Objective: Demyelination of CNS axons occurs in a number of pathological conditions, including multiple sclerosis and spinal cord injury. The demyelination can be repaired by cell therapy in both humans and rodents, even within the CNS, remyelination can be achieved by neural stem cells (NSCS) implantation.Materials and Methods: In this study, in order to test whether the transplantation of NSCS affect remyelination efficiency in rat demyelination model or not, first focal demyelinating lesions were done in the adult rat spinal cord with ethidium bromide then we prepared NSCS derived bone marrow stromal cells and transplanted them to rat demyelination model.Results: These results indicate that NSCS are capable of differentiation to myelinating cells which eliciting remyelination and have invaluable applications in treatment of neurodegenerative diseases such as spinal cord injury.Conclusion: BMSCs were efficiently induced into neurospheres and the NSCS were prepared and the result of transplantation indicated that NSCS are capable of differentiation to myelinating cells which eliciting remyelination.

BACKGROUND AND OBJECTIVE: The use of an appropriate stimulus to speed up in vitro and in vivo stem cell proliferation and growth is the one of the most important issues in cell therapy. The aim of this study was in vitro evaluation of hydroethanolic extraction of citrus aurantium on proliferation and growth rates of neonate rat neural stem cells.METHODS: Neural stem cells were isolated using enzymatic digestion from hippocampus region of 3 day old neonatal rat brain. To determine optimal concentration of citrus aurantium extraction, isolated neural stem cells were treated at 200, 400, 600, 800 and 1000 mg/ml for 48 h. Then cells proliferation rate were evaluated using MTT assay method.FINDINGS: Citrus aurantium promoted neural stem cells proliferation compared to untreated cells. As well as the effect of aurantium extract is dose dependent. The result of this study showed that proliferative rate in treated cells with 600 mg/ml of Citrus aurantium extraction (0.81±0.06) significantly is higher than control (untreated neural stem cells) group (0.59±0.02). (p<0.05)CONCLUSION: Regarding the effect of Citrus aurantium extract on the proliferation rate of neural stem cells, the use of hydroethanolic extraction of Citrus aurantium can be useful for clinical purposes to cell replacement therapy of various neurodegenerative disorders such as spinal cord injury.

Objective (s): The stimulation of neural stem cells (NSCS) differentiation into neurons has attracted great attention in management of neurodegenerative disease and traumatic brain injury. It has been reported that selegiline could enhance the morphologic differentiation of embryonic stem cells.Therefore this study aimed to investigate the effects of selegiline on NSCS differentiation with focus on the role of neurotrophic factor gene expression.Materials and Methods: The NSCS were isolated from lateral ventricle of C57 mice brain. The cells were exposed to selegiline in nano to micromolar concentrations for 24 hr or 72 hr. In order to assay the effect of selegiline on NSCS differentiation into neurons, astrocytes and oligodendrocytes, immunocytochemical techniques were utilized. Samples were exposed to specific antibodies against neurons (b tubulin), astrocytes (GFAP) and oligodendrocytes (OSP). The expression of BDNF, NGF and NT3 genes was investigated using Real‐Time PCR.Results: Our findings revealed that selegiline increased NSCS differentiation into neurons at 10‐7 and 10‐8 M and decreased the differentiation into astrocytes at 10‐9, while oligodendrocyte did not significantly change in any of the used concentrations. In addition data analyses showed that selegiline increased BDNF, NGF and NT3 gene expression at 24 hr, but did not change them in the other time of exposure (72 hr) except 10‐7 M concentration of selegiline, which increased NT3 expression.Conclusion: Our results indicate selegiline induced the differentiation of NSCS into neurons and in this context the role of neurotrophic factors is important and should be considered.

Objective: Assessment of functional recovery of neural stem cells (NSCS) and (-)-deprenyl, in a contusive spinal cord injury model in rats.Materials and Methods: A total of 24 female Sprague Dawley rats were randomly, but equally (n=6), allocated into the following groups: control, sham, NSC graft and NSC graft + (-)-deprenyl. All animals were laminectomized at the T13 level. Contusion was performed according to the weight dropping technique in the control, NSC and NSC graft + (-)-deprenyl groups. Daily injections of 0.1 mg/kg (ip) (-)-deprenyl were administered to the NSC graft + (-)-deprenyl group and an equal amount of saline into the other groups for 14 days. The NSC graft and NSC graft + (-)-deprenyl groups received stereotaxic injections of 100,000 labelled NSCS at day nine after injury. Behavioral Basso, Beattie and Bresnahan (BBB) test was carried out in all groups at day one (after the contusion day) and at the end of each week for eight weeks. In addition, cavity and spared tissue volume at the site of injury and number of motoneurons at frozen sections of spinal cord were obtained and compared by ANOVA. Differentiation of grafted NSCS into astrocytes, oligodendrocytes and neurons were evaluated by immunohistochemistry. Results: Motor ability of the NSC graft + (-)-deprenyl group in comparison with the control and NSCS groups increased significantly at the end of the study. The mean volume of spared spinal cord and mean number of motoneurons significantly increased in the NSCS and NSC graft + (-)-deprenyl groups compared with the control group. Immunohistochemical evaluations revealed that the grafted NSCS were alive at the end of the study and differentiated into astrocyes, oligodendrocytes and neurons in both the NSCS and NSC graft + (-)-deprenyl groups. In addition, in the NSCS graft group, transplanted cells mainly concentrated around the cavity and showed less differentiation, while in the NSCS graft + (-)-deprenyl group transplanted cells were more scattered and differentiated into one of the above mentioned cell lines. Conclusion: The results of the present study indicate that (-)-deprenyl and NSCS, probably via protection of motoneurons, spinal cord tissue and replacement of lost cells, improves motor recovery in a contusive SCI model in rats.

Introduction Earthquake is one of the most important risks in high seismic countries like Iran and, considering solutions to reduce the seismic vulnerability of buildings is one of priorities for designers, builders, and regulations in this area. Experiences of past earthquakes show that a significant part of the human injuries and financial loses is due to damage to NSCS in buildings - including architectural, mechanical and electrical components. Tall buildings are more important in reducing NSC’s seismic vulnerability due to their size, large number of residences and occupancies and the special characteristics of their structures. If earthquake happen,  even if it does not cause serious damage to the structural stability of tall buildings, any damage to their NSCS such as the facade, stairs, facilities, interior decoration, etc., in addition to the possibility of causing physical injuries and loss of life, It can leave significant costs for reconstruction. Managing these damages includes awareness, prevention and reduction of their vulnerability before the earthquake occurred and in the process of designing and constructing buildings. Research Necessity Tall buildings, due to their height and scale, have a considerable impact on the urban landscape, and for this reason, designers mostly like to use complex forms with complicated and impressive geometric in the design of these buildings. However, architectural forms play a major role in the seismic performance of tall buildings. This research explores the effect of the “Twisted forms” as a complex architectural form, with various plans and different heights on the seismic performance of tall buildings with steel diagrid structural system. The seismic performance indicators are drift and lateral displacement as indicators of measuring the vulnerability of NSCS and the weight per unit area of the structure as an indicator of the optimal structural design of buildings Research Method  This research is of a quantitative type and has been done by computer simulation and analysis method. For this purpose, first, the initial models with triangle, square, hexagon, and circle plans and with various heights including 160, 180, and 200 meters were made parametrically in Rhino software and the Grasshopper plugin. Then, twisted forms are made by twisting the floors of the primary models relative to each other by 1, 2, and 3 degrees. After that, the diagrid structural system has been designed for all of the architectural forms directly in the parametric design environment using the Karamba3D finite element analysis plugin. The linear static analysis for serviceability earthquake load has been used for designing and tuning the diagrid structures Finally, the seismic performance indicators including drift and lateral displacement as the indicators of measuring the vulnerability of NSCS and the weight per unit area of the structure as the indicator of the optimal structural design of tall buildings are extracted and analyzed using charts and tables. Innovations and Results The effects of simple various architectural forms on the seismic performance of tall buildings, have been studied in previous researches including taper, concave and convex forms. However, so far, complex forms have been less investigated. This research explores the effect of the “Twisting forms” as a complex architectural form, with various plans and different heights on the seismic performance of tall buildings with steel diagrid structural system. This issue has not been investigated in previous studies. The results show that in the diagrid structure with steel materials at the heights and plans investigated in this research, the twisting of the floors in the height increases the drift, lateral displacement, and weight per unit area of ​​the structure in the forms with triangular and quadrilateral plans in all 160,180 and 200 meters height. As a result, choosing these types of plans for twisting forms can reduce the lateral stability of tall buildings and increase the probability of damage to NSCS during an earthquake. On the other hand, by increasing the number of sides of the plan (forms with circular and hexagonal plans), the relative twist of the floors in terms of degrees (from zero to 3 degrees) has not cause significant changes in any of evaluated indicators. Resultantly, using plans with more sides for tall buildings with twisted forms would make it possible to avoid the adverse effects of twisting and to design optimal tall buildings from a seismic point of view.

Objective: To evaluate the potential of Neural Stem Cell (NSC)-based approach to correct the metabolic defect and to ameliorate pathology in Twitcher (Twi) mice, a true model of Globoid Cell Leukodystrophy (GLD).Materials and Methods: Twi mice display severe demyelination and neurodegeneration, with a median survival of 46 days. We derived NSC lines from the subventricular zone of neonatal Twi mice and of wt littermates. Using bidirectional lentiviral vectors (bdLV) encoding for galactocerebrosidase (GALC) and GFP, we achieved high efficiency of transduction (>80%) and supraphysiological GALC levels (2-3 fold the wt levels) in mutant NSC, with no toxicity or functional impairment due to transgene over-expression. Similar supraphysiological GALC levels were obtained following bdLV. GALC transduction of wt NSC. Of importance, gene-corrected cells allowed more efficient metabolic cross-correction of GALC-deficient NSC then wt cells in vitro, due to more efficient enzyme secretion in the extracellular milieau. In order to obtain a stable source of GALC-secreting cells in the brain we transplanted wt or GALC over-expressing NSCS into the telencephalic lateral ventricles of neonatal (post-natal day 2) Twi mice (1x10^6 total cells, bilateral injection). Forty days after transplant we evaluated on NSC-treated mice and on untreated controls: i) NSC engraftment, distribution and fate (by immunocytochemistry); ii) the presence of a functional GALC protein in tissues and cerebrospinal fluid (by western blot and enzymatic assays); iii) intracellular storage (by lectin staining); iv) improvement of pathology (by immunocytochemistry and qPCR ). A group of treated mice was monitored for body weight and motor skills until a fixed human endpoint, in order to assess the impact of NSC therapy in delaying the onset of symptoms and prolonging lifespan.Results: Forty days after transplant we found NSC (identified by GFP expression) widely distributedinto the brain parenchyma, from the olfactory bulb to the hippocampus. Many cells were found lining the ventricles, along the corpus callosum and in the external capsula. Engrafted NSC (1-3% of the total injected cells) either expressed glial cell markers or retained antigenic features of immature neural cells and did not show proliferative activity. Of note, they robustly produced and secreted the GALC protein, as demonstrated by immunohistochemistry and by western blot using anti-GALC antibody. Most important, GALC activity was restored to 50% of wt levels in brain and spinal cord tissues of NSC-transplanted Twi mice, indicating widespread and efficient transport of the bioactive enzyme in CNS tissues (mainly through CSF flow) coupled to active cross-correction. The metabolic correction correlated with amelioration of pathology, clearance of tissue storage and partial rescue of the phenotype. We are currently evaluating the potential therapeutic advantage of GALC-overexpressing NSC as compared to the wt counterpart.Conclusion: These results, together with our preliminary data indicating the feasibility of safe GALC-overexpression in human fetal NSC (a therapeutically relevant cell type), warrant further consideration of NSC gene therapy for the treatment of GLD, likely in combination with other approaches (i.e. bone marrow transplant, currently under evaluation in our laboratory) ensuring enzymatic reconstitution in visceral organs and in the PNS.

Background and Objective: Neurotrophic factors are diffusible polypeptides that have critical roles in survival, proliferation and differentiation of stem cells. This study was done to assess the role of neurotrophic factors (CNTF, BDNF, GDN F, NT- 3) expression and proliferation rate of neural stem cells (NSCS) in coculture with mesenchymal stem cells (MSCs). Materials and Methods: In this experimental study, NSCS and MSCs were isolated from adult Wistar rat. Initially, NSCS was harvested from temporal lobe after mechanical digestion by a sterile flamed Pasteur pipette and enzymatic digestion with trypsin and Dnase. The cell suspension was cultivated in a flask with DMEM/F12 medium supplemented with 10% FBS 100U/ml Penicillin and 100 mg/ml Streptomycin. To obtain MSCs, bone marrow of femur and tibia bones were flashed out and cultured. MSCs and NSCS cocultured by transwell system in DMEM/F12 medium supplemented with 10% FBS 100U/ml Penicillin and 100 mg/ml Streptomycin. Haemocytometer, immunocytochemistry and RT-PCR methods were performed to identify and evaluate cell proliferation, purity levels and neurotrophic factors expression.Results: There is no differences in NTFs profile of neurotrophic factors expression between coculture group and control NSCS, but interactions between MSCs and NSCS significantly promoted NSCS proliferation (P<0.05).Conclusion: This study showed that coculture of NSCS with MSCs might be prfered in cell-therapy than NSCS.

Background and Objectives Uncontrolled stress affects hippocampal-dependent memory through altering the morphology and the proliferation rate of hippocampal progenitor cells. Subjects and Methods In this experimental study, neural stem cells (NSCS) were isolated from the hippocampus of newborn rats and cultivated in a serum-free medium to generate neurosphere. To confirm the induced NSCS, immunocytochemistry and antibody nestin were used. The rate of cell proliferation and cytotoxicity caused by norepinephrine were measured by the MTT assay. NSCS were assigned in the following groups: Control (untreated NSCS), norepinephrine (NSCS treated with norepinephrine), propranolol (NSCS treated with beta receptor blocker propranolol plus norepinephrine), prazosin (NSCS treated with Alfa receptor blocker prazosin plus norepinephrine), and propranolol/ prazosin (NSCS treated with both propranolol and prazosin plus norepinephrine). Real-time PCR was conducted to measure the expression levels of Stk4, Caspase-3 and Sox2 genes. Results The flow cytometry study revealed that NSCS were nestin positive. Real-time PCR results showed that the expression level of Sox2 gene increased by norepinephrine. In addition, the expression level of Stk4 and Caspase-3 genes increased in the groups treated with prazosin. Conclusion The effect of norepinephrine on hippocampus-derived NSCS is receptor-dependent. The increase of norepinephrine under chronic stress can lead to either cell proliferation or apoptosis in NSCS,it acts as a double-edged sword.