Republic Clinical Hospital

Kazan, Russia

Republic Clinical Hospital

Kazan, Russia
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Mukhamedshina Y.O.,Kazan State Medical University | Gilazieva Z.E.,Kazan Federal University | Arkhipova S.S.,Kazan Federal University | Galieva L.R.,Kazan Federal University | And 6 more authors.
Neural Plasticity | Year: 2017

In this study, we examined the efficacy of human umbilical cord blood mononuclear cells (hUCB-MCs), genetically modified with the VEGF and GDNF genes using adenoviral vectors, on posttraumatic regeneration after transplantation into the site of spinal cord injury (SCI) in rats. Thirty days after SCI, followed by transplantation of nontransduced hUCB-MCs, we observed an improvement in H (latency period, LP) and M(Amax) waves, compared to the group without therapy after SCI. For genetically modified hUCB-MCs, there was improvement in Amax of M wave and LP of both the M and H waves. The ratio between Amax of the H and M waves (Hmax/Mmax) demonstrated that transplantation into the area of SCI of genetically modified hUCB-MCs was more effective than nontransduced hUCB-MCs. Spared tissue and myelinated fibers were increased at day 30 after SCI and transplantation of hUCB-MCs in the lateral and ventral funiculi 2.5 mm from the lesion epicenter. Transplantation of hUCB-MCs genetically modified with the VEGF and GNDF genes significantly increased the number of spared myelinated fibers (22-fold, P>0.01) in the main corticospinal tract compared to the nontransduced ones. HNA+ cells with the morphology of phagocytes and microglia-like cells were found as compact clusters or cell bridges within the traumatic cavities that were lined by GFAP+ host astrocytes. Our results show that hUCB-MCs transplanted into the site of SCI improved regeneration and that hUCB-MCs genetically modified with the VEGF and GNDF genes were more effective than nontransduced hUCB-MCs. © 2017 Y. O. Mukhamedshina et al.

Masgutov R.,Kazan Federal University | Chekunov M.,Republic Clinical Hospital | Zhuravleva M.,Kazan Federal University | Masgutova G.,Kazan Federal University | And 5 more authors.
BioNanoScience | Year: 2017

This paper presents a clinical case of successful ulnar pseudarthrosis treatment using a gene-activated bone allograft containing VEGF (vascular endothelial growth factor) and BMP2 (bone morphogenetic protein 2) in the form of a multicystron plasmid. Demineralized bone matrix with applied recombinant plasmid DNA was grafted into the bone defect using the classical open surgical approach. Two months after the surgery, the patient noticed the disappearance of pain including pain during activity. On X-rays of ulna, signs of union in the form of callus formation were found. © 2016, Springer Science+Business Media New York.

Masgutova G.,Kazan Federal University | Mukhamedshina Y.,Kazan Federal University | Sergeev M.,Kazan State Academy of Veterinary Medicine | Shulman I.,Republic Clinical Hospital | And 3 more authors.
BioNanoScience | Year: 2017

Mesenchymal stem cells (MSCs) are considered the most versatile cells for cell therapy—particularly for repair of injuries to the central nervous system. Recently, the use of dental pulp MSCs (DP-MSCs) for spinal cord regeneration has became especially important. We describe a surgical procedure for extracting pig teeth to obtain DP-MSCs using protocols for direct and enzymatic isolation of DP-MSCs followed by cultivation. Our study shows that primary pulp cultures of DP-MSCs are established 5 days after enzymatic digestion and 7–10 days following attachment of the digested minced tissue to the bottom of a plate. Though in the first few days the rate of primary expansion for cultures generated by direct isolation was lower than the rate of enzymatic digestion, this difference leveled off on days 14–18 of culture. For DP-MSCs isolation, we recommend the use of deciduous succedaneous lateral incisors and canine teeth of pigs as well as deciduous premolars from the first dentition of young pigs (up to 3 months). © 2016, Springer Science+Business Media New York.

Rizvanov A.A.,Yeditepe University | Rizvanov A.A.,Republic Clinical Hospital | Guseva D.S.,Hannover Medical School | Guseva D.S.,Kazan State Medical University | And 11 more authors.
Experimental Biology and Medicine | Year: 2011

Current therapy of a number of neuropsychiatric maladies has only symptomatic modality. Effective treatment of these neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), may benefit from combined gene/stem-cell approaches. In this report, mononuclear fraction of human umbilical cord blood cells (hUCBCs) were transfected by electroporation with dual plasmid constructs, simultaneously expressing vascular endothelial growth factor 165 (VEGF165) and human fibroblast growth factor 2 (FGF2) (pBud-VEGF-FGF2). These genetically modified hUCBCs were injected retro-orbitally into presymptomatic ALS transgenic animal models ( G93A mice). Lumbar spinal cords of rodents were processed for immunofluoresent staining with antibodies against human nuclear antigen (HNA), oligodendrocyte-specific protein, S100, iba1, neuronal β3-tubulin and CD34. Co-localization of HNA and S100 was found in the spinal cord of mice after transplantation of genetically modified hUCBCs over-expressing VEGF-FGF2. Double staining in control animals treated with unmodified hUCBCs, however, revealed HNA+ cells expressing iba1 and CD34. Neuron-specific β3-tubulin or oligodendrocyte-specific protein were not expressed in hUCBCs in either control or experimental mice. These results demonstrate that genetically naïve hUCBCs may differentiate into endothelial (CD34+) and microglial (iba1+) cells; however when over-expressing VEGF-FGF2, hUCBCs transform into astrocytes (S100+). Autocrine regulation of VEGF and FGF2 on hUCBCs, signal molecules from dying motor neurons in spinal cord, as well as self-differentiating potential may provide a unique microenvironment for the transformation of hUCBCs into astrocytes that eventually serve as a source of growth factors to enhance the survive potential of surrounding cells in the diseased regions. Copyright © 2011 by the Society for Experimental Biology and Medicine.

Yergeshov A.A.,Kazan Federal University | Siraeva Z.Y.,Kazan State Medical University | Kazakova R.R.,Kazan Federal University | Mullin R.I.,Republic Clinical Hospital | And 7 more authors.
Genes and Cells | Year: 2015

We prepared an experimental sample of gelatin based cryogel membrane with pore size of 50-150 μm. Confocal microscopy and LDH assay showed that the cryogel macroporous structure promotes migration and proliferation of human skin fibroblasts within the matrix in vitro. To assess in vivo effect of the cryogel an excision wound model in rats was tested. The cryogel significantly increases the number of fibroblasts as well as the density and order of produced collagen fibers in the dermis to day 7 of the wound healing process. The results suggest the stimulating effect of the gelatin cryogel on fibroblasts activity and demonstrate its potential for skin regeneration.

Masgutov R.F.,Kazan Federal University | Masgutova G.A.,Kazan Federal University | Rogojin A.A.,Kazan State Medical University | Zhuravleva M.N.,Kazan Federal University | And 7 more authors.
Genes and Cells | Year: 2015

The effectiveness of application of biodegradable conduit (NeuraGen; INTEGRA, USA) in combination with adipose derived multipotent mesenchymal stem cells and fibrin sealant (Tissucol-Kit; Baxter AG, Austria) on posttraumatic peripheral nerve regeneration was investigated using a rat model of sciatic nerve injury. We demonstrate that our proposed cell therapy confers a neuroprotective effect under our experimental design.

Mukhamedshina Y.O.,Kazan Federal University | Masgutov R.F.,Kazan Federal University | Masgutova G.A.,Kazan Federal University | Zuravleva M.N.,Kazan Federal University | And 5 more authors.
Genes and Cells | Year: 2015

In this study we used promising therapeutic strategy for stimulation of peripheral nerve regeneration - implantation of poly(ε-caprolactone) nerve conduit filled with fibrin hydrogel Tissucol in combination with mesenchymal stem cells. In vitro studies showed that the coating by Tissucol of poly(ε-caprolactone) substrate promotes cell proliferation. In vivo results of peripheral nerve defect reconstitution in rats using the conduit based on poly (ε-caprolactone) filled with fibrin hydrogel with mesenchymal stem cells confirmed the effectiveness of the proposed approach.

Rocha N.P.,Federal University of Minas Gerais | Teixeira A.L.,Federal University of Minas Gerais | Coelho F.M.,Federal University of Minas Gerais | Caramelli P.,Federal University of Minas Gerais | And 11 more authors.
Molecular and Cellular Neuroscience | Year: 2012

Objectives: Among several other factors, the neuro-toxic β-amyloid peptide (βAP)-induced inflammatory mechanisms have also been implicated in the pathogenesis of Alzheimer's dementia (AD). Cytokines have recently emerged as prime candidates underlying this immune reaction. The purpose of this study was to evaluate the inflammatory response of peripheral blood mono-nuclear cells (PBMC) in AD. Design: Cross-sectional (observational) study. Setting: Behavioral and cognitive neurology clinic of the Universidade Federal de Minas Gerais in Belo Horizonte, Brazil. Participants: AD patients (n = 19), healthy elderly (n = 19) and young (n = 14) individuals. Measurements: Cytokine levels were assessed by enzyme-linked immuno-sorbent assay (ELISA) after exposing cells to a broad range of βAP concentrations (10 -4-10 -10M) as a stimulus. AD samples were weighed against leukocytes harvested from non-demented young and elderly subjects. Results: Cytokine production of PBMCs in the youth was characterized by low baseline levels when compared to cells from the older generation. In the aging population, AD cells were distinguished from the healthy elderly sub-group by an even higher basal cytokine secretion. The low resting concentration in young individuals was markedly increased after treatment with βAP, however cells from the elderly, irrespective of their disease status, showed unchanged cytokine release following βAP administration. Non-specific activation of PBMCs with anti-CD3/CD28 antibodies resulted in elevated interleukin (IL)-1β concentrations in AD. Conclusions: These results demonstrate a general over-production of cytokines and resistance to βAP in the old comparison group, with a more pronounced disruption/boosted pattern in AD. Our findings are in line with the hypothesis of "inflammaging", i.e. an enhanced inflammatory profile with normal aging and a further perturbed environment in AD. The observed cytokine profiles may serve as diagnostic biomarkers in dementia. © 2011 Elsevier Inc.

Martins L.C.A.,Federal University of Minas Gerais | Rocha N.P.,Federal University of Minas Gerais | Torres K.C.L.,Federal University of Minas Gerais | dos Santos R.R.,Federal University of Minas Gerais | And 15 more authors.
Journal of Neuroimmunology | Year: 2012

Alzheimer's dementia (AD) is a degenerative brain disorder characterized mainly by cholinergic failure, but other neuro-transmitters are also deficient especially at late stages of the disease. Misfolded β-amyloid peptide has been identified as a causative agent, however inflammatory changes also play a pivotal role. Even though the most prominent pathology is seen in the cognitive functions, specific abnormalities of the central nervous system (CNS) are also reflected in the periphery, particularly in the immune responses of the body. The aim of this study was to characterize the dopaminergic and serotonergic systems in AD, which are also markedly disrupted along with the hallmark acetyl-choline dysfunction. Peripheral blood mono-nuclear cells (PBMCs) from demented patients were judged against comparison groups including individuals with late-onset depression (LOD), as well as non-demented and non-depressed subjects. Cellular sub-populations were evaluated by mono-clonal antibodies against various cell surface receptors: CD4/CD8 (T-lymphocytes), CD19 (B-lymphocytes), CD14 (monocytes), and CD56 (natural-killer (NK)-cells). The expressions of dopamine D3 and D4, as well as serotonin 5-HT1A, 5-HT2A, 5-HT2B and 5-HT2C were also assessed. There were no significant differences among the study groups with respect to the frequency of the cellular sub-types, however a unique profound increase in 5-HT2C receptor exclusively in NK-cells was observed in AD. The disease-specific expression of 5-HT2C, as well as the NK-cell cyto-toxicity, has been linked with cognitive derangement in dementia. These changes not only corroborate the existence of bi-directional communication between the immune system and the CNS, but also elucidate the role of inflammatory activity in AD pathology, and may serve as potential biomarkers for less invasive and early diagnostic purposes as well. © 2012 Elsevier B.V.

Chuenkova E.A.,Republic Clinical Hospital | Simushkin S.V.,Kazan Federal University | Ziganshina L.E.,Kazan Federal University
Eksperimental'naya i Klinicheskaya Farmakologiya | Year: 2016

Analysis of the results of pharmacological phenotyping using antipyrine test prior to providing anesthesia for laparoscopic cholecystectomy showed that trimeperidine (promedol) dosing with allowance for the total oxidative capacity of liver and the patient mass allows the periods of post- Anesthetic rehabilitation to be controlled. Clear algorithm of trimeperidine dosing based on established indices of the total oxidative capacity of liver and is yet nor developed because of restricted sampling set. The obtained results show expediency of using and studying antipyrine test as a simple, cheap, and informative method of individual anesthesia dosing for increasing the adequacy of general anesthesia.

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