Model Of Traumatic Spinal Cord Injury Research Articles

Effect of humic acid on oxidative stress and neuroprotection in traumatic spinal cord injury: an experimental study.

Traumatic spinal cord injury (TSCI) is an important health problem, especially in developing countries with additional socioeconomic loss. Humic acid (HA) usually has antioxidant, antiinflammatory, blood circulating, and antiviral effects. Hence, it was aimed herein to show the effect of HA on neuroprotection in a TSCI model. A TSCI model was used, in which 24 Wistar albino rats were divided into 4 groups: control group: subjected to only laminectomy; sham group: subjected to laminectomy + TSCI; HA 5 mg/kg group: subjected to laminectomy + TSCI + intraperitoneal (IP) injection of 5 mg/kg of HA; and HA 10 mg/kg group: subjected to laminectomy + TSCI + IP injection of 10 mg/kg of HA. Intracardiac blood samples were obtained preoperatively (preop), and at 1 and 24 h postoperatively (postop). The total antioxidant status (TAS), total oxidant status (TOS) and oxidative stress index (OSI) levels were evaluated in the serum. The motor functions were evaluated using the Modified Tarlov Score at 24 h postop. There were no significant changes in the TAS values between the sham and HA 5 mg/kg and HA 10 mg/kg groups (p = 0.77/0.21). However there was a significant decrease in the TOS values at 24 h postop when comparing the sham and HA 5 mg/kg groups (p = 0.02). The pathological evaluation showed a significant decrease in the severity of edema, hemorrhage, polymorphonuclear leucocyte (PNL) infiltration, and mononuclear leucocyte (MNL)/macrophage/microglia infiltration when compared with the control group (p < 0.05). There was a significant recovery at the paraplegia level when the HA 5 mg/kg and HA 10 mg/kg groups were compared with the control group (p < 0.001). The effects of HA in the early stages of TSCI on oxidative stress, histopathological changes, and neurological improvement were investigated herein. It is thought to be a potential therapeutic agent in acute TSCI but needs to be further evaluated to determine the extent of its effect on other neuroprotective pathways in larger series.

A 72-h sedated porcine model of traumatic spinal cord injury

IntroductionThere is an increasing focus on the prevention of secondary injuries following traumatic spinal cord injury (TSCI), especially through improvement of spinal cord perfusion and immunological modulation. Such therapeutic strategies require translational and controlled animal models of disease progression of the acute phases of human TSCI. Research questionIs it possible to establish a 72-h sedated porcine model of incomplete thoracic TSCI, enabling controlled use of continuous, invasive, and non-invasive modalities during the entire sub-acute phase of TSCI? Material and methodsA sham-controlled trial was conducted to establish the model, and 10 animals were assigned to either sham or TSCI. All animals underwent a laminectomy, and animals in the TSCI group were subjected to a weight-drop injury. Animals were then kept sedated for 72 h. The amount of injury was assessed by ex-vivo measures MRI-based fiber tractography, histology and immunohistochemistry. ResultsIn all animals, we were successful in maintaining sedation for 72 h without comprising vital physiological parameters. The MRI-based fiber tractography showed that all TSCI animals revealed a break in the integrity of spinal neurons, whereas histology demonstrated no transversal sections of the spine with complete injury. Notably, some animals displayed signs of secondary ischemic tissue in the cranial and caudal sections. Discussion and conclusionsThis study succeeded in producing a porcine model of incomplete TSCI, which was physiologically stable up to 72 h. We believe that this TSCI model will constitute a potential translational model to study the pathophysiology secondary to TSCI in humans.

HSPA1A ameliorated spinal cord injury in rats by inhibiting apoptosis to exert neuroprotective effects.

Traumatic spinal cord injury (TSCI) is a serious nervous system insult, and apoptosis in secondary injury is an important barrier to recovery from TSCI. Heat shock protein family A member 1A (HSPA1A) is a protective protein whose expression is elevated after stress. However, whether HSPA1A can inhibit apoptosis after spinal cord injury, and the potential mechanism of this inhibition, remain unclear. In this study, we established in vivo and in vitro models of TSCI and induced HSPA1A overexpression and silencing. HSPA1A upregulation promoted the recovery of neurological function and pathological morphology at the injury site, enhanced neurological cell survival, and inhibited apoptosis in rats following TSCI. In the in vitro model, HSPA1A overexpression inhibited H2O2-induced apoptosis, indicating that HSPA1A suppressed the expression of Bax, caspase-9, and cleaved-caspase-3, promoted the expression of Bcl-2. Furthermore, inhibition of HSPA1A expression can aggravate H2O2-induced apoptosis. We also found that HSPA1A overexpression activated the Wnt/β-catenin signaling pathway, and that inhibition of this pathway attenuated the inhibitory effect of HSPA1A overexpression on apoptosis. Together, these results indicate that HSPA1A has neuroprotective effects against TSCI that may be exerted through activation of the Wnt/β-catenin signaling pathway to inhibit apoptosis.

Hydrogen Sulfide Improves Functional Recovery in Rat Traumatic Spinal Cord Injury Model by Inducing Nuclear Translocation of NF-E2-Related Factor 2.

Hydrogen sulfide (H2S), an important gaseous messenger, is known to have neuroprotective effects in many neurological disorders. This study examined the neuroprotective effects and the associated mechanisms of H2S in the model Sprague-Dawley (SD) rats with spinal cord injury (SCI). We found that H2S showed neuroprotective effects in SCI model rats, improved the symptoms of neurological impairment, reduced the secretion of inflammatory factors, nerve cell apoptosis, and endoplasmic reticulum (ER), and oxidative stresses. Moreover, these effects were produced by activation of nuclear factor-erythroid 2-related factor 2 (Nrf2) protein. Our results suggest that H2S supplementation could be a potential therapeutic strategy to promote SCI recovery.

Effect of Humic Acid on Oxidative Stress and Neuroprotection in Traumatic Spinal Cord Injury: An Experimental Study.

Traumatic spinal cord injury (TSCI) is an important health problem especially in developing countries with additional socio-economic loss. Humic acids (HA) usually have anti-oxidant, anti-inflammatory, blood-circulating and antiviral effects. We aimed to show effect of HA on neuroprotection in TSCI model. We performed TSCI model in Twenty-four Wistar-Albino rats in four groups. Control group underwent only laminectomy. Sham group underwent laminectomy followed by TSCI. Low dose HA (5mg/kg) and high dose HA (10mg/kg) groups underwent laminectomy and TSCI followed by peritoneal administration of HA. Preoperative, postoperative 1st hour and postoperative 24th hour cardiac blood samples were obtained. Total Antioxidant Status (TAS), Total Oxidant Status (TOS) and Oxidative Index (OI) levels were evaluated in serum. The 24th hour motor functions were evaluated by Modified Tarlov Score. There were no significant changes in TAS values between sham- low dose and high dose humic acid groups (p:0.77/0.21). However there were a significant decrease of TOS levels in the 24th hour post operative blood samples comparing the sham group with low dose humic acid group (p=0.02). Pathological evaluation showed a significant decrease in the severity of edema, hemorrhage, Polymorphonuclear leucocytes (PNL) and Mononuclearleucocytes (MNL) /macrophage/microglia when we compare with the control group (p<0.05). There is a significant recovery in paraplegia level as we compared the HA groups with control groups (p<0.001). In this study, we showed the effects of HA in the early stages of TSCI on oxidative stress, histopathological changes and neurological improvement. It is thought to be a potential therapeutic agent in acute TSCI but needs to be further evaluated by showing proper effect on other neuroprotective pathways in larger series.

Role of circular RNA expression in the pathological progression after spinal cord injury.

Differential expression of non-coding RNA after traumatic spinal cord injury (TSCI) is closely related to the pathophysiological process. The purposes of this study were to systematically profile and characterize expression of circular RNA (circRNA) in the lesion epicenter of spinal tissues after TSCI, and predict the structure and potential function of the regulatory circRNA/miRNA network. Forty-eight C57BL/6 mice were randomly and equally assigned to two groups: one subjected to TSCI at T8-10 with an Allen's drop impactor, and a second subjected to laminectomy without TSCI. Spinal cord samples were stained with hematoxylin and eosin, sequenced, and validated. RNA-Seq, Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes analysis, and network analyses (Targetscan and miRanda) were used to predict and annotate the circRNA/miRNA/mRNA network. Luciferase reporter, quantitative reverse transcription polymerase chain reaction, and western blot assays were used to profile expression and regulation patterns of the network in mouse models of TSCI. Hematoxylin-eosin staining revealed severe damage to the blood-spinal cord barrier after TSCI. Differentially expressed circRNA and miRNA profiles were obtained after TSCI; differentially expressed circRNAs, which were abundant in the cytoplasm, were involved in positive regulation of transcription and protein phosphorylation. miR-135b-5p was the most significantly downregulated miRNA after TSCI; circRNAAbca1 and KLF4 were predicted to be its target circRNA and mRNA, respectively. Subsequently, the circAbca1/miR-135b-5P/KLF4 regulatory axis was predicted and constructed, and its targeted binding was verified. After inhibiting circAbca1, GAP43 expression was upregulated. Differential expression of circRNAs might play an important role after TSCI. circAbca1 plays a neuroinhibitory role by targeted binding of the miR-135b-5P/KLF4 axis. The identified circRNA/miRNA/mRNA network could provide the basis for understanding pathophysiological mechanisms underlying TSCI, as well as guide the formulation of related therapeutic strategies. All animal protocols were approved by the Research Ethics Committee of West China Hospital of China (approval No. 2017128) on May 16, 2017.

Inhibition of leucine-rich repeats and calponin homology domain containing 1 accelerates microglia-mediated neuroinflammation in a rat traumatic spinal cord injury model

BackgroundSpinal cord injury (SCI) triggers the primary mechanical injury and secondary inflammation-mediated injury. Neuroinflammation-mediated insult causes secondary and extensive neurological damage after SCI. Microglia play a pivotal role in the initiation and progression of post-SCI neuroinflammation.MethodsTo elucidate the significance of LRCH1 to microglial functions, we applied lentivirus-induced LRCH1 knockdown in primary microglia culture and tested the role of LRCH1 in microglia-mediated inflammatory reaction both in vitro and in a rat SCI model.ResultsWe found that LRCH1 was downregulated in microglia after traumatic SCI. LRCH1 knockdown increased the production of pro-inflammatory cytokines such as IL-1β, TNF-α, and IL-6 after in vitro priming with lipopolysaccharide and adenosine triphosphate. Furthermore, LRCH1 knockdown promoted the priming-induced microglial polarization towards the pro-inflammatory inducible nitric oxide synthase (iNOS)-expressing microglia. LRCH1 knockdown also enhanced microglia-mediated N27 neuron death after priming. Further analysis revealed that LRCH1 knockdown increased priming-induced activation of p38 mitogen-activated protein kinase (MAPK) and Erk1/2 signaling, which are crucial to the inflammatory response of microglia. When LRCH1-knockdown microglia were adoptively injected into rat spinal cords, they enhanced post-SCI production of pro-inflammatory cytokines, increased SCI-induced recruitment of leukocytes, aggravated SCI-induced tissue damage and neuronal death, and worsened the locomotor function.ConclusionOur study reveals for the first time that LRCH1 serves as a negative regulator of microglia-mediated neuroinflammation after SCI and provides clues for developing novel therapeutic approaches against SCI.

Characterization of traumatic spinal cord injury model in relation to neuropathic pain in the rat

Purpose/aim: Neuropathic pain following spinal cord injury (SCI) has a tremendous impact on patient’s quality of life, and frequently is the most limiting aspect of the disease. In view of the severity of this condition and the absence of effective treatments, the establishment of a reliable animal model that reproduces neuropathic pain after injury is crucial for a better understanding of the pathophysiology and for the development of new therapeutic strategies. Thus, the objective of the present study was to standardize the traumatic SCI model in relation to neuropathic pain.Materials and methods: Wistar rats were submitted to SCI of mild intensity (pendulum height 12.5 mm) or moderate intensity (pendulum height 25 mm) using the New York University Impactor equipment. Behavioural assessment was performed during 8 weeks. Thereafter, spinal cords were processed for immunohistochemistry.Results: The animals of the moderate injury group in comparison with mild injury had a greater motor function deficit, worse mechanical allodynia, and latter bladder recovery; moreover, histological analysis revealed more extensive lesions with lower neuronal population.Conclusions: Our study suggests that moderate SCI causes a progressive and long-lasting painful condition (at least 8 weeks), in addition to motor impairment, and thus represents a reliable animal model for the study of chronic neuropathic pain after SCI.

Allicin protects traumatic spinal cord injury through regulating the HSP70/Akt/iNOS pathway in mice.

Allicin is a major component of garlic, extracted as an oily liquid. The present study was designed to investigate the beneficial effects of allicin on traumatic spinal cord injury (TSCI) in mice, and whether the effects are mediated via regulation of the heat shock protein 70 (HSP70), v-akt murine thymoma viral oncogene homolog 1 (Akt) and inducible nitric oxide synthase (iNOS) pathways. Adult BALB/c mice (30–40 g) received a laminectomy at the T9 vertebral level as a model of TSCI. In the present study, treatment of the TSCI mice with allicin significantly increased their Basso, Beattie and Bresnahan (BBB) scores (P<0.01) and reduced the spinal cord water content (P<0.01). This protective effect was associated with the inhibition of oxidative stress and inflammatory responses in TSCI mice. Western blot analysis demonstrated that allicin increased the protein levels of HSP70, increased the phosphorylation of Akt and reduced the iNOS protein expression levels in TSCI mice. Additionally, treatment with allicin significantly reduced the levels of ROS and enhanced the NADH levels in TSCI mice. Collectively, these data demonstrate that the effects of allicin on TSCI are mediated via regulation of the HSP70, Akt and iNOS pathways in mice.

Lipopolysaccharide–induced preconditioning protects against traumatic spinal cord injury by upregulating Nrf2 expression in rats

AimsLipopolysaccharide (LPS)-induced preconditioning protects neurons against traumatic spinal cord injury (TSCI) in rats. This study sought to test whether Nrf2, a transcription factor, mediated LPS-induced preconditioning. Main methodsThe TSCI model was established using a standardized NYU impactor on adult female rats. Rats were pretreated with LPS (0.2mg/kg, IP; 72h before injury). Nrf2 was silenced by injecting a lentivirus encoding RNAi against Nrf2 into injured spinal cords. Neurologic function was assessed by Basso, Beattie and Bresnahan (BBB) scores 6h, 12h, 24h, 48h, 72h, 7d and 14d after TSCI. Neuronal apoptosis was measured by a TUNEL staining. Ultrastructure was observed using transmission electron microscope (TEM). The protein expression of HO-1, NQO1 and GCLC was examined using immunohistochemistry and immunoblotting. Key findingsThe injection of a lentivirus effectively transfected GFP into injured spinal cords. The expression of Nrf2 was significantly decreased in spinal cords receiving a lentivirus encoding RNAi against Nrf2. BBB scores showed that TSCI markedly impaired nervous function, which was markedly preserved by LPS pretreatment. Nrf2 knockdown significantly suppressed LPS pretreatment–induced protection of nervous function. TEM images and TUNEL staining showed an increase in apoptotic cells when Nrf2 was silenced. Moreover, immunohistochemistry and immunoblotting analysis exhibited that LPS pretreatment significantly upregulated the expression of anti–oxidative proteins including HO-1, NQO1 and GCLC, which was suppressed when Nrf2 expression was silenced in injured spinal cords. SignificanceLPS preconditioning effectively alleviates TSCI–induced impairment and preserves nervous function in a Nrf2–dependent manner.

Estimating the global incidence of traumatic spinal cord injury

Population modelling--forecasting. To estimate the global incidence of traumatic spinal cord injury (TSCI). An initiative of the International Spinal Cord Society (ISCoS) Prevention Committee. Regression techniques were used to derive regional and global estimates of TSCI incidence. Using the findings of 31 published studies, a regression model was fitted using a known number of TSCI cases as the dependent variable and the population at risk as the single independent variable. In the process of deriving TSCI incidence, an alternative TSCI model was specified in an attempt to arrive at an optimal way of estimating the global incidence of TSCI. The global incidence of TSCI was estimated to be 23 cases per 1,000,000 persons in 2007 (179,312 cases per annum). World Health Organization's regional results are provided. Understanding the incidence of TSCI is important for health service planning and for the determination of injury prevention priorities. In the absence of high-quality epidemiological studies of TSCI in each country, the estimation of TSCI obtained through population modelling can be used to overcome known deficits in global spinal cord injury (SCI) data. The incidence of TSCI is context specific, and an alternative regression model demonstrated how TSCI incidence estimates could be improved with additional data. The results highlight the need for data standardisation and comprehensive reporting of national level TSCI data. A step-wise approach from the collation of conventional epidemiological data through to population modelling is suggested.

The place escape/avoidance paradigm: A novel method to assess nociceptive processing

This paper summarizes a behavioral paradigm that was developed as a novel method to dissociate the multidimensional pain experience in rodents. The place escape/avoidance paradigm (PEAP) is based on the assumption that if animals escape and/or avoid a noxious stimulus, then the stimulus is aversive to the animal. Data is presented showing that when animals are placed in a specific environmental condition, they will perform purposeful behavior to escape and/or avoid the noxious stimulus. Additional data is presented to demonstrate the validity of the behavioral paradigm and how the paradigm has been used to test the hypothesis that the affective/motivational dimension of pain can be dissociated and studied independent of sensory pain processing. The behavioral paradigm highlights the emerging trend in the area of pain research and management towards developing more realistic behavioral paradigms to assess nociceptive processing in rodent models of chronic pain.

Lentivirus-siNgR199 Promotes Axonal Regeneration and Functional Recovery in Rats

ABSTRACTBackground: The limited regenerative capability of central neurons and inhibitory factors are the main causes of axonal regeneration failure after spinal cord injury (SCI). Nogo receptors (NgR) are a family of receptors shared by three factors that inhibit axon outgrowth. In a previous study, siNgR199, an effective lentiviral siRNA vector of NgR1, was constructed and transfected into cortical neurons in vitro, and it effectively promoted axon outgrowth. The present study focused on the therapeutic effect of delivery of a recombinant lentivirus containing siNgR199 in vivo in rats. Methods: Rat models of traumatic SCI were constructed according to the method developed by Allen. The animals were randomly divided into three groups: group 1, injected with physiological saline; group 2, injected with empty lentivirus vehicle; and group 3, injected with lentivirus carrying siNgR199. The Basso, Beattie, and Bresnahan (BBB) scale was used for assessing hindlimb locomotor function after SCI. The neural tracer biotinylated dextran amine (BDA) and immunohistochemical methods were employed to study axon outgrowth. Results: After injection for 8 weeks, BBB locomotion scores showed that the motor function of the hindlimb recovered better in animals in group 3 (injected with lentivirus carrying siNgR199) than those in groups 1 and 2, which were injected with saline and empty lentivirus vehicle, respectively. In group 3, regenerative nerve fibers were observed at and across the injury site, while very few axonal sprouts were observed in groups 1 and 2. Conclusions: Injection with lentivirus carrying siNgR199 into the sensorimotor cortex improved axonal regeneration and functional recovery of hindlimb after SCI in rats.

Glial scar and neuroregeneration: histological, functional, and magnetic resonance imaging analysis in chronic spinal cord injury

A glial scar is thought to be responsible for halting neuroregeneration following spinal cord injury (SCI). However, little quantitative evidence has been provided to show the relationship of a glial scar and axonal regrowth after injury. In this study performed in rats and dogs, a traumatic SCI model was made using a weight-drop injury device, and tissue sections were stained with H & E for immunohistochemical analysis. The function and behavior of model animals were tested using electrophysiological recording and the Basso-Beattie-Bresnahan Locomotor Rating Scale, respectively. The cavity in the spinal cord after SCI in dogs was observed using MR imaging. The morphological results showed that the formation of an astroglial scar was defined at 4 weeks after SCI. While regenerative axons reached the vicinity of the lesion site, the glial scar blocked the extension of regrown axons. In agreement with these findings, the electrophysiological, behavioral, and in vivo MR imaging tests showed that functional recovery reached a plateau at 4 weeks after SCI. The thickness of the glial scars in the injured rat spinal cords was also measured. The mean thickness of the glial scar rostral and caudal to the lesion cavity was 107.00 +/- 20.12 microm; laterally it was 69.92 +/- 15.12 microm. These results provide comprehensive evidence indicating that the formation of a glial scar inhibits axonal regeneration at 4 weeks after SCI. This study reveals a critical time window of postinjury recovery and a detailed spatial orientation of glial scar, which would provide an important basis for the development of therapeutic strategy for glial scar ablation.

Reduction of hypoxia by perfluorocarbon emulsion in a traumatic spinal cord injury model

Few therapies have consistently demonstrated effectiveness in preserving O2 delivery after spinal cord injury (SCI). Perfluorocarbons (PFCs) offer great promise to carry and deliver O2 more efficiently than conventional measures. The authors investigated the use of Clark-type microelectrodes to monitor spinal cord oxygenation directly (intraparenchymal [IP] recording) and indirectly (cerebrospinal fluid [CSF] recording) in the context of SCI, O2 therapy, and PFC treatment. After placement of a subdural/CSF Licox probe in rats, incremental increases in the fraction of inspired O2 (FiO2) up to 100% were administered to establish a dose-response curve. The probe was then placed in the parenchyma of the same animals for a second dose-response curve. In a second study, rats with CSF or IP probes underwent SCI with the NYU Impactor and treatment with O2, followed by administration of PFC, or saline in the control group. All animals in the first experiment responded to the FiO2 dose increase, with changes in PO2 evident in both CSF and IP levels. The SCI in the second experiment caused a marked drop in PO2 from a mean of 21.4 to 10.4 mm Hg, with most animals dropping to less than half their preinjury value. All animals responded to 100% O2 treatment. Every animal that received PFCs showed significant improvement, with a mean increase in PO2 of 23.3 mm Hg. Only 1 saline-treated animal showed any benefit. Oxygen values in the PFC treatment group reached up to 6 times the normal level. Oxygen levels in SCI show a profound drop almost immediately postinjury. Administration of PFCs combined with 100% O2 therapy can reverse tissue hypoxia and holds promise for reducing ischemic injury.

Proliferation of NG2-Positive Cells and Altered Oligodendrocyte Numbers in the Contused Rat Spinal Cord

Given the numerous reparative roles glia may play after spinal cord injury (SCI), glial proliferation and cell number were examined in a model of traumatic SCI. Emphasis was placed on analysis of oligodendrocytes and NG2-positive (NG2+) cells, an endogenous cell population that may be involved in oligodendrocyte replacement. Overall, proliferation (assessed by bromodeoxyuridine incorporation) was markedly elevated during the first 2 weeks after injury and declined thereafter; a large portion of these dividing cells likely consisted of microglia-macrophages. Although the total number of NG2+ cells in the epicenter was reduced by half, we noted protracted proliferation in surviving NG2+ cells, with values sevenfold greater than in uninjured controls. Elevated proliferation of NG2+ cells persisted throughout the first 4 weeks after injury. However, the absolute number of NG2+ cells was not increased over controls, suggesting that the daughter cells either did not survive or they differentiated into other cell types. As expected, oligodendrocyte numbers were drastically altered after SCI. By 7 d after injury, the number of oligodendrocytes at the impact site was reduced by 93%. Despite ongoing tissue loss, the number of oligodendrocytes in spared tissue rose threefold at 14 d after injury. Although the function of NG2+ cells within the spinal cord is not completely understood, several studies suggest that they may differentiate into oligodendrocytes. Thus, proliferating NG2+ cells may contribute to the increased oligodendrocyte number observed at 2 weeks after injury. Future studies are required, however, to definitively determine the role NG2+ cells play in oligodendrocyte genesis, remyelination, and other post-injury events.