Implantation of human urine stem cells promotes neural repair in spinal cord injury rats associated cadeharin-1 and integrin subunit beta 1 expression.

BACKGROUND: The aim of this study was to determine the effect of human urine-derived stem cells (HUSCs) for the treatment of spinal cord injury (SCI) and investigate associated the molecular network mechanism by using bioinformatics combined with experimental validation. METHODS: After the contusive SCI model was established, the HUSC-expressed specific antigen marker was implanted into the injury site immediately, and the Basso, Beattie and Bresnahan locomotor rating scale (BBB scale) was utilized to evaluate motor function so as to determine the effect of HUSCs for the neural repair after SCI. Then, the geneCards database was used to collect related gene targets for both HUSCs and SCI, and cross genes were merged with the findings of PubMed screen. Subsequently, protein-protein interaction (PPI) network, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment, as well as core network construction, were performed using Cytoscape software. Lastly, real-time quantitative polymerase chain reaction (PCR) and immunofluorescence were employed to validate the mRNA expression and localization of 10 hub genes, and two of the most important, designated as cadherin 1 (CDH1) and integrin subunit beta 1 (ITGB1), were identified successfully. RESULTS: The immunophenotypes of HUSCs were marked by CD90+ and CD44+ but not CD45, and flow cytometry confirmed their character. The expression rates of CD90, CD73, CD44 and CD105 in HUSCs were 99.49, 99.77, 99.82 and 99.51%, respectively, while the expression rates of CD43, CD45, CD11b and HLA-DR were 0.08, 0.30, 1.34 and 0.02%, respectively. After SCI, all rats appeared to have severe motor dysfunction, but the BBB score was increased in HUSC-transplanted rats compared with control rats at 28 days. By using bioinformatics, we obtained 6668 targets for SCI and 1095 targets for HUSCs and identified a total of 645 cross targets between HUSCs and SCI. Based on the PPI and Cytoscape analysis, CD44, ACTB, FN1, ITGB1, HSPA8, CDH1, ALB, HSP90AA1 and GAPDH were identified as possible therapeutic targets. Enrichment analysis revealed that the involved signal pathways included complement and coagulation cascades, lysosome, systemic lupus erythematosus, etc. Lastly, quantificational real-time (qRT)-PCR confirmed the mRNA differential expression of CDH1/ITGB1 after HUSC therapy, and glial fibrillary acidic protein (GFAP) immunofluorescence staining showed that the astrocyte proliferation at the injured site could be reduced significantly after HUSC treatment. CONCLUSIONS: We validated that HUSC implantation is effective for the treatment of SCI, and the underlying mechanisms associated with the multiple molecular network. Of these, CDH1 and ITGB1 may be considered as important candidate targets. Those findings therefore provided the crucial evidence for the potential use of HUSCs in SCI treatment in future clinic trials.

A novel cell-free intrathecal approach with PRP for the treatment of spinal cord multiple sclerosis in cats.

BACKGROUND: Multiple sclerosis (MS) is a progressive autoimmune demyelinating disease of the central nervous system. To date, there is no effective therapy for it. Our study aimed to determine the potential role of platelet-rich plasma (PRP) in the treatment of MS in cats. METHODS: The current study was conducted on 15 adult Persian cats that were divided into three groups: control negative, control positive (ethidium bromide (EB)-treated group), and PRP co-treated group (EB-treated group intrathecally injected with PRP on day 14 post-spinal cord injury). PRP was obtained by centrifuging blood on anticoagulant citrate dextrose and activating it with red and green laser diodes. The Basso-Beattie-Bresnahan (BBB) scores were used to assess the motor function recovery on days 1, 3, 7, 14, 20, and 28 following 14 days from EB injection. Moreover, magnetic resonance imaging (MRI) analysis, histopathological investigations, transmission electron microscopy (TEM) studies, and immunohistochemical analysis were conducted, and the gene expressions of nerve growth factors (NGFs), brain-derived neurotrophic factors (BDNF), and stromal cell-derived factors (SDF) were evaluated. RESULTS: Our results indicated that PRP had a significant ameliorative effect on the motor function of the hindlimbs as early as day 20 and so on. MRI revealed that the size and intensity of the lesion were significantly reduced in the PRP co-treated group. The histopathological and TEM investigations demonstrated that the PRP co-treated group had a significant improvement in the structure and organization of the white matter, as well as a high remyelination capacity. Furthermore, a significant increase in myelin basic protein and Olig2 immunoreactivity as well as a reduction in Bax and glial fibrillar acidic protein immune markers was observed. NGFs were found to be upregulated by gene expression. CONCLUSION: As a result, we concluded that the intrathecal injection of PRP was an effective, safe, and promising method for the treatment of MS.

Early mobilization in spinal cord injury promotes changes in microglial dynamics and recovery of motor function.

In the acute phase of spinal cord injury, the initial injury triggers secondary damage due to neuroinflammation, leading to the formation of cavities and glial scars that impair nerve regeneration. Following injuries to the central nervous system, early mobilization promotes the recovery of physical function. Therefore, in the present study, we investigated the effects of early mobilization on motor function recovery and neuroinflammation in rats. Early mobilization of rats with complete spinal cord transection resulted in good recovery of hindlimb motor function after 3 weeks. At 1 week after spinal cord injury, the early-mobilized rats expressed fewer inflammatory M1 microglia/macrophages and more anti-inflammatory M2 microglia. In addition, significantly more matrix metalloproteinase 2 (MMP2)-positive cells were observed at the lesion site 1 week after injury in the early-mobilized rats. Multiple labeling studies suggested that many MMP2-positive cells were M2 microglia. MMP9-positive cells that highly co-expressed GFAP were also observed more frequently in the early-mobilized rats. The density of growth-associated protein-positive structures in the lesion center was significantly higher in the early-mobilized rats at 3 weeks after spinal cord injury. The present results suggest that early mobilization after spinal cord injury reduced the production of M1 microglia/macrophages while increasing the production of M2 microglia at the lesion site. Early mobilization might also activate the expression of MMP2 in M2 microglia and MMP9 in astrocytes. These cellular dynamics might suppress neuroinflammation at the lesion site, thereby inhibiting the progression of tissue destruction and promoting nerve regeneration to recover motor function.

Effect of chronic spinal cord injury's severity on sperm parameters in rat: correlation with locomotion deficits.

OBJECTIVES: Infertility is one of the major concerns for male patients following spinal cord injury (SCI). Although the severity of the injury has a large impact on extent of infertility, the effect of exact injury extent (with specific affected spinal tracts) on fertility is not studied yet. MATERIALS AND METHODS: In the present study, sperm parameters, locomotion scores, and hormonal changes were evaluated following dorsal one third SCI (1/3 SCI), dorsal two third SCI (2/3 SCI), and complete spinal cord transection (TX) at T8 spinal level in male rats. RESULTS: Sperm count decreased significantly following 1/3 SCI and Tx compered to normal (control and sham). In addition, sperm count decreased significantly in Tx compared to 1/3 SCI and 2/3 SCI. Concerning sperm motility, although, percentage of motile sperms decreased significantly in Tx group in comparison to all other groups, the percentage of rapid progressive motile sperms (RPM) decreased significantly in all SCI groups compared to normal. Meanwhile, locomotion score (BBB-score) showed a significant progressive decrease following SCI compared to normal or within SCI groups. However, there was no significant changes in the serum hormonal and seminal fructose concentrations following SCI compared to normal. CONCLUSIONS: These results show that understanding the extent of SCI, the affected spinal tracts, and the resultant locomotion deficits may help to predict the deficits in sperm parameters and hence fertility potentials.

Tailored Modeling of Rivastigmine Derivatives as Dual Acetylcholinesterase and Butyrylcholinesterase Inhibitors for Alzheimer's Disease Treatment.

Rational modification of known drug candidates to design more potent ones using computational methods has found application in drug design, development, and discovery. Herein, we integrate computational and theoretical methodologies to unveil rivastigmine derivatives as dual inhibitors of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) for Alzheimer's disease (AD) management. The investigation entails pharmacokinetics screening, density functional theory (DFT) mechanistic study, molecular docking, and molecular dynamics (MD) simulation. We designed over 20 rivastigmine substituents, subject them to some analyses, and identified RL2 with an appreciable blood-brain barrier score and no permeability glycoprotein binding. The compound shows higher acylation energy and a favored binding affinity to the cholinesterase enzymes. RL2 interacts with the AChE and BuChE active sites showing values of -41.1/-39.5 kcal mol-1 while rivastigmine binds with -32.7/-30.7 kcal mol-1 for these enzymes. The study revealed RL2 (4-fluorophenyl rivastigmine) as a potential dual inhibitor for AChE and BuChE towards Alzheimer's disorder management.

Refinement of the spinal cord injury rat model and validation of its applicability as a model for memory loss and chronic pain.

BACKGROUND: Laminectomy produces trauma in spinal cord injury (SCI) animal models resulting in impinging artefacts and welfare issues. Mechanizing laminectomy using a dental burr assisted (DBA) technique to reduce the impact of conventionally performed laminectomy on animal welfare without any alterations in the outcome of the model was previously demonstrated. However, further validation was necessary to establish it as an alternative in developing SCI rats as a model of chronic pain and memory loss. NOVEL METHOD: DBA technique was employed to perform laminectomy at T10-T11 vertebrae in rats undergoing contusion SCI as a model of chronic pain and memory loss. In a 56-day study, 24 female Wistar rats (Crl: WI) were assigned randomly to four equal groups: conventionally laminectomised, DBA laminectomised, conventionally laminectomised with SCI and DBA laminectomised with SCI. RESULTS: The study revealed DBA technique to cause less surgical bleeding (p = 0.001), lower Rat Grimace Scale (p = 0.0006); resulted in better body weight changes (p = 0.0002 on Day 7 and p = 0.0108 on Day 28) and dark phase activity (p = .0.0014 on Day 1; p = 0.0422 on Day 56). Different techniques did not differ in Basso Beattie Bresnahan score, novel object recognition, mechanical allodynia, number of surviving neurons and the area of vacuolation- indicating that the new method doesn't affect the validity of the model. COMPARISON WITH EXISTING METHODS: In comparison with the conventional technique, motorised laminectomy can be a valid tool that evokes lesser pain and ensures higher well-being in rats modelled for chronic pain and memory loss. CONCLUSIONS: The intended outcome from the model is not influenced by techniques whereas the DBA-technique is a refined alternative to the conventional method in achieving better welfare in SCI studies.

The Comparative Effects of Mesenchymal Stem Cell Transplantation Therapy for Spinal Cord Injury in Humans and Animal Models: A Systematic Review and Meta-Analysis.

Animal models have been used in preclinical research to examine potential new treatments for spinal cord injury (SCI), including mesenchymal stem cell (MSC) transplantation. MSC transplants have been studied in early human trials. Whether the animal models represent the human studies is unclear. This systematic review and meta-analysis has examined the effects of MSC transplants in human and animal studies. Following searches of PubMed, Clinical Trials and the Cochrane Library, published papers were screened, and data were extracted and analysed. MSC transplantation was associated with significantly improved motor and sensory function in humans, and significantly increased locomotor function in animals. However, there are discrepancies between the studies of human participants and animal models, including timing of MSC transplant post-injury and source of MSCs. Additionally, difficulty in the comparison of functional outcome measures across species limits the predictive nature of the animal research. These findings have been summarised, and recommendations for further research are discussed to better enable the translation of animal models to MSC-based human clinical therapy.

Stem cell treatment trials of spinal cord injuries in animals.

BACKGROUND: Spinal cord injury (SCI) is a serious neurological spinal cord damage that resulted in the loss of temporary or permanent function. However, there are even now no effective therapies for it. So, a new medical promising therapeutic hotspot over the previous decades appeared which was (Stem cell (SC) cure of SCI). Otherwise, animal models are considered in preclinical research as a model for humans to trial a potential new treatment. METHODOLOGY: Following articles were saved from different databases (PubMed, Google scholar, Egyptian knowledge bank, Elsevier, Medline, Embase, ProQuest, BMC) on the last two decades, and data were obtained then analyzed. RESULTS: This review discusses the type and grading of SCI. As well as different types of stem cells therapy for SCI, including mesenchymal stem cells (MSCs), neural stem cells (NSCs), hematopoietic stem cells (HSCs), induced pluripotent stem cells (iPSCs), and embryonic stem cells (ESCs). The review focuses on the transplantation pathways, clinical evaluation, and clinical signs of different types of SC on different animal models which are summarized in tables to give an easy to reach. CONCLUSION: Pharmacological and physiotherapy have limited regenerative power in comparison with stem cells medication in the treatment of SCI. Among several sources of cell therapies, mesenchymal stromal/stem cell (MSC) one is being progressively developed as a trusted important energetic way to repair and regenerate. Finally, a wide-ranged animal models have been condensed that helped in human clinical trial therapies.

Effect of Fingomod on the Inflammatory Response of Cerebral Ischemia-reperfusion Injury in Rats / 中风与神经疾病杂志

@#Objective To investigate the effect of Fingomod（FTY720）on the cerebral ischemia-reperfusion injury in rats. Methods SPF rats（250 g～300 g）of 8 weeks old were divided into sham operation group，blank control group，group of Clopidogrel and group of FTY720.Rat model of ischemic stroke was made by tMCAO. The behavioral disorder scores were scored in different groups after different drug treatments. And observing the area of infarctus tissue by the TTC method and the nerve cells changed by HE method in different groups. The levels of inflammatory factors IL-1β and IL-6 in brain tissue of rats were changed by method of ELISA. The level of p-p38MAPK and NF-κB in brain tissue of rats were determined by Western-blot method. Results（1）FTY720 can alleviate the behavior disorder of rats（P<0.01）；（2）FTY720 can reduce the area of infarctus tissue（P<0.01）and the injury of neurology cells in rats；（3）FTY720 can reduce the level of p-p38MAPK and NF-κB（P<0.01）in the ischemic brain tissue of rats；4，FTY720 can be reduced The level of IL-1β（P<0.01）and IL-6（P<0.01）in the ischemic brain tissue. Conclusion Fingomod（FTY720）has a protective effect on the cerebral ischemia-reperfusion rat model by decreasing the level of inflammatory response.

The time window of osimertinib in the treatment of spinal cord injury in rats / 中风与神经疾病杂志

@#Objective　To determine the optimal time window to treat spinal cord injury (SCI) by epidermal growth factor receptor (EGFR) inhibitor osimertinib.Methods　60 rats were randomly divided into 6 groups (10 in each group):injury control group and 5 osimertinib-treated groups (0 d,1 d,3 d，5 d and 7 d group).After rats spinal cord injury model was established,the rats in osimertinib-treated group were applied with at different time (0 day,1 day,3 days and 7 days) after injury，while the injury group animals were only given vehicle solution (normal saline) lacking osimertinib.Neurological deficits was evaluated by using the BBB locomotor scale at first day and weekly post-injury.Residual urine volume was recorded everyday in each group.14 days after SCI, 5 rats were randomly selected and sacrificed.Spinal cord tissues were collected,and the expression of GAP-43 was detected by Western blot.Results　(1)The BBB score of rats in 0 d group,1 d group,3 d group was significantly higher than 5 d group,7 d group and injury groups(P<0.05),and there was no significant difference among 0 d group,1 d group and 3 d group(P>0.05).(2)The residual urine volume of rats in 0 d group,1 d group,3 d group was significantly less than that other groups (P<0.05),and there was no significant difference among 0 d group,1 d group and 3 d group(P>0.05).(3)The expression of GAP-43 in 0 d group,1 d group and 3 d group was significantly higher than other groups(P<0.05,or P<0.01),and there was no significant difference among 0 d group,1 d group and 3 d group (P>0.05).Conclusion　The optimal time of EGFR inhibitor osimertinib to treat spinal cord injury in rats was 0 h to 72 h after spinal cord injury.

Improved spinal cord gray matter morphology induced by Spirulina platensis following spinal cord injury in rat models.

Despite intense preclinical research focusing on developing potential strategies of mitigating spinal cord injury (SCI), SCI still results in permanent, debilitating symptoms for which there are currently no effective pharmacological interventions to improve the recovery of the fine ultrastructure of the spinal cord. Spirulina platensis is thought to have potential neuroprotective effects. We have previously demonstrated its protective potential on the lesioned corticospinal tracts and behavioral recovery. In this study, spirulina, known for its neuroprotective properties was used to further explore its protective effects on spinal cord gray matter ultrastructural. Twenty-four Sprague-Dawley rats were used and divided into sham group (laminectomy without SCI), control group (SCI without S. platensis), and S. platensis group (SCI + 180 mg/kg S. platensis). All animals were anesthetized via intramuscular injection. A partial crush injury was induced at the level of T12. The rats were humanely sacrificed for 28 days postinjury for ultrastructural study. There were significant mean differences with respect to pairwise comparisons between the ultrastructural grading score of neuronal perikarya of control and the S. platensis following injury at day 28, which correlates with the functional locomotor recovery at this timepoint in our previous study. The group supplemented with spirulina, thus, revealed a better improvement in the fine ultrastructure of the spinal cord gray matter when compared to the control group thereby suggesting neuroprotective potentials of spirulina in mitigating the effects of spinal cord injury and inducing functional recovery.

Characterization of transection spinal cord injuries by monitoring somatosensory evoked potentials and motor behavior.

Standardization of spinal cord injury (SCI) models is crucial for reproducible injury in research settings and their objective assessments. Basso, Beattie and Bresnahan (BBB) scoring, the traditional behavioral evaluation method, is subjective and susceptible to human error. On the other hand, neuro-electrophysiological monitoring, such as somatosensory evoked potential (SSEP), is an objective assessment method that can be performed continuously for longitudinal studies. We implemented both SSEP and BBB assessments on transection SCI model. Five experimental groups are designed as follows: left hemi-transection at T8, right hemi-transection at T10, double hemi-transection at left T8 and right T10, complete transection at T8 and control group which receives only laminectomy with intact dura and no injury on spinal cord parenchyma. On days 4, 7, 14 and 21 post-injury, first BBB scores in awake and then SSEP signals in anesthetized rats were obtained. Our results show SSEP signals and BBB scores are both closely associated with transection model and injury progression. However, the two assessment modalities demonstrate different sensitivity in measuring injury progression when it comes to late-stage double hemi-transection, complete transection and hemi-transection injury. Furthermore, SSEP amplitudes are found to be distinct in different injury groups and the progress of their attenuation is increasingly rapid with more severe transection injuries. It is evident from our findings that SSEP and BBB methods provide distinctive and valuable information and could be complementary of each other. We propose incorporating both SSEP monitoring and conventional BBB scoring in SCI research to more effectively standardize injury progression.

In vitro differentiation potential of human urine-derived stem cells into neuron-like cells and their protective effects against spinal cord injury in rats / 中国组织工程研究

BACKGROUND: Human urine-derived stem cells are newly discovered adult stem cells, characterized by rich sources, simple extraction, good proliferative ability and multi-directional differentiation potential. In recent years, human urine-derived stem cells have been used for the repair of neurological functions in urinary diseases, such as stress urinary incontinence and vesicoureteral reflux. OBJECTIVE: To explore the biological characteristics of human urine-derived stem cells and to study their repairing effect in a rat model of spinal cord injury. METHODS: Cell phenotypes of human urine-derived stem cells were detected using flow cytometry, and the immunohistochemical staining was used to identify neuron-like cells differentiated from human urine-derived mesenchymal stem cells. Then, an animal model of spinal cord injury at T9 segment was made by Allen method, and after modeling 24 Sprague-Dawley rats were assigned into spinal cord injury group or cell treatment group (n=12/group). In the cell treatment group, the model rats were injected 2 μL of 1.0×1011/L human urine-derived stem cells, while in the spinal cord injury group, the rats were administered the same volume of L-DMEM containing 10% fetal bovine serum. Basso, Beattie and Bresnahan scores were valued at 1, 10, 20, and 30 days after modeling. Spinal cord samples from all the rats were taken out at 30 days after modeling, and Luxol Fast Blue staining, microglia/macrophages staining and glial fibrillary acidic protein staining were used to value the injured area of the spinal cord and the fluorescence intensity of glial fibrillary acidic protein. RESULTS AND CONCLUSION: (1) Flow cytometry showed high expression on CD29 and CD90, and low expression on CD45 in human urine-derived mesenchymal stem cells. Moreover, human urine-derived mesenchymal stem cells could be induced to differentiating into neuron-like cells in vitro. (2) Basso, Beattie and Bresnahan scores showed no significant difference between the two groups at 1 and 10 days after modeling (P > 0.05), while, at 20 and 30 days after modeling, the scores in the cell treatment group were significantly higher than those in the spinal cord injury group (P < 0.05). (3) Luxol Fast Blue staining showed that the injured area of the spinal cord in the cell treatment group was markedly less than that in the spinal cord injury group (P < 0.05), and the glial fibrillary acidic protein showed lower fluorescence intensity in the cell treatment group than the spinal cord injury group (P < 0.05). To conclude, human urine-derived stem cells can differentiate into neuron-like cells and have therapeutic effects in the rat model of spinal cord injury.

Neuroprotective potential of Spirulina platensis on lesioned spinal cord corticospinal tract under experimental conditions in rat models.

Spinal cord injury (SCI) results from penetrating or compressive traumatic injury to the spine in humans or by the surgical compression of the spinal cord in experimental animals. In this study, the neuroprotective potential of Spirulina platensis was investigated on ultrastructural and functional recovery of the spinal cord following surgical-induced injury. Twenty-four Sprague-Dawley rats were divided into three groups; sham group, control (trauma) group, and experimental (S. platensis) group (180 mg/kg) of eight rats each. For each group, the rats were then subdivided into two groups to allow measurement at two different timepoints (day 14 and 28) for the microscopic analysis. Rats in the control and experimental S. platensis groups were subjected to partial crush injury at the level of T12 with Inox number 2 modified forceps by compressing on the spinal cord for 30 s. Pairwise comparisons of ultrastructural grading mean scores difference between the control and experimental S. platensis groups reveals that there were significant differences on the axonal ultrastructure, myelin sheath and BBB Score on Day 28; these correlate with the functional locomotor recovery at this timepoint. The results suggest that supplementation with S. platensis induces functional recovery and effective preservation of the spinal cord ultrastructure after SCI. These findings will open new potential avenue for further research into the mechanism of S. platensis-mediated spinal cord repair.

Ursodeoxycholic Acid Inhibits Inflammatory Responses and Promotes Functional Recovery After Spinal Cord Injury in Rats.

The aim of this study was to investigate the anti-inflammatory effects by ursodeoxycholic acid (UDCA) in rats with a spinal cord injury (SCI). A moderate mechanical compression injury was imposed on adult Sprague-Dawley (SD) rats. The post-injury locomotor functions were assessed using the Basso, Beattie, and Bresnahan (BBB) locomotor scale and the tissue volume of the injured region was analyzed using hematoxylin and eosin staining. The pro-inflammatory factors were evaluated by immunofluorescence (IF) staining, a quantitative real-time polymerase chain reaction (qRT-PCR), and enzyme-linked immunosorbent assay (ELISA). The phosphorylation of the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 in mitogen-activated protein kinase (MAPK) signaling pathways related to inflammatory responses were measured by Western blot assays. UDCA improved the BBB scores and promoted the recovery of the spinal cord lesions. UDCA inhibited the expression of glial fibrillary acidic protein (GFAP), tumor necrosis factor-α (TNF-α), ionized calcium-binding adapter molecule 1 (iba1), and inducible nitric oxide synthase (iNOS). UDCA decreased the pro-inflammatory cytokines of TNF-α, interleukin 1-ß (IL-1ß), and interleukin 6 (IL-6) in the mRNA and protein levels. UDCA increased the anti-inflammatory cytokine interleukin 10 (IL-10) in the mRNA and protein levels. UDCA suppressed the phosphorylation of ERK, JNK, and the p38 signals. UDCA reduces pro-inflammatory responses and promotes functional recovery in SCI in rats. These results suggest that UDCA is a potential therapeutic drug for SCI.

Induced Pluripotent Stem Cell Transplantation Improves Locomotor Recovery in Rat Models of Spinal Cord Injury: a Systematic Review and Meta-Analysis of Randomized Controlled Trials.

BACKGROUND/AIMS: Spinal cord injury (SCI) has long been a subject of great interest in a wide range of scientific fields. Several attempts have been made to demonstrate motor function improvement in rats with SCI after transplantation of induced pluripotent stem cells (iPSC). This systematic review and meta-analysis was designed to summarize the effects of iPSC on locomotor recovery in rat models of SCI. METHODS: We searched the publications in the PubMed, Medline, Science Citation Index, Cochrane Library, CNKI, and Wan-fang databases and the China Biology Medicine disc. Results were analyzed by Review Manager 5.3.0. The quality of evidence was assessed using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology. RESULTS: Six randomized controlled preclinical trials covering eight comparisons and including 212 rats were selected. The subgroup analyses were based on the following items: different SCI models, cell counts, iPSC sources, iPSC differentiations and transplantation methods. The pooled results indicated that iPSC transplantation significantly improved locomotor recovery of rats after SCI by sustaining beneficial effects, especially in the subgroups of contusion, moderate cell counts (5×105), source of human fetal lung fibroblasts, iPSC-neural precursors and intraspinal injection. CONCLUSION: Our meta-analysis of the effects of iPSC transplantation on locomotor function in SCI models is, to our knowledge, the first meta-analysis in this field. We conclude that iPSC transplantation improves locomotor recovery in rats with SCI, implicating this strategy as an effective therapy. However, more studies are required to validate our conclusions.

miR-411 suppresses acute spinal cord injury via downregulation of Fas ligand in rats.

OBJECTIVE: To explore the role of miR-411/FasL in acute spinal cord injury (ASCI). METHODS: The ASCI rat model was established, and expression of miR-411 and Fas ligand (FasL) was examined. Basso, Beattie and Bresnahan (BBB) score was used to evaluate the rats' neurological function. PC12 oxygen-glucose deprivation (OGD) model was also established. Gene manipulation (including miR-411 mimic or inhibitor) was used to modulate gene expression. Luciferase reporter assay was conducted to confirm the targeting relationship between miR-411 and FasL. Flow cytometry was applied in the measurement of PC12 cell apoptosis. Finally, the miR-411 mimic was injected into the vertebral canal of ASCI rats to determine the effects of miR-411 in vivo. RESULTS: Compared with sham group, the expression of miR-411 and FasL was significantly decreased and increased in ASCI group, respectively (P < 0.05). Similarly, the expression of miR-411 and FasL was significantly lower and higher in OGD group than that in control group, respectively (P < 0.05). miR-411 directly controlled the FasL expression. miR-411 mimic can dramatically reduce the increased percentage of apoptosis cells caused by OGD when comparing to mimic control, which was greatly reversed by the overexpression of FasL (P < 0.05). Further, the BBB score was significantly elevated in the miR-411 mimic group when comparing to mimic control group, with decreased FasL expression (P < 0.05). CONCLUSION: miR-411 mimic suppressed PC12 cell apoptosis via FasL, and relieved ASCI in rats.

Multilayer polyion complex nanoformulations of superoxide dismutase 1 for acute spinal cord injury.

As one of the most devastating forms of trauma, spinal cord injury (SCI) remains a challenging clinical problem. The secondary processes associated with the primary injury, such as overproduction of reactive oxygen species (ROS) and inflammation, lead to concomitant compression of the injured spinal cord and neuronal death. Delivery of copper-zinc superoxide dismutase (SOD1), an efficient ROS scavenger, to the site of injury can mitigate SCI-induced oxidative stress and tissue damage. Towards this goal catalytically active nanoformulations of SOD1 ("nanozymes") are developed as a modality for treatment of SCI. Along with the cross-linked polyion complex of SOD1 with polycation poly(ethylene glycol) (PEG)-polylysine (single-coat (SC) nanozyme), we introduce for the first time the chemically cross-linked multilayer polyion complex in which SOD1 is first incorporated into a polyion complex with polycation, then coated by anionic block copolymer, PEG-polyglutamic acid (double-coat (DC) nanozyme). We developed DC nanozymes with high enzymatic activity and ability to retain and protect SOD1 under physiological conditions. Pharmacokinetic study revealed that DC nanozymes significantly prolonged circulation of active SOD1 in the blood stream compared to free SOD1 or SC nanozymes (half-life was 60 vs 6min). Single intravenous injection of DC nanozymes (5kU of SOD1/kg) improved the recovery of locomotor functions in rats with moderate SCI, along with reduction of swelling, concomitant compression of the spinal cord and formation of post-traumatic cysts. Thus, based on the testing in a rodent model the SOD1 DC nanozymes are promising modality for scavenging ROS, decreasing inflammation and edema, and improving recovery after SCI.

Varied therapeutic effects of Human Amniotic Mesenchymal Stem Cell Transplantation with Different Transplantation Methods for Spinal Cord Injury Therapy in Rats / 实用医学杂志

Objective To compare the therapeutic effects of orthotopic injection and tail vein injection of human amniotic mesenchymal stem cells(hAMSCs)on histological restoration and neurological functions of rats with spinal cord injury. Methods Transected spinal cord injury model in rats was established by transplanting DAPI prelabelled hAMSCs one week after injury.BBB scores were used to evaluate the hindlimb movement of rats. The histological patterns.and morphology of medullary sheath of spinal cord were observed. Results BBB scores in the orthotopic injection group and tail vein injection group were increased gradually from one to six week after hAM-SCs transplantation and reached 6.5 ± 0.5 and 7.12 ± 1.61 respectively 6 weeks after cell transplantation,higher than that of the control group(both P < 0.01). However,there was no statistical significance between the two groups.Histological results indicated that the repair of injured tissue in the orthotopic injection group and tail vein injection group were both better than that in the control group,and there were more vesica and loosened layers forming in the injured spinal cord of rats in the PBS control group as compared with the orthotopic and tail vein transplantation group. Conclusion hAMSCs transplantation through tail vein injection could promote histological restoration and neurological regeneration of rats with spinal cord injury,which has the similar therapeutic effects with hAMSCs orthotopic transplantation.

Globose basal cells for spinal cord regeneration.

Spinal cord injury (SCI) is a devastating condition with loss of motor and sensory functions below the injury level. Cell based therapies are experimented in pre-clinical studies around the world. Neural stem cells are located intra-cranially in subventricular zone and hippocampus which are highly invasive sources. The olfactory epithelium is a neurogenic tissue where neurogenesis takes place throughout the adult life by a population of stem/progenitor cells. Easily accessible olfactory neuroepithelial stem/progenitor cells are an attractive cell source for transplantation in SCI. Globose basal cells (GBCs) were isolated from rat olfactory epithelium, characterized by flow cytometry and immunohistochemically. These cells were further studied for neurosphere formation and neuronal induction. T10 laminectomy was done to create drop-weight SCI in rats. On the 9th day following SCI, 5 × 105 cells were transplanted into injured rat spinal cord. The outcome of transplantation was assessed by the Basso, Beattie and Bresnahan (BBB) locomotor rating scale, motor evoked potential and histological observation. GBCs expressed neural stem cell markers nestin, SOX2, NCAM and also mesenchymal stem cell markers (CD29, CD54, CD90, CD73, CD105). These cells formed neurosphere, a culture characteristics of NSCs and on induction, differentiated cells expressed neuronal markers ßIII tubulin, microtubule-associated protein 2, neuronal nuclei, and neurofilament. GBCs transplanted rats exhibited hindlimb motor recovery as confirmed by BBB score and gastrocnemius muscle electromyography amplitude was increased compared to controls. Green fluorescent protein labelled GBCs survived around the injury epicenter and differentiated into ßIII tubulin-immunoreactive neuron-like cells. GBCs could be an alternative to NSCs from an accessible source for autologous neurotransplantation after SCI without ethical issues.