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
The biological effects of several superparamagnetic iron oxide nanoparticles (SPIONs) varying in their surface coating were tested using human bone marrow mesenchymal stromal cells from two donors - hBMSCs-1 and hBMSCs-2. The measurements were performed at two intervals - after 72 h exposure to the nanoparticles and after an additional 72 h cell growth without nanoparticles. The dose of SPIONs used (15.4 μg Fe/ml) was selected as being sufficient for in vivo cell tracking using magnetic resonance imaging (MRI). Concerning cell viability and cell death, only the hBMSCs-2 seemed to be sensitive to the action of SPIONs. However, an increase of oxidative injury to lipids, proteins and DNA as a consequence of exposure to SPIONs was detected in cells from both donors. Particularly the levels of lipid peroxidation were high and increased further with time, regardless of the type of nanoparticle. Lowering intracellular label concentrations and authenticating oxidative stress levels using in vivo experiments are required to ensure the safety of SPIONs for biomedical applications.
- MeSH
- buněčná smrt účinky léků MeSH
- dítě MeSH
- lidé středního věku MeSH
- lidé MeSH
- magnetická rezonanční tomografie MeSH
- magnetické nanočástice škodlivé účinky MeSH
- mezenchymální kmenové buňky účinky léků MeSH
- oxidační stres účinky léků MeSH
- peroxidace lipidů účinky léků MeSH
- poškození DNA účinky léků MeSH
- proteiny účinky léků MeSH
- viabilita buněk účinky léků MeSH
- železité sloučeniny škodlivé účinky MeSH
- Check Tag
- dítě MeSH
- lidé středního věku MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Multimodal imaging-therapeutic nanoprobe TiO(2)@RhdGd was prepared and successfully used for in vitro and in vivo cell tracking as well as for killing of cancer cells in vitro. TiO(2) nanoparticles were used as a core for phosphonic acid modified functionalities, responsible for contrast in MRI and optical imaging. The probe shows high (1)H relaxivity and relaxivity density values. Presence of fluorescent dye allows for visualization by means of fluorescence microscopy. The applicability of the probe was studied, using mesenchymal stem cells, cancer HeLa cells, and T-lymphocytes. The probe did not exhibit toxicity in any of these systems. Labeled cells were successfully visualized in vitro by means of fluorescence microscopy and MRI. Furthermore, it was shown that the probe TiO(2)@RhdGd can be changed into a cancer cell killer upon UV light irradiation. The above stated results represent a valuable proof of a principle showing applicability of the probe design for diagnosis and therapy.
- MeSH
- afinitní značky chemická syntéza chemie farmakologie MeSH
- antitumorózní látky chemická syntéza chemie farmakologie MeSH
- fluorescenční barviva chemická syntéza chemie farmakologie MeSH
- fluorescenční mikroskopie MeSH
- gadolinium MeSH
- HeLa buňky MeSH
- komplexní sloučeniny chemická syntéza chemie farmakologie MeSH
- kyseliny fosforité chemická syntéza chemie farmakologie MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- magnetická rezonanční tomografie MeSH
- mezenchymální kmenové buňky metabolismus MeSH
- myši MeSH
- nanočástice MeSH
- T-lymfocyty metabolismus MeSH
- titan chemie farmakologie MeSH
- ultrafialové záření MeSH
- viabilita buněk účinky léků MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
During the last decade, much progress has been made in developing protocols for the differentiation of human embryonic stem cells (hESCs) into a neural phenotype. The appropriate agent for cell therapy is neural precursors (NPs). Here, we demonstrate the derivation of highly enriched and expandable populations of proliferating NPs from the CCTL14 line of hESCs. These NPs could differentiate in vitro into functionally active neurons, as confirmed by immunohistochemical staining and electrophysiological analysis. Neural cells differentiated in vitro from hESCs exhibit broad cellular heterogeneity with respect to developmental stage and lineage specification. To analyze the population of the derived NPs, we used fluorescence-activated cell sorting (FACS) and characterized the expression of several pluripotent and neural markers, such as Nanog, SSEA-4, SSEA-1, TRA-1-60, CD24, CD133, CD56 (NCAM), beta-III-tubulin, NF70, nestin, CD271 (NGFR), CD29, CD73, and CD105 during long-term propagation. The analyzed cells were used for transplantation into the injured rodent brain; the tumorigenicity of the transplanted cells was apparently eliminated following long-term culture. These results complete the characterization of the CCTL14 line of hESCs and provide a framework for developing cell selection strategies for neural cell-based therapies.
- MeSH
- biologické markery metabolismus MeSH
- buněčná diferenciace MeSH
- buněčné linie MeSH
- buněčný rodokmen MeSH
- embryonální kmenové buňky cytologie MeSH
- fenotyp MeSH
- imunohistochemie MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- mozek metabolismus patologie MeSH
- neurony cytologie metabolismus transplantace MeSH
- průtoková cytometrie MeSH
- tkáňová terapie - dějiny MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
