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ULTRASTRUCTURAL CHANGES IN THE SPINAL CORD OF RATS WITH EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS IN THE BACKGROUND OF MESENCHYMAL STEM CELLS AND INTERLEUKIN-10

Аuthors: V. V. Vaslovych, L. D. Pichkur, T. A. Malysheva, S. T. Akinola, S. A. Verbovska, O. K. Toporova, N. S. Shuvalova

Pages: 17–30

Abstract

         

Introduction. Human umbilical cord derived mesenchymal stem cells (UCd-MSC) are multipotent. They are able to migrate to the sites of central nervous system damage, contribute to the restoration of the number of endogenous oligodendrocytes, produce immunomodulatory effects and are able to synthesize antiinflammatory cytokines (IL-10).

Purpose. To investigate the effect of UCd-MSC and IL-10 on the processes of de- and remyelination of nerve fibers of the spinal cord of rats with experimental allergic encephalomyelitis at the ultrastructural level.

Materials and methods. The study was performed on 23 white female rats, weighting 210 g. The relapsing course of EAE was obtained by inserting Freund's adjuvant into the rat’s hind limb pads. Six groups of animals were formed. 18 rats from EAE were intravenously or suboccipitaly administered in different combinations of IL-10 (0.1 μg) and MSC no transfected (1 million) and MSC (1 million) transfected with the IL-10 gene. Electron microscopy was used to investigate the ultrastructural changes in the lumbar spinal cord of experimental animals at the 35th and 60th day of the experiment. Also, the ratio of the width of the myelin sheath (MS) to the diameter of the axial cylinder (AC) was determined on the semi-thin sections. Statistical analysis of the results was performed using non-parametric rank discriminant Kruskel–Wallis analysis followed by application of the U-Mann-Whitney criterion using STATISTICA 6.1 statistical software package.

Results. In animals of all experimental groups, the peak of EAE was observed for 19–20 days and developed a chronic remitting flow. In the comparison group (EAE), part of the oligodendrocytes already in the early stages are in a para apoptotic state, the degree of which increases over time. Reactive modified swollen forms of oligodendroglia, along with destructive, are more often represented in the study group III EAE + IL-10. Hypertrophic oligodendrocytes are more likely to be detected after the introduction of MSCs. Oligodendrocytes of group V (EAE + MSK transfected with IL-10 gene) for 60 days of study have signs of functional tension (swollen mitochondria with crysts fragmentation). In all experimental animal groups, with the exception of the comparison group, a reduced degree and demyelination area with a simultaneous partial reduction of the intralamilar and almost complete – periaxonal edema. In the study group III (EAE + IL), none of the terms of the study showed normalization of the structure of myelin membranes. Figures of remyelination of nerve fibers in all experimental groups were detected already on the 35th day of the study. The ratio of the thickness of the myelin sheath to the diameter of the axial cylinder is statistically significantly reduced relative to the comparison group in all treated groups except for the III (EAE + IL) for 35 days and III (EAE + IL) and V (EAE + MSC (T) at 60 days of the experiment.

Conclusions. MSCs help reduce interlamilar, periaxonal edema and demyelination in animals with EAE.Suboccipitaly introduced into CSF IL-10 does not produce statistically significant effects, however, to some extent, strengthens the effect of MSC.In all animal groups and in all terms, MSCs promote stimulation of remyelination processes.

Keywords: experimental allergic encephalomyelitis, mesenchymal stem cells, morphological changes, interleukin-10.

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