Acute Spinal Cord Injury Model Research Articles

Evaluation of the Chetomin effect on histopathological features in a murine acute spinal cord injury model

BackgroundSeveral research studies have been focused on improving the treatment and prognosis of acute spinal cord injury, as part of this initiative we investigated the use of Chetomin to reduce the inflammatory response in this pathology. MethodsAn experimental, prospective, cross-sectional study was performed using 42 Wistar rats where we analyzed the effect of Chetomin compared to methylprednisolone administered 1 and 8 h after the spinal cord injury in a murine model. ResultsChetomin administration 8h post-injury decreased IL-6 and VEGF expression; and, and its administration 1h post-injury decreased NF-kB expression. ConclusionsChetomin has anti-inflammatory effects in acute spinal cord injury, whether these effects are observable with other proinflammatory markers should be investigated.

Targeted-delivery of nanomedicine-enabled methylprednisolone to injured spinal cord promotes neuroprotection and functional recovery after acute spinal cord injury in rats

To date, no therapy has been proven to be efficacious in fully restoring neurological functions after spinal cord injury (SCI). Systemic high-dose methylprednisolone (MP) improves neurological recovery after acute SCI in both animal and human. MP therapy remains controversial due to its modest effect on functional recovery and significant adverse effects. To overcome the limitation of MP therapy, we have developed a N-(2-hydroxypropyl) methacrylamide copolymer-based MP prodrug nanomedicine (Nano-MP) that can selectively deliver MP to the SCI lesion when administered systemically in a rat model of acute SCI. Our in vivo data reveal that Nano-MP is significantly more effective than free MP in attenuating secondary injuries and neuronal apoptosis. Nano-MP is superior to free MP in improving functional recovery after acute SCI in rats. These data support Nano-MP as a promising neurotherapeutic candidate, which may provide potent neuroprotection and accelerate functional recovery with improved safety for patients with acute SCI.

Mechanism of Shikonin on spinal cord injury in rats based on TNFR/RIPK1 pathway

To explore the effect of shikonin on the recovery of nerve function after acute spinal cord injury(SCI) in rats. 96 male Sprague-Dawley(SD)rats were divided into 4 groups randomly:sham operation group (Group A), sham operation+shikonin group (Group B), SCI+ DMSO(Group C), SCI+shikonin group (Group D).The acute SCI model of rats was made by clamp method in groups C and D . After subdural catheterization, no drug was given in group A. rats in groups B and D were injected with 100 mg·kg-1 of shikonin through catheter 30 min after modeling, and rats in group C were given with the same amount of DMSO, once a day until the time point of collection tissue. Basso-Beattie-Bresnahan(BBB) scores were performed on 8 rats in each group at 6, 12, and 3 d after moneling, and oblique plate tests were performed on 1, 3, 7 and 14 d after modeling, and then spinal cord tissues were collected. Eight rats were intraperitoneally injected with propidine iodide(PI) 1 h before sacrificed to detection PI positive cells at 24 h in each group. Eight rats were sacrificed in each group at 24 h after modeling, the spinal cord injury was observed by HE staining.The Nissl staining was used to observe survivor number of nerve cells. Western-blot technique was used to detect the expression levels of Bcl-2 protein and apoptosis related protein RIPK1. After modeling, BBB scores were normal in group A and B, but in group C and D were significantly higher than those in group A and B. And the scores in group D were higher than those in group C in each time point (P<0.05). At 12 h after modeling, the PI red stained cells in group D were significantly reduced compared with that in group C, and the disintegration of neurons was alleviated(P<0.05). HE and Nissl staining showed nerve cells with normal morphology in group A and B at 24h after operation. The degree of SCI and the number of neuronal survival in group D were better than those in group C, the difference was statistically significant at 24h (P<0.05). The expression of Bcl-2 and RIPK1 proteins was very low in group A and B;The expression of RIPK1 was significantly increased in Group C and decreased in Group D, with a statistically significant difference (P<0.05);The expression of Bcl-2 protein in group D was significantly higher than that in group C (P<0.05). Shikonin can alleviate the pathological changes after acute SCI in rats, improve the behavioral score, and promote the recovery of spinal nerve function. The specific mechanism may be related to the inhibition of TNFR/RIPK1 signaling pathway mediated necrotic apoptosis.

Anti-Siglec-15 Antibody Prevents Marked Bone Loss after Acute Spinal Cord Injury-Induced Immobilization in Rats.

Rapid and extensive sublesional bone loss after spinal cord injury (SCI) is a difficult medical problem that has been refractory to available interventions except the antiresorptive agent denosumab (DMAB). While DMAB has shown some efficacy in inhibiting bone loss, its concurrent inhibition of bone formation limits its use. Sialic acid-binding immunoglobulin-like lectin (Siglec)-15 is expressed on the cell surface of mature osteoclasts. Anti-Siglec-15 antibody (Ab) has been shown to inhibit osteoclast maturation and bone resorption while maintaining osteoblast activity, which is distinct from current antiresorptive agents that inhibit the activity of both osteoclasts and osteoblasts. The goal of the present study is to test a Siglec-15 Ab (NP159) as a new treatment option to prevent bone loss in an acute SCI model. To this end, 4-month-old male Wistar rats underwent complete spinal cord transection and were treated with either vehicle or NP159 at 20 mg/kg once every 2 weeks for 8 weeks. SCI results in significant decreases in bone mineral density (BMD, -18.7%), trabecular bone volume (-43.1%), trabecular connectivity (-59.7%), and bone stiffness (-76.3%) at the distal femur. Treatment with NP159 almost completely prevents the aforementioned deterioration of bone after SCI. Blood and histomorphometric analyses revealed that NP159 is able to greatly inhibit bone resorption while maintaining bone formation after acute SCI. In ex vivo cultures of bone marrow cells, NP159 reduces osteoclastogenesis while increasing osteoblastogenesis. In summary, treatment with NP159 almost fully prevents sublesional loss of BMD and metaphysis trabecular bone volume and preserves bone strength in a rat model of acute SCI. Because of its unique ability to reduce osteoclastogenesis and bone resorption while promoting osteoblastogenesis to maintain bone formation, Siglec-15 Ab may hold greater promise as a therapeutic agent, compared with the exclusively antiresorptive or anabolic agents that are currently used, in mitigating the striking bone loss that occurs after SCI or other conditions associated with severe immobilization. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Effect of ACTH4-10Pro8-Gly9-Pro10 on anti-inflammatory cytokine (IL-4, IL-10, IL-13) expression in acute spinal cord injury models (male Sprague Dawley rats)

Background: Spinal cord injury (SCI) is a damage to the spinal cord caused mainly by trauma resulting in major motor, sensory and autonomic dysfunctions. Its final neurological outcome is determined by both primary and secondary injury processes. A key component of secondary injury mechanisms after initial trauma is neuroinflammation. A neuroprotective compound, ACTH4-10Pro8-Gly9-Pro10 (ACTH4-10) also known as semax, has shown neuroprotective and anti-inflammatory properties. ACTH4-10 has also been actively used in the treatment of brain ischemia without serious complication reported. Here, we analyzed the effects of ACTH4-10 at regulating the inflammatory cascade in SCI by looking at anti-inflammatory cytokine (IL-4, IL-10 and IL-13) levels after acute SCI. Method: We carried out laminectomies in male Sprague Dawley rats at the second thoracic vertebrae. After laminectomy, we exposed the myelum and created mild SCI models with 20-g, and severe SCI with 35-g aneurysm clips. ACTH4-10 was administered intranasally to the treatment group and 0.9% NaCl to the control group (placebo). Both groups were kept alive and terminated at 3 and 6 hours. The tissue sample preparations were fixed in formalin and examined for immunohistochemistry. Quantitative measurement of the cytokines was done in the posterior horn area with specific associated anti-monoclonal antibodies. Results: Rats with mild SCI that were given ACTH4-10 showed greater anti-inflammatory levels at 3 hours post-compression but only IL-10 and IL-13 were elevated significantly at 6 hours. Rats with severe compression in ACTH4-10 group showed greater levels of IL-10, IL-13 at 3 hours and IL-4, IL-10 at 6 hours compared with the placebo group. Conclusions: Administration of ACTH4-10Pro8-Gly9-Pro10 intranasal can increase anti-inflammatory cytokine expression in Sprague Dawley rat models with mild and severe SCI. Expression of anti-inflammatory cytokines was greater in mild compression and 3-hour termination. Further research is needed to determine the optimal dose and clinical outcome in vivo.

Evaluation of the chetomin effect on histopathological features in a murine acute spinal cord injury model

Evaluation of the chetomin effect on histopathological features in a murine acute spinal cord injury model

N-Acetylcysteine alleviates spinal cord injury in rats after early decompression surgery by regulating inflammation and apoptosis

ABSTRACT Objective: Decompression surgery in patients with spinal cord injury (SCI) has a neuroprotective effect by alleviating secondary injury and improving neurological outcomes. N-Acetylcysteine (NAC), a drug approved by the United States Food and Drug Administration, has been shown to play neuroprotective roles via attenuation of apoptosis and inflammation. The purpose of the present study was to investigate the effects of early or late decompression surgery in combination with NAC administration on acute SCI, as well as investigate the underlying mechanisms of its actions. Methods: In this study, an acute SCI model was established in rats. The rats were treated with decompression surgery 24/48 h post-SCI in combination with or without NAC. Results: The results showed that decompression surgery in combination with NAC lead to a better outcome than decompression alone, as demonstrated by the higher Basso, Beattie, and Bresnahan scores. Histopathological examination demonstrated that early decompression surgery in combination with NAC exerted the best therapeutic effect on spinal cord recovery, which was further confirmed by the extent of inflammation and apoptosis. Additionally, we found that NAC might compensate for a lack of late surgery. Conclusions: Collectively, early decompression surgery and NAC could be a promising combination for the treatment of acute SCI, and its therapeutic effects may be associated with the regulation of inflammation and apoptosis.

Association of Olfactory Bulb Transplantation with Systemic Drugs: Paclitaxel (Taxol), Ambroxol and Colchicine in a Mouse Acute Spinal Cord Injury Model: Summatory Synergy

Multiple therapeutics looking for functional motor reconnection in spinal cord injury has been reported with unpromising results; they are not yet acceptable for clinical application. They have been studied individually or in combinations looking for synergy mostly on therapeutic attempts of chronic injury and on few experimental acute injury models.

Downregulation of Nck1 After Spinal Cord Injury in Adult Rats

Background:Nck1 is an important molecule that participates in many cellular processes, including neurite outgrowth, synaptic plasticity, and apoptosis. However, the expression and function of Nck1 in the spinal cord and spinal cord injury remain unknown.Aims:To investigate the role of Nck1 in spinal cord injury.Study Design:Animal experimentation.Methods:Adult Sprague–Dawley rats were used to establish an acute spinal cord injury model. Double immunofluorescence staining, Western blot, and quantitative reverse transcription polymerase chain reaction analysis were used to investigate the distribution, cellular localization, and expression of Nck1 in spinal cord injury processes. Short interfering RNA was used to silence Nck1 expression in VSC4.1 cells. The Shapiro–Wilk test was used for the normality distribution analysis; the Student’s unpaired t-test, 1-way analysis of variance followed by post hoc Tukey’s test were used for data analysis. Finally, RNA sequencing technology and gene ontology analysis were used to analyze the changes in Nck1-associated genes expression after spinal cord injury.Results:Colabeled staining demonstrated that Nck1 was especially distributed in neurons. Western blot, quantitative reverse transcription polymerase chain reaction, and statistical analysis revealed that Nck1 expression reduced to the lowest levels at 1 day after nerve injury, and slowly increased to a stable level in 21 days (P < .05). Nck1-specific short interfering RNA transfection significantly reduced cell viability and neurite development in neurons. Bioinformatic analysis indicated that Nck1 participates in multiple pathological processes of spinal cord injury, and many Nck1-associated genes exhibited differential expression levels.Conclusion:Nck1 is a vital protein in spinal cord injury processes and, therefore, further studies should be conducted to explore its potential functions and molecular mechanisms in spinal cord injury repair.

PU.1 interaction with p50 promotes microglial-mediated inflammation in secondary spinal cord injury in SCI rats

Purpose/aim of the study Secondary spinal cord injury is the inflammatory damage to surrounding tissues caused by activated microglial-mediated neuroinflammatory responses. The nuclear factor-κB (p65/p50) pathway and PU.1 are closely correlated with inflammatory responses; thus, we examined the relationship and function between PU.1 and p50 in secondary spinal cord injury.Materials and methods In this study, we established an adult rat acute spinal cord injury model to simulate the pathological process of spinal cord injury.Results: We found that the expression of PU.1 was significantly increased at three days after spinal cord injury and mainly expressed in activated microglia. Moreover, p-p50 expression was increased in SCI rats and the protein interacted with PU.1. Lipopolysaccharide was used to induce microglia activation in vitro.Conclusions: The results showed that PU.1 and p-p50 expression was significantly increased and PU.1 interacted with p50 in the nucleus. The levels of tumor necrosis factor-α and interleukin-1β secreted by microglia were detected by enzyme-linked immunosorbent assay. The results showed that when both PU.1 and p50 were overexpressed, tumor necrosis factor-α and interleukin-1β secretion was significantly increased to levels higher than in cells overexpressing PU.1 or p50 alone. These results suggest that PU.1 and p50 interact to promote p65 transcription and the expression of inflammatory factors, which is an important mechanism of the microglial-mediated inflammatory response to secondary injury after spinal cord injury.

Influence of electroacupuncture on the expression of AMPA receptor subunit GluR1 in the spinal injured area of the rats with acute spinal cord injury

To explore the influence of electroacupuncture (EA) on the expression of AMPA receptor subunit GluR1 in the rats with acute spinal cord injury (SCI) and explore the potential effect mechanism of EA in treatment of acute SCI. A total of 80 SD rats were randomly divided into five groups, i.e. a sham-operation group, a model group, an AMPA antagonist (DNQX) group, an EA group and a DNQX+EA group, 16 rats in each group. The modified Allen's impacting method was adopted to prepare the rat model of acute SCI at T10. In the DNQX group, the intrathecal injection of 10 μL DNQX solution with a concentration of 1 nmol/μL was administered in 0.5 h after modeling success. In the EA group, EA (disperse-dense wave, 2 Hz/100 Hz in frequency, 0.5 mA in output current) was given at "Dazhui" (GV 14) and "Mingmen" (GV 4) in 0.5 h, 12 h and 24 h after modeling success for 30 min and totally 3 times. In the DNQX + EA group, the interventions in the above two groups were managed. The Basso, Beattie and Bresnahan locomotor rating score (BBB) was applied to evaluate the changes of locomotor function in the rats before modeling and in 6 h, 24 h and 48 h after modeling successively. Using the hematoxylin-eosin (HE) staining, the histopathological changes in the spinal anterior horn were observed in the spinal injured area. The immunofluorescence method was adopted to determine the number of GluR1 positive neuron of the spinal anterior horn. The Western blot method was used to determine the protein expression of GluR1 in the injured area. Compared to the sham-operation group in 6 h, 24 h and 48 h after modeling, the BBB scores were all significantly decreased in the model group (P<0.001) at the corresponding points. The BBB score was increased in each of intervention groups, but without statistical difference as compared with the model group (P>0.05). In the model group, it was found that the boundary between gray matter and white matter in the spinal anterior horn was blurred, the interstitial space enlarged, the neuron volume obviously shrunken, the cytoplasm decreased, the red stain deepened and some neuron nuclei fixed and shrunk. In the EA group, the morphology of the spinal anterior horn in the injured area was improved obviously, which was similar in the DNQX group and the DNQX + EA group. Compared with the sham-operation group, the GluR1 protein expression in the spinal injury area was increased (P<0.001) and the number of GluR1 positive neurons elevated (P<0.001) in the spinal anterior horn in the model group. Compared with the model group, in the EA group, the DNQX group and the DNQX + EA group, GluR1 protein expression was decreased (P<0.05, P<0.01) and the number of GluR1 positive neurons in the spinal anterior horn reduced (P<0.001). The intervention with EA at "Dazhui" and "Mingmen" promotes the repair of the injured nerve in the spinal anterior horn probably through inhibiting GluR1 expression in the spinal injured area in the rats with acute SCI.

Long non-coding RNA TUG1 knockdown prevents neurons from death to alleviate acute spinal cord injury via the microRNA-338/BIK axis

ABSTRACT Taurine up-regulated gene 1 (TUG1) is a cancer-associated long noncoding RNA (lncRNA) and engages in the development of spinal cord injury (SCI), a suffering neuropathological disorder. However, the regulatory role of TUG1 in acute SCI (ASCI) is still underdetermined. RT-qPCR and western blot analysis were applied to measure the expression of TUG1, microRNA-338 (miR-338), Bcl2-interacting killer (BIK), cleaved caspase 3 (c-caspase 3) and hypoxia-inducible factor-1 alpha (HIF-1α) in ASCI rats and hypoxic cells. Cell death was evaluated using flow cytometric analysis. The relationships among miR-338, TUG1 or BIK were confirmed by luciferase reporter assay, RNA immunoprecipitation and RNA pull-down. Accordingly, we monitored higher expression of TUG1 and BIK, but lower expression of miR-338 in ASCI rats and hypoxic cells. In vitro, hypoxia expedited cell death and c-caspase 3 levels. In vivo, ASCI rats were successfully developed as evidenced by diminished Basso-Beattie-Bresnahan (BBB) locomotor score and enhanced c-caspase 3 and HIF-1α expression. Nevertheless, TUG1 knockdown mitigated the cell death in ASCI rats and hypoxic cells. Mechanically, TUG1 interacted with miR-338 to regulate the BIK expression. Together, TUG1 silencing could alleviate the death in neurons and ASCI models via modulating the miR-338/BIK axis.

Continuous Optical Monitoring of Spinal Cord Oxygenation and Hemodynamics during the First Seven Days Post-Injury in a Porcine Model of Acute Spinal Cord Injury.

One of the only currently available treatment options to potentially improve neurological recovery after acute spinal cord injury (SCI) is augmentation of mean arterial blood pressure (MAP) to promote blood flow and oxygen delivery to the injured cord. However, to optimize such hemodynamic management, clinicians require a method to monitor the physiological effects of these MAP alterations within the injured cord. Therefore, we investigated the feasibility and effectiveness of using a novel optical sensor, based on near-infrared spectroscopy (NIRS), to monitor real-time spinal cord oxygenation and hemodynamics during the first 7 days post-injury in a porcine model of acute SCI. Six Yucatan miniature pigs underwent a T10 vertebral level contusion-compression injury. Spinal cord oxygenation and hemodynamics were continuously monitored by a minimally invasive custom-made NIRS sensor, and by invasive intraparenchymal (IP) probes to validate the NIRS measures. Episodes of MAP alteration and hypoxia were performed acutely after injury, and at 2 and 7 days post-injury to simulate the types of hemodynamic changes SCI patients experience after injury. The NIRS sensor demonstrated the ability to provide oxygenation and hemodynamic measurements over the 7-day post-SCI period. NIRS measures showed statistically significant correlations with each of the invasive IP measures and MAP changes during episodes of MAP alteration and hypoxia throughout the first week post-injury (p < 0.05). These results indicate that this novel NIRS system can monitor real-time changes in spinal cord oxygenation and hemodynamics over the first 7 days post-injury, and has the ability to detect local tissue changes that are reflective of systemic hemodynamic changes.

FTY720 Attenuates Neuropathic Pain after Spinal Cord Injury by Decreasing Systemic and Local Inflammation in a Rat Spinal Cord Compression Model.

Neuropathic pain severely impairs rehabilitation and quality of life after spinal cord injury (SCI). The sphingosine-1-phosphate receptor agonist, FTY720, plays an important protective role in neuronal injury. This study aims to examine the effects of FTY720 in a rat acute SCI model, focusing on neuropathic pain. Female rats with SCI induced by 1-min clip compression were administered vehicle or 1.5 mg/kg of FTY720 24 h after the injury. Using the mechanical nociceptive threshold test, we monitored neuropathic pain and performed histological analysis of the pain pathway, including the μ opioid receptor (MOR), hydroxytryptamine transporter (HTT), and calcitonin gene-related peptide (CGRP). Motor score, SCI lesion volume, residual motor axons, inflammatory response, glial scar, and microvascular endothelial dysfunction were also compared between the two groups. FTY720 treatment resulted in significant attenuation of post-traumatic neuropathic pain. It also decreased systemic and local inflammation, thereby reducing the damaged areas and astrogliosis and resulting in motor functional recovery. Whereas there was no difference in the CGRP expression between the two groups, FTY720 significantly preserved the MOR in both the caudal and rostral areas of the spinal dorsal horn. Whereas HTT was preserved in the FTY720 group, it was significantly increased in the rostral side and decreased in the caudal side of the injury in the vehicle group. These results suggest that FTY720 ameliorates post-traumatic allodynia through regulation of neuroinflammation, maintenance of the blood–brain barrier, and inhibition of glial scar formation, thereby preserving the connectivity of the descending inhibitory pathway and reducing neuropathic pain.

Effects of sacral nerve electrical stimulation on 5‑HT and 5‑HT3AR/5‑HT4R levels in the colon and sacral cord of acute spinal cord injury rat models.

Spinal cord injury (SCI) often leads to defecation dysfunction. Sacral nerve electrical stimulation (SNS) therapy could improve defecation function. The present study aimed to assess SNS therapy, with regard to the levels of serotonin (5-HT) and its receptors (5-HT3AR and 5-HT4R) in the colon and sacral cord, a rat model of acute severe SCI was used. This rat model was made using the New York University Impactor device. Model rats were randomized to the SCI and SNS (electrical stimulation on the S3 nerve) groups. After 14 days of treatment, enteric transmission function was assessed. 5-HT and 5-HT3AR/5-HT4R were measured by ELISA, quantitative PCR, immunohistochemistry and western blotting. In SCI rats, SNS significantly increased the quantity of feces, shortened the time to the first fecal passage, and improved fecal texture and colon histology. SNS elevated 5-HT contents in the colon and spinal cord, and enhanced 5-HT3AR/5-HT4R protein expression and distribution in the colonic myenteric plexus and mucosa, sacral intermediolateral nucleus and dorsal horn. SNS upregulated the relative expression levels of 5-HT3AR/5-HT4R mRNA and protein in the colon and spinal cord. SNS can improve defecation and accelerate the recovery of colonic transmission functions in rat models of acute SCI. These effects involved upregulation of the 5-HT/5-HT3AR/5-HT4R axes.

Expression of CPLX1 in Different Degrees of Spinal Cord Injury in Subacute Stage

This study is to explore the expression of CPLX1 (Complexin 1) in spinal cord tissues with different degrees of injury in the subacute phase of spinal cord injury. Twenty-four SD rats were randomly divided into three experimental groups (light, medium and severe injury) and a sham-operated control group. The experimental group made acute spinal cord injury models according to the lisa method, and the spinal cord tissue were collected 72 hours after injury. The tissues of rats were taken for transcriptome high-throughput sequencing and proteomic analysis experiments to observe the expression of CPLX1 in the different degrees during the subacute phase of spinal cord injury. The mRNA and protein of CPLX1 were detect in rat spinal cord tissue, and they were expressed high in the sham operation group. The protein expression in the light injury group began to decrease, and with the increase of the degree of injury, the transcription level of CPLX1 and the amount of protein expression also gradually decreased. The change trend of mRNA transcription level and protein expression level was basically the same. When the spinal cord injury heavier, the less CPLX1 was expressed, the two showed a significant negative correlation. In the subacute stage of spinal cord injury, the translation and protein levels of CPLX1 decreased significantly. The more severe the injury, the lower the transcription level and protein expression level. CPLX1 may be used as a marker gene for the severity of spinal injury.

Expression of Vimentin in Spinal Cord Injury

This study is to explore the expression differences of vimentin in the normal spinal cord tissues and the injured spinal cord tissues. The 15 SD rats were randomly divided into two experimental groups and sham group. Acute spinal cord injury models in experimental groups were performed with Lisa method and the injured spinal cord was collected after getting injured 72h. The method of immunohistochemical stain was applied to observe the expressions of vimentin in the normal spinal cord tissue and the injured spinal cord tissue of rats. The results showed the expression of vimentin was found in both the grey matter and white matter of the spinal cord of rats with a relatively even distribution. In addition, the protein expressions of vimentin in the medium injury group and the severe injury group were obviously higher than the sham group. In conclusion, expressions of vimentin were found in the spinal cord, and expression differences were observed after the spinal cord injury, which indicated that vimentin might play a part in the injured process of the spinal cord. It is poised to be a phase of advancement in the treatment of acute spinal cord injury and the importance of further elucidation of underlying pathophysiological mechanisms of vimentin.

Efficacy of human HC016 cell transplants on neuroprotection and functional recovery in a rat model of acute spinal cord injury.

Spinal cord injury (SCI) is a devastating event with huge personal and social costs, for which there is no effective treatment. Cell therapy constitutes a promising therapeutic approach for SCI; however, its clinical potential is seriously limited by their low survival in the hostile conditions encompassing the acute phase of SCI. Human HC016 (hHC016) cells, generated from expanded human adipose mesenchymal stem cells (hAMSCs) and pulsed with a patented protocol with hydrogen peroxide (H2 O2 ), are expected to acquire improved resistance to oxidative environments which appears as a major limiting factor hampering the engrafting success. Our specific aim was to assess whether H2 O2 -pulsed hHC016 cells had an improved survival and thus therapeutic efficacy in a rat contusion model of acute SCI when grafted 48 hr after injury. Functional recovery was evaluated up to 56 days post-injury (dpi) by locomotor (open field test and CatWalk) and sensory (Von Frey and Hargreaves) tests. Besides, histological evaluation of transplanted cell survival and tissue protection/regeneration was also performed. Functional results showed a statistically significant improvement on locomotor recovery outcomes with hHC016 cells. Accordingly, superior cell survival in correlation with long-term neuroprotection, higher axonal regeneration, and reduced astroglial and microglial reactivity was also observed with hHC016 cells. These results demonstrate an enhanced survival capacity of hHC016 cells resulting in improved functional and histological outcomes as compared with hAMSCs, indicating that hHC016 cell transplants may constitute a promising cell therapy for acute SCI.

Melatonin exerts neuroprotective effects by attenuating astro- and microgliosis and suppressing inflammatory response following spinal cord injury

Reactive gliosis and inflammatory reaction are common pathological change to spinal cord injury (SCI). Whereas, the effects of melatonin (MT) on the astro- and microgliosis and their related inflammatory response in the injured spinal cord are not fully understood. In this study, MT's effects on the accumulation and proliferation of microglia and astrocytes and their related inflammatory response were investigated in an acute SCI model. The effects of MT on oxidative stress, neuronal survival and behavioral performance were also tested. It was found that MT treatment significantly suppressed the accumulation and the proliferation of microglia and astrocytes. Quantitative PCR data showed that MT significantly down-regulated the pro-inflammatory markers iNOS, IL-1β and TNF-α expressions. The data showed that MT led to the rise in SOD, CAT and GSH-Px contents and the decrease in MDA content. Western blotting analysis verified that MT significantly down-regulated caspase-3, Bax and GFAP expressions, up-regulated Bcl-2 expression. Compared with the SCI vehicle-treated group, the SCI MT-treated group exhibited a greater Basso Mouse Scale (BMS) locomotor rating score. On the whole, these findings implied that MT exerts its neuroprotective effects by suppressing the accumulation and the proliferation of microglia and astrocytes and reducing the release of pro-inflammatory cytokines, which might be one of the underlying mechanisms of the MT's neuroprotective effect after SCI. Accordingly, MT may be a promising therapeutic candidate for acute SCI.

Optical Assessment of Spinal Cord Tissue Oxygenation Using a Miniaturized Near Infrared Spectroscopy Sensor.

Despite advances in the treatment of acute spinal cord injury (SCI), measures to mitigate permanent neurological deficits in affected patients are limited. Immediate post-trauma hemodynamic management of patients, to maintain blood supply and improve oxygenation to the injured spinal cord, is currently one aspect of critical care which clinicians can utilize to improve neurological outcomes. However, without a way to monitor the response of spinal cord hemodynamics and oxygenation in real time, optimizing hemodynamic management is challenging and limited in scope. This study aims to investigate the feasibility and validity of using a miniaturized multi-wavelength near-infrared spectroscopy (NIRS) sensor for direct transdural monitoring of spinal cord oxygenation in an animal model of acute SCI. Nine Yorkshire pigs underwent a weight-drop T10 contusion-compression injury and received episodes of ventilatory hypoxia and alterations in mean arterial pressure (MAP). Spinal cord hemodynamics and oxygenation were monitored throughout by a non-invasive transdural NIRS sensor, as well as an invasive intraparenchymal sensor as a comparison. NIRS parameters of tissue oxygenation were highly correlated with intraparenchymal measures of tissue oxygenation. In particular, during periods of hypoxia and MAP alterations, changes of NIRS-derived spinal cord oxygenated hemoglobin and tissue oxygenation percentage corresponded well with the changes in spinal cord oxygen partial pressures measured by the intraparenchymal sensor. Our data confirm that during hypoxic episodes and as changes occur in the MAP, non-invasive NIRS can detect and measure real-time changes in spinal cord oxygenation with a high degree of sensitivity and specificity.