Induced pluripotent stem (iPS) cells are derived from differentiated cells by different reprogramming techniques, by introducing specific transcription factors responsible for pluripotency. Induced pluripotent stem cells can serve as an excellent source for differentiated neural stem/progenitor cells (NSCs/NPs). Several methods and protocols are utilized to create a robust number of NSCs/NPs without jeopardizing the safety issues required for in vivo applications. A variety of disease-specific iPS cells have been used to study nervous system diseases. In this chapter, we will focus on some of the derivation and differentiation approaches and the application of iPS-NPs in the treatment of spinal cord injury and stroke.

• MeSH
• buněčná diferenciace * MeSH
• cévní mozková příhoda patologie   terapie   MeSH
• indukované pluripotentní kmenové buňky cytologie   MeSH
• lidé MeSH
• modely neurologické * MeSH
• nervové kmenové buňky cytologie   MeSH
• poranění míchy patologie   terapie   MeSH
• přeprogramování buněk MeSH
• transkripční faktory metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Despite the huge research into stem cells and their regenerative properties for bone healing, there are still unanswered questions including the recipient's respond to the presence of the stem cells, the fate of stem cells inside the bone defect and the possible advantage in utilizing pre-differentiated cells. To address these problems, we used human multipotent mesenchymal stromal/stem cells (MSCs), GMP Grade, in a rat model of bone formation. In a "bioreactor concept" approach seven Wistar rats were implanted with 0.2 g of synthetic bone scaffold seeded with 2 × 106 MSCs, seven Wistar rats were implanted with 0.2 g of synthetic bone scaffold seeded with 1 × 106 predifferentiated osteoblasts and 1 × 106 pre-differentiated endothelial cells and 14 Wistar rats were implanted with 0.2 g of synthetic bone scaffold without seeded cells into an intramuscular pocket on the left side of their back. The right side of each rat was used as a control, and 0.2 g of synthetic bone scaffold was implanted into the intramuscular pocket alone. To see the early stage healing the samples were harvested 14 days after the implantation, MSCs were detected by positive DAPI and MTCO2 staining in 43% of all the samples implanted with MSCs, and no inflammation signs were present in any implanted animal. New vessels could be found in both groups implanted with MSCs, but not in the control group of animals. However, hematoxylin-eosin staining could not detect newly created bone within the implant in any of the groups. These results were in line with COLL1 staining, where we could detect positive staining only in three cases, all of which were implanted with un-differentiated MSCs. According to our findings, there were no benefits of using the pre-differentiated of MSC.

• MeSH
• fyziologická neovaskularizace MeSH
• mezenchymální kmenové buňky fyziologie   MeSH
• osteoblasty fyziologie   MeSH
• osteogeneze fyziologie   MeSH
• potkani Wistar MeSH
• regenerace kostí fyziologie   MeSH
• transplantace mezenchymálních kmenových buněk metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Noninvasive cellular imaging allows the real-time tracking of grafted cells as well as the monitoring of their migration. In this review, we will focus on cell tracking using MRI, since MRI is noninvasive, clinically transferable, and displays good resolution, ranging from 50 μm in animal experiments up to 300 μm using whole body clinical scanners. In addition to information about grafted cells, MRI provides information about the surrounding tissue (i.e., lesion size, edema, inflammation), which may negatively affect graft survival or the functional recovery of the tissue. Transplanted cells are labeled with MR contrast agents in vitro prior to transplantation in order to visualize them in the host tissue. The chapter will focus on the use of superparamagnetic iron oxide nanoparticles (SPIO), because they have strong effects on T2 relaxation yet do not affect cell viability, and will provide an overview of different modifications of SPIO and their use in MR tracking in living organisms.

• MeSH
• barvení a značení metody   MeSH
• buněčná diferenciace MeSH
• buněčný tracking metody   MeSH
• dextrany chemie   MeSH
• ferrokyanidy analýza   MeSH
• kontrastní látky chemie   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• magnetická rezonanční tomografie metody   MeSH
• magnetické nanočástice chemie   MeSH
• magnetismus metody   MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• myši MeSH
• pohyb buněk MeSH
• transplantace mezenchymálních kmenových buněk MeSH
• viabilita buněk MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Noninvasive cellular imaging allows the real-time tracking of grafted cells as well as the monitoring of their migration. Several techniques for in vivo cellular imaging are available that permit the characterization of transplanted cells in a living organism, including magnetic resonance imaging (MRI), bioluminescence, positron emission tomography, and multiple photon microscopy. All of these methods, based on different principles, provide distinctive, usually complementary information. In this review, we will focus on cell tracking using MRI, since MRI is noninvasive, clinically transferable, and displays good resolution, ranging from 50microm in animal experiments up to 300microm using whole body clinical scanners. In addition to information about grafted cells, MRI provides information about the surrounding tissue (i.e., lesion size, edema, inflammation), which may negatively affect graft survival or the functional recovery of the tissue. Transplanted cells are labeled with MR contrast agents in vitro prior to transplantation in order to visualize them in the host tissue. The chapter will focus on the use of superparamagnetic iron oxide nanoparticles (SPIO), because they have strong effects on T2 relaxation yet do not affect cell viability, and will provide an overview of different modifications of SPIO and their use in MR tracking in living organisms.

• MeSH
• buněčné kultury MeSH
• financování organizované MeSH
• kontrastní látky chemie   metabolismus   MeSH
• kovové nanočástice chemie   MeSH
• krysa rodu rattus MeSH
• kultivované buňky MeSH
• lidé MeSH
• magnetická rezonanční tomografie metody   přístrojové vybavení   MeSH
• mezenchymální kmenové buňky cytologie   fyziologie   MeSH
• mícha anatomie a histologie   metabolismus   patologie   MeSH
• mozek anatomie a histologie   metabolismus   patologie   MeSH
• neurodegenerativní nemoci metabolismus   patologie   MeSH
• transplantace mezenchymálních kmenových buněk MeSH
• železité sloučeniny chemie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH

Our study followed the changes in thalamic nuclei metabolism, hindlimb sensitivity to thermal stimulation, and locomotor function after spinal cord injury (SCI). MR spectroscopy (MRS) was used to examine the thalamic nuclei of rats 1 day before and 1, 3, 6, and 15 days after SCI or sham surgery. All animals were tested before MRS measurements for motor performance and thermal sensitivity. SCI induced by balloon compression caused complete paraplegia from the first to third day, followed by partial functional recovery during the second week. MRS revealed an increase in N-acetylaspartate (NAA) concentration in the thalamic nuclei on the first day after SCI, which decreased by the third day. The data also showed an increase in inositol (Ins), glutamate, and creatine (Cr) concentrations on the third day postinjury; the Ins concentration remained elevated on the sixth day. In sham-operated animals an increase in NAA concentration was observed on the sixth and fifteenth days after surgery and an increase in Cr concentration on the third day. A positive correlation between Ins concentration and hindlimb sensitivity in both SCI and sham-operated animals suggests changes in glial activity, while changes in NAA levels may indicate the response of thalamic neuronal cells to injury. (c) 2008 Wiley-Liss, Inc.

• MeSH
• analýza rozptylu MeSH
• cholin metabolismus   MeSH
• financování organizované MeSH
• inositol metabolismus   MeSH
• kreatin metabolismus   MeSH
• krysa rodu rattus MeSH
• kyselina aspartová analogy a deriváty   metabolismus   MeSH
• kyselina glutamová metabolismus   MeSH
• magnetická rezonanční spektroskopie metody   MeSH
• poranění míchy metabolismus   MeSH
• potkani Wistar MeSH
• protony MeSH
• thalamus metabolismus   MeSH
• vysoká teplota MeSH
• zadní končetina fyziologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH

Neurogenic pulmonary edema is an acute life-threatening complication following central nervous system injury. The exact pathogenic mechanism leading to its development is still unclear. We introduce a new hypothesis that high levels of anesthesia might protect the organism against the development of neurogenic pulmonary edema due to a more pronounced inhibition of the hypothalamic, brainstem and spinal vasoactive sympathetic centers. On the basis of a more pronounced neuronal inhibition of the vasoactive centers, a severe sympathetic discharge does not occur and neurogenic pulmonary edema does not develop. In contrast, an insufficient anesthesia level is not able to inhibit the sympathetic nervous system during an injury of the central nervous system and thus neurogenic pulmonary edema develops. During experiments with central nervous system injury, low-anesthesia-induced neurogenic pulmonary edema might negatively influence the overall recovery of the animal. More importantly, during a neurosurgical intervention, insufficient anesthesia might similarly lead to neurogenic pulmonary edema development in operated patients. Our hypothesis indicates the necessity of precisely monitoring of the level anesthesia during experimental manipulations of the central nervous system in animals or neurosurgical interventions in humans.

• MeSH
• anestezie metody   škodlivé účinky   MeSH
• biologické modely MeSH
• centrální nervový systém patofyziologie   zranění   MeSH
• financování organizované MeSH
• intrakraniální hypertenze patofyziologie   MeSH
• lidé MeSH
• plicní edém etiologie   patofyziologie   MeSH
• sympatický nervový systém patofyziologie   MeSH
• vazomotorický systém patofyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH

We describe a new model of neurogenic pulmonary edema in spinal cord injured Wistar male rats. The pulmonary edema was elicited by an epidural thoracic balloon compression spinal cord lesion, performed under a low concentration of isoflurane (1.5 or 2%) in air. Anesthesia with 1.5% isoflurane promoted very severe interstitial and intraalveolar neurogenic pulmonary edema with a significantly increased thickness of the alveolar walls and massive pulmonary hemorrhage. In this group, 33% of animals died. Anesthesia with 2% isoflurane promoted severe interstitial and intraalveolar neurogenic pulmonary edema with less thickening of the alveolar walls and pulmonary hemorrhage. For evoking severe neurogenic pulmonary edema in spinal cord injured rats, 2% isoflurane anesthesia would be more suitable. However, if very severe neurogenic pulmonary edema needs to be evoked, spinal cord injury under 1.5% isoflurane anesthesia could be used, but one-third of the animals will be lost.

• MeSH
• anestetika inhalační aplikace a dávkování   MeSH
• financování organizované MeSH
• isofluran aplikace a dávkování   MeSH
• krysa rodu rattus MeSH
• modely nemocí na zvířatech MeSH
• plicní edém etiologie   patologie   MeSH
• poranění míchy komplikace   MeSH
• potkani Wistar MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH

Anesthetics can either promote or inhibit the development of neurogenic pulmonary edema (NPE) after central nervous system (CNS) injury. The influence of isoflurane was examined in male Wistar rats using 1.5%, 2%, 2.5%, 3%, 4%, or 5% isoflurane in air. Epidural balloon compression of the thoracic spinal cord was performed. The development of NPE was examined in vivo and on histologic sections of lung tissue. Animals anesthetized with 1.5% or 3% isoflurane were behaviorally monitored using the BBB and plantar tests for 7 weeks post-injury. The spinal cord was examined using MRI and morphometry of the spared white and gray matter. All animals from the 1.5% and 2% groups developed NPE. Almost 42% of the animals in the 1.5% group died of severe pulmonary hemorrhage and suffocation; x-rays, the pulmonary index, and the histological picture revealed a massive NPE. More than 71% of the animals from the 2.5% and 3% groups did not develop any signs of NPE. Blood pressure after spinal cord compression rose more in the 1.5% group than in the 3% one. In the 1.5% group, the sympathetic ganglionic blockade prevented the neurogenic pulmonary edema development. Animals from the 3% group recovered behaviorally more rapidly than did the animals from the 1.5% group; morphometry and MRI of the lesions showed no differences. Thus, low levels of isoflurane anesthesia promote NPE in rats with a compressed spinal cord and significantly complicates their recovery. The optimal concentration of anesthesia for performing a spinal cord compression lesion is 2.5-3% isoflurane in air.

• MeSH
• anestetika inhalační aplikace a dávkování   škodlivé účinky   MeSH
• financování organizované MeSH
• isofluran aplikace a dávkování   škodlivé účinky   MeSH
• krevní tlak účinky léků   MeSH
• krysa rodu rattus MeSH
• plicní edém chemicky indukované   patologie   MeSH
• poranění míchy komplikace   MeSH
• potkani Wistar MeSH
• srdeční frekvence účinky léků   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH

1. Emerging clinical studies of treating brain and spinal cord injury (SCI) led us to examine the effect of autologous adult stem cell transplantation as well as the use of polymer scaffolds in spinal cord regeneration. We compared an intravenous injection of mesenchymal stem cells (MSCs) or the injection of a freshly prepared mononuclear fraction of bone marrow cells (BMCs) on the treatment of an acute or chronic balloon-induced spinal cord compression lesion in rats. Based on our experimental studies, autologous BMC implantation has been used in a Phase I/II clinical trial in patients (n=20) with a transversal spinal cord lesion. 2. MSCs were isolated from rat bone marrow by their adherence to plastic, labeled with iron-oxide nanoparticles and expanded in vitro. Macroporous hydrogels based on derivatives of 2-hydroxyethyl methacrylate (HEMA) or 2-hydroxypropyl methacrylamide (HPMA) were prepared, then modified by their copolymerization with a hydrolytically degradable crosslinker, N,O-dimethacryloylhydroxylamine, or by different surface electric charges. Hydrogels or hydrogels seeded with MSCs were implanted into rats with hemisected spinal cords. 3. Lesioned animals grafted with MSCs or BMCs had smaller lesions 35 days postgrafting and higher scores in BBB testing than did control animals and also showed a faster recovery of sensitivity in their hind limbs using the plantar test. The functional improvement was more pronounced in MSC-treated rats. In MR images, the lesion populated by grafted cells appeared as a dark hypointense area and was considerably smaller than in control animals. Morphometric measurements showed an increase in the volume of spared white matter in cell-treated animals. In the clinical trial, we compared intraarterial (via a. vertebralis, n=6) versus intravenous administration of BMCs (n=14) in a group of subacute (10-33 days post-SCI, n=8) and chronic patients (2-18 months, n=12). For patient follow-up we used MEP, SEP, MRI, and the ASIA score. Our clinical study revealed that the implantation of BMCs into patients is safe, as there were no complications following cell administration. Partial improvement in the ASIA score and partial recovery of MEP or SEP have been observed in all subacute patients who received cells via a. vertebralis (n=4) and in one out of four subacute patients who received cells intravenously. Improvement was also found in one chronic patient who received cells via a. vertebralis. A much larger population of patients is needed before any conclusions can be drawn. The implantation of hydrogels into hemisected rat spinal cords showed that cellular ingrowth was most pronounced in copolymers of HEMA with a positive surface electric charge. Although most of the cells had the morphological properties of connective tissue elements, we found NF-160-positive axons invading all the implanted hydrogels from both the proximal and distal stumps. The biodegradable hydrogels degraded from the border that was in direct contact with the spinal cord tissue. They were resorbed by macrophages and replaced by newly formed tissue containing connective tissue elements, blood vessels, GFAP-positive astrocytic processes, and NF-160-positive neurofilaments. Additionally, we implanted hydrogels seeded with nanoparticle-labeled MSCs into hemisected rat spinal cords. Hydrogels seeded with MSCs were visible on MR images as hypointense areas, and subsequent Prussian blue histological staining confirmed positively stained cells within the hydrogels. 4. We conclude that treatment with different bone marrow cell populations had a positive effect on behavioral outcome and histopathological assessment after SCI in rats; this positive effect was most pronounced following MSC treatment. Our clinical study suggests a possible positive effect in patients with SCI. Bridging the lesion cavity can be an approach for further improving regeneration. Our preclinical studies showed that macroporous polymer hydrogels based on derivatives of HEMA or HPMA are suitable materials for bridging cavities after SCI; their chemical and physical properties can be modified to a specific use, and 3D implants seeded with different cell types may facilitate the ingrowth of axons.

• MeSH
• autologní transplantace MeSH
• buňky kostní dřeně fyziologie   MeSH
• hydrogely * terapeutické užití   MeSH
• komprese míchy terapie   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• mezenchymální kmenové buňky fyziologie   MeSH
• monocyty transplantace   MeSH
• pohyb buněk MeSH
• polymery terapeutické užití   MeSH
• poranění míchy * terapie   MeSH
• regenerace nervu MeSH
• transplantace kostní dřeně * metody   MeSH
• transplantace mezenchymálních kmenových buněk * metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

Stem cell transplants into spinal cord lesions may help to improve regeneration and spinal cord function. Clinical studies are necessary for transferring preclinical findings from animal experiments to humans. We investigated the transplantation of unmanipulated autologous bone marrow in patients with transversal spinal cord injury (SCI) with respect to safety, therapeutic time window, implantation strategy, method of administration, and functional improvement. We report data from 20 patients with complete SCI who received transplants 10 to 467 days postinjury. The follow-up examinations were done at 3, 6, and 12 months after implantation by two independent neurologists using standard neurological classification of SCI, including the ASIA protocol, the Frankel score, the recording of motor and somatosensory evoked potentials, and MRI evaluation of lesion size. We compared intra-arterial (via catheterization of a. vertebralis) versus intravenous administration of all mononuclear cells in groups of acute (10-30 days post-SCI, n=7) and chronic patients (2-17 months postinjury, n=13). Improvement in motor and/or sensory functions was observed within 3 months in 5 of 6 patients with intra-arterial application, in 5 of 7 acute, and in 1 of 13 chronic patients. Our case study shows that the implantation of autologous bone marrow cells appears to be safe, as there have been no complications following implantation to date (11 patients followed up for more than 2 years), but longer follow-ups are required to determine that implantation is definitively safe. Also, we cannot yet confirm that the observed beneficial effects were due to the cell therapy. However, the outcomes following transplantation in acute patients, and in one chronic patient who was in stable condition for several months prior to cell implantation, are promising. It is evident that transplantation within a therapeutic window of 3-4 weeks following injury will play an important role in any type of stem cell SCI treatment. Trials involving a larger population of patients and different cell types are needed before further conclusions can be drawn.

• MeSH
• akutní nemoc MeSH
• autologní transplantace MeSH
• chronická nemoc MeSH
• dospělí MeSH
• elektrofyziologie MeSH
• lidé MeSH
• magnetická rezonanční tomografie MeSH
• následné studie MeSH
• obnova funkce fyziologie   MeSH
• poranění míchy chirurgie   patofyziologie   patologie   MeSH
• regenerace nervu fyziologie   MeSH
• transplantace kostní dřeně * metody   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH