- Klíčová slova
- lipomyelomeningokéla, brnění,
- MeSH
- bolest etiologie MeSH
- cauda equina patologie růst a vývoj MeSH
- defekty neurální trubice chirurgie diagnostické zobrazování etiologie klasifikace komplikace patologie MeSH
- dítě MeSH
- dolní končetina patologie MeSH
- laminektomie klasifikace metody MeSH
- lidé MeSH
- lipom MeSH
- magnetická rezonanční tomografie MeSH
- meningomyelokéla * chirurgie patologie MeSH
- mícha patologie růst a vývoj MeSH
- páteř patologie růst a vývoj MeSH
- páteřní kanál chirurgie patologie MeSH
- předoperační vyšetření MeSH
- výsledek terapie MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Expression of doublecortin (DCX), a 43-53 kDa microtubule binding protein, is frequently used as (i) an early neuronal marker to identify the stage of neuronal maturation of in vivo grafted neuronal precursors (NSCs), and (ii) a neuronal fate marker transiently expressed by immature neurons during development. Reliable identification of the origin of DCX-immunoreactive cells (i.e., host vs. graft) requires detailed spatial and temporal mapping of endogenous DCX expression at graft-targeted brain or spinal cord regions. Accordingly, in the present study, we analyzed (i) the time course of DCX expression in pre- and postnatal rat and porcine spinal cord, and (ii) the DCX expression in spinally grafted porcine-induced pluripotent stem cells (iPS)-derived NSCs and human embryonic stem cell (ES)-derived NSCs. In addition, complementary temporospatial GFAP expression study in porcine spinal cord was also performed. In 21-day-old rat fetuses, an intense DCX immunoreactivity distributed between the dorsal horn (DH) and ventral horn was seen and was still present in the DH neurons on postnatal day 20. In animals older than 8 weeks, no DCX immunoreactivity was seen at any spinal cord laminae. In contrast to rat, in porcine spinal cord (gestational period 113-114 days), DCX was only expressed during the pre-natal period (up to 100 days) but was no longer present in newborn piglets or in adult animals. Immunohistochemical analysis was confirmed with a comparable expression profile by western blot analysis. Contrary, the expression of porcine GFAP started within 70-80 days of the pre-natal period. Spinally grafted porcine iPS-NSCs and human ES-NSCs showed clear DCX expression at 3-4 weeks postgrafting. These data indicate that in spinal grafting studies which employ postnatal or adult porcine models, the expression of DCX can be used as a reliable marker of grafted neurons. In contrast, if grafted neurons are to be analyzed during the first 4 postnatal weeks in the rat spinal cord, additional markers or grafted cell-specific labeling techniques need to be employed to reliably identify grafted early postmitotic neurons and to differentiate the DCX expression from the neurons of the host.
- MeSH
- časové faktory MeSH
- druhová specificita MeSH
- embryonální kmenové buňky metabolismus transplantace MeSH
- indukované pluripotentní kmenové buňky metabolismus transplantace MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- lidé MeSH
- mícha růst a vývoj metabolismus MeSH
- neurogeneze fyziologie MeSH
- neuropeptidy biosyntéza MeSH
- potkani Wistar MeSH
- prasata MeSH
- proteiny asociované s mikrotubuly biosyntéza MeSH
- těhotenství MeSH
- transplantace kmenových buněk trendy MeSH
- vývojová regulace genové exprese * MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The metabolic form of vitamin A, retinol, has a pivotal role in the nervous system development and neuronal differentiation, both during embryogenesis through maternal-fetal support and in the early postnatal life. Retinoic acid was administered orally at a dose of 10 mg/kg body weight to pregnant female rats through days 8-10 of gestation. Spinal cord sections were processed for histochemical visualization one day after birth and on day 21, when weaning is expected. NADPH-diaphorase (NADPH-d)-positive neurons were found in the dorsal horn, around the central canal, and at the intermediolateral cell column on postnatal days 1 and 21 in both control and experimental groups. There were no NADPHd-positive structures in the ventral horn. The results suggest that prenatal administration of high doses of retinoic acid is not associated with postnatal morphological changes in NADPH-d-positive neurons in the rat spinal cord. Levels of antioxidants and related enzymes in retinoid storage organs were measured to estimate possible side effects. The activities of enzymes detoxifying superoxide radicals and peroxides were supressed after birth. A decrease in the level of reduced glutathione was observed on postnatal day 21, indicating an unbalanced redox environment.
- MeSH
- játra účinky léků růst a vývoj metabolismus MeSH
- krysa rodu rattus MeSH
- mícha účinky léků růst a vývoj metabolismus MeSH
- NADP metabolismus MeSH
- potkani Wistar MeSH
- těhotenství MeSH
- vitamin A aplikace a dávkování farmakologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Currently, there is no effective strategy for the treatment of spinal cord injury (SCI). A suitable combination of modern hydrogel materials, modified to effectively bridge the lesion cavity, combined with appropriate stem cell therapy seems to be a promising approach to repair spinal cord damage. We demonstrate the synergic effect of porosity and surface modification of hydrogels on mesenchymal stem cell (MSC) adhesiveness in vitro and their in vivo survival in an experimental model of SCI. MSCs were seeded on four different hydrogels: hydroxypropylmethacrylate-RGD prepared by heterophase separation (HPMA-HS-RGD) and three other hydrogels polymerized in the presence of a solid porogen: HPMA-SP, HPMA-SP-RGD, and hydroxy ethyl methacrylate [2-(methacryloyloxy)ethyl] trimethylammonium chloride (HEMA-MOETACl). Their adhesion capability and cell survival were evaluated at 1, 7, and 14 days after the seeding of MSCs on the hydrogel scaffolds. The cell-polymer scaffolds were then implanted into hemisected rat spinal cord, and MSC survival in vivo and the ingrowth of endogenous tissue elements were evaluated 1 month after implantation. In vitro data demonstrated that HEMA-MOETACl and HPMA-SP-RGD hydrogels were superior in the number of cells attached. In vivo, the highest cell survival was found in the HEMA-MOETACl hydrogels; however, only a small ingrowth of blood vessels and axons was observed. Both HPMA-SP and HPMA-SP-RGD hydrogels showed better survival of MSCs compared with the HPMA-HS-RGD hydrogel. The RGD sequence attached to both types of HPMA hydrogels significantly influenced the number of blood vessels inside the implanted hydrogels. Further, the porous structure of HPMA-SP hydrogels promoted a statistically significant greater ingrowth of axons and less connective tissue elements into the implant. Our results demonstrate that the physical and chemical properties of the HPMA-SP-RGD hydrogel show the best combination for bridging a spinal cord lesion, while the HEMA-MOETACl hydrogel serves as the best carrier of MSCs.
- MeSH
- axony účinky léků fyziologie MeSH
- buněčná adheze MeSH
- cholin analogy a deriváty chemie farmakologie MeSH
- fyziologická neovaskularizace MeSH
- hydrogely chemie farmakologie MeSH
- kmenové buňky cytologie účinky léků fyziologie MeSH
- krysa rodu rattus MeSH
- methakryláty chemie farmakologie MeSH
- mícha krevní zásobení účinky léků růst a vývoj patologie MeSH
- oligopeptidy chemie farmakologie MeSH
- poranění míchy terapie MeSH
- poréznost MeSH
- potkani Wistar MeSH
- regenerace nervu účinky léků MeSH
- tkáňové podpůrné struktury MeSH
- transplantace kmenových buněk MeSH
- viabilita buněk MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- MeSH
- bederní obratle MeSH
- financování organizované MeSH
- histocytologické preparační techniky MeSH
- hrudní obratle MeSH
- lumbosakrální krajina MeSH
- mícha anatomie a histologie cytologie růst a vývoj MeSH
- mikroskopie MeSH
- NADPH-dehydrogenasa MeSH
- neurony MeSH
- neurotransmiterové látky MeSH
- nitrátreduktasa (NADPH) MeSH
- oxid dusnatý metabolismus MeSH
- potkani Wistar MeSH
- synthasa oxidu dusnatého MeSH
- věkové faktory MeSH
- zmrazené řezy MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
