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Adjuvant stem cell-based therapy in acute retinal injury after sodium iodate administration in mice: Morphological and functional study

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Machalinska A., Roginska D., Pius E., Stepniewski J., Kawa M., Jozkowicz A., Dulak J., Machalinski B.
Acta Neurobiologiae Experimentalis
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2013
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tom 73
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nr 1
Ophthalmic diseases, especially retinal degeneration belong to prominent causes of disability in developed countries. Thus, special attention has been focused on research aimed on establishing new protocols of efficient stem cell (SC)-based therapy of these disorders. The aim of this study was to determine and optimize a new strategy of SC-based therapy of selectively damaged retina after sodium iodate (NaIO3) administration in C57BL/6J mice. First, we sought to assess the regenerative mechanisms triggered after acute chemical injury of retinal pigment epithelium and neurosensory retina induced by NaIO3, in mice. The intravenous injection of NaIO3 provides a useful model for the study of retinal degeneration since it mimics some retinal degenerative diseases in humans, e.g., gyrate atrophy, retinitis pigmentosa or age-related macular degeneration. We evaluated the kinetics of morphological and functional changes within mice retinas injured with NaIO3 via: (1) morphological study; (2) evaluation of expression of selected neurotrophins (NTs) in injured retina; (3) visualization of proliferating and apoptotic cells; (4) electrophysiology. Our findings revealed that massive destruction of the tissue was associated with irreversible retinal dysfunction, whereas moderate retinal injury triggered regenerative mechanisms that restore bioelectrical function of the damaged retina. Next, we performed intravitreal transplantation of murine GFP+Lin- cells on the 1st day since NaIO3 administration. We analyzed number and localization of intravitreally injected GFP+Lin- cells within recipients’ retinas as well as the retinal functional changes (electroretinography). By employing stem/progenitor cell-based therapy we achieved noticeable improvement in retinal function, particularly in the case of only partial primary destruction of retinal tissue. Furthermore we investigated the neuroprotective effects of different NTs, administered intravitreally via genetically modified SCs into degenerating retinal tissue. The synthetic viral vectors based on lentivirus (LVs) backbone was used to deliver NT genes into mesenchymal stem cells isolated from bone marrow. Then, we conducted multipart analysis based on functional tests, e.g., electroretinography as well as histological, immunohistochemical, morphometric, and molecular biology studies. We found that specific, exogenously administered NTs, such as NT4/5 could effectively stimulate photoreceptor survival in the degenerating retinas. Our findings reveal that the proposed therapeutic strategy could be recommended as adjuvant therapy supporting endogenous regeneration of acute retinal damage. However, further more extensive studies are needed before the introduction of this kind of therapy into patients.
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Expression of neurotrophins and their receptors in human CD34plus bone marrow cells

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Paczkowska E., Piecyk K., Luczkowska K., Kotowski M., Roginska D., Pius-Sadowska E., Oronowicz K., Ostrowski M., Machalinski B.
Journal of Physiology and Pharmacology
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2016
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tom 67
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nr 1
3

The influence of BDNF on human umbilical cord blood stem/progenitor cells: Implications for stem cell-based therapy of neurodegenerative disorders

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Paczkowska E., Luczkowska K., Piecyk K., Roginska D., Pius-Sadowska E., Ustianowski P., Cesarska E., Dolegowska B., Celewicz Z., Machalinski B.
Acta Neurobiologiae Experimentalis
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2015
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tom 75
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nr 2
Umbilical cord blood (UCB)-derived stem/progenitor cells (SPCs) have demonstrated the potential to improve neurologic function in different experimental models. SPCs can survive after transplantation in the neural microenvironment and induce neuroprotection, endogenous neurogenesis by secreting a broad repertoire of trophic and immunomodulatory cytokines. In this study, the influence of brain-derived neurotrophic factor (BDNF) pre-treatment was comprehensively evaluated in a UCB-derived lineage-negative (Lin-) SPC population. UCB-derived Lin- cells were evaluated with respect to the expression of i) neuronal markers using immunofluorescence staining and ii) specific (TrkB) receptors for BDNF using flow cytometry. Next, after BDNF pre-treatment, Lin- cells were extensively assessed with respect to apoptosis using Western blotting and proliferation via BrdU incorporation. Furthermore, NT-3 expression levels in Lin- cells using RQ PCR and antioxidative enzyme activities were assessed. We demonstrated neuronal markers as well as TrkB expression in Lin- cells and the activation of the TrkB receptor by BDNF. BDNF pre-treatment diminished apoptosis in Lin- cells and influenced the proliferation of these cells. We observed significant changes in antioxidants as well as in the increased expression of NT-3 in Lin- cells following BDNF exposure. Complex global miRNA and mRNA profiling analyses using microarray technology and GSEA revealed the differential regulation of genes involved in the proliferation, gene expression, biosynthetic processes, translation, and protein targeting. Our results support the hypothesis that pre-treatment of stem/progenitor cells could be beneficial and may be used as an auxiliary strategy for improving the properties of SPCs.
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