the liver of rats after transplantation of genetically modified and native cells express human hepatocyte specific antigen and a-fetoprotein that means they are functionally active.
Ability of mammalian liver to regenerate is one of the favorite examples of “regenerative medicine”. At the same time liver regeneration can not be viewed as a simple hypertrophy, it must have some appropriate steps. Understanding of these processes is crucial for correct interpretation of liver therapy results, especially after cellular therapy. But, unfortunately, original and first-hand data regarding changes in liver microstructure during regeneration is relatively scarce. This work was dedicated to study changes of liver microstructure during liver regeneration after partial hepatectomy in rats. We analyzed proliferative processes, perisinusoidal cells involvement, sizes of classical hepatic lobules, participation of bile ducts, branches of afferent and efferent hepatic vessels in liver regeneration on 1, 2, 3, 5, 7 postoperative days. Our results have shown that liver microstructure during regeneration after partial hepatectomy undergoes two stages: hypertrophy of hepatic lobules by proliferation of liver cells until 4th day and division of hepatic lobules by branching of bile ducts, hepatic artery, portal and central veins from 4th until 7th postoperative day.
Gene-cell therapy is a new step for the treatment of different human disorders including central nervous system degenerative diseases. In this review we focused on the last challenges in the field of human umbilical cord blood mononuclear cells transplantation - an attempt to support neuronal cells survival and to stimulate the neuroregeneration. As a potential therapy for the treatment of neurodegenerative diseases we reviewed the latest advances in gene modification of human umbilical cord blood mononuclear cells as a novel tool for the effective delivery of neuroprotective factors and growth factors in the injured or degenerative areas of the central nervous system under pathological conditions. The main topic of this review is the potential therapy of the amyotrophic lateral sclerosis - the progressive neurodegenerative disorder affecting primarily upper and lower motoneurons - by using genetically modified human umbilical cord blood mononuclear cells. The results from the up-to-date experiments indicated the opportunity to obtain differentiated macrophages, endothelial cells, or astrocytes from the genetically modified human umbilical cord blood mononuclear cells after their transplantation in the mouse model of the amyotrophic lateral sclerosis. Taken together, these data build the high-capacity platform for the supporting of degenerating neurons, structural and functional recovery of the brain and spinal cord after trauma, ischemia and other neurodegenerative disorders.
Nowadays there are findings that C-kit-positive and desmin-positive stellate cells help to regenerate endocrine part of pancreas. But still it is unknown about their role and the way of differentiation of these cells during pancreas regeneration. That's why the aim of our work was to study the dynamic of these cells population during alloxan diabetes in rats. The work was made on 33 rats with experimental diabetes. Blood glucose, insulin and glucagon levels were analyzed. Also the expression of desmin (marker of stellate cells), a-SMA (marker of myofibroblast), C-kit (marker of endocrine stem cells), insulin and glucagon (marker of differentiated a- and р-cells of Langerhans islets) was studied. The expression of desmin was found after one day of experimental diabetes in islets cells of pancreas. Maximum of these cells was after the third day of the experiment. Also after one day of the experiment C-kit-positive cells, which expressed insulin and glucagon were found. We suppose that stellate cells are the main factor of microenvironment for differentiation of C-kit-positive progenitor cells into р-cells through the stage of glucagon-positive cells because stellate pancreas cells can produce different growth factors and components of intercellular matrics.
The article presents a case report of successful treatment of pseudarthrosis of the femur with the use of autologous cells of the stromal vascular fraction (SVF) derived from adipose tissue. Autologous SVF cells mixed with fibrin glue were transplanted into bone defect by injection under the control of electro-optical converter. Two months after injection, the patient reported the disappearance of pain. After 4 months we observed the recovery of function of the knee, X-ray shows signs of the fracture healing through the formation of bone callus.
Today there is no effective approach to treat liver fibrosis and the only way is transplantation of donors' liver. Investigation of molecular-cellular mechanisms of liver fibrosis can help to discover new ways of slowing down or even reverse the process of fibrogenesis in liver. For a long time hepatic stellate cells were undeservingly blamed for being the major causer of liver fibrosis, because they were considered as the main source of myofibroblasts, that synthesize connective tissue extracellular matrix. In this particular work explantation approach was used to isolate cell culture from portal tracts. It was shown that received cells are portal fibroblasts, and, just as hepatic stellate cells, in case of liver alteration can differentiate in myofibroblasts, that express а-smooth muscle actin. During long-term cultivation it was shown that portal myofibroblasts can differentiate into fibroblasts and back on late passages. So, we can conclude, that there is a potency to reverse the process of fibrogenesis in liver.
In this article we describe a clinical case of successful treatment of neurotrophic ulcers of heel region using classical surgical techniques such as debridement and autodermoplastics combined with vacuum therapy and injection of double cassette expression plasmid encoding cDNAs of vascular endothelial growth factor and fibroblast growth factor 2. The outcome of therapy opens new horizons in the treatment of such diseases in short period of time.
On the model of rat spinal cord dosed contusion at Th8 level studied the effect of delivery into the area of damage of glialcell-line-derived neurotrophic factor gene using adenovirus-transduced of human umbilical cord blood mononuclear cells on motor recovery and maintaining a population of glial cells. The results show that the proposed method of gene-cell therapy can effectively stimulate the regeneration of posttraumatic spinal cord injury, which is manifested in the form of improved indicators of recovery of motor function, increasing the number of reactive astrocytes and oligodendrocytes progenitors.