The secondary injury cascade after spinal cord injury:an analysis of local cytokine/chemokine regulation

After spinal cord injury,there is an extensive infiltration of immune cells,which exacerbates the injury and leads to further neural degeneration.Therefore,a major aim of current research involves targeting the immune response as a treatment for spinal cord injury.Although much research has been performed analyzing the complex inflammatory process following spinal cord injury,there remain major discrepancies within previous literature regarding the timeline of local cytokine regulation.The objectives of this study were to establish an overview of the timeline of cytokine regulation for 2 weeks after spinal cord injury,identify sexual dimorphisms in terms of cytokine levels,and determine local cytokines that significantly change based on the severity of spinal cord injury.Rats were inflicted with either a mild contusion,moderate contusion,severe contusion,or complete transection,7 mm of spinal cord centered on the injury was harvested at varying times post-injury,and tissue homogenates were analyzed with a Cytokine/Chemokine 27-Plex assay.Results demonstrated pro-inflammatory cytokines including tumor necrosis factorα,interleukin-1β,and interleukin-6 were all upregulated after spinal cord injury,but returned to uninjured levels within approximately 24 hours post-injury,while chemokines including monocyte chemoattractant protein-1 remained upregulated for days post-injury.In contrast,several anti-inflammatory cytokines and growth factors including interleukin-10 and vascular endothelial growth factor were downregulated by 7 days post-injury.After spinal cord injury,tissue inhibitor of metalloproteinase-1,which specifically affects astrocytes involved in glial scar development,increased more than all other cytokines tested,reaching 26.9-fold higher than uninjured rats.After a mild injury,11 cytokines demonstrated sexual dimorphisms;however,after a severe contusion only leptin levels were different between female and male rats.In conclusion,pro-inflammatory cytokines initiate the inflammatory process and return to baseline within hours post-injury,chemokines continue to recruit immune cells for days post-injury,while anti-inflammatory cytokines are downregulated by a week post-injury,and sexual dimorphisms observed after mild injury subsided with more severe injuries.Results from this work define critical chemokines that influence immune cell infiltration and important cytokines involved in glial scar development after spinal cord injury,which are essential for researchers developing treatments targeting secondary damage after spinal cord injury.

DTI and pathological changes in a rabbit model of radiation injury to the spinal cord after ^(125)I radioactive seed implantation

Excessive radiation exposure may lead to edema of the spinal cord and deterioration of the nervous system. Magnetic resonance imaging can be used to judge and assess the extent of edema and to evaluate pathological changes and thus may be used for the evaluation of spinal cord injuries caused by radiation therapy. Radioactive ^125I seeds to irradiate 90% of the spinal cord tissue at doses of 40–100 Gy （D90） were implanted in rabbits at T10 to induce radiation injury, and we evaluated their safety for use in the spinal cord. Diffusion tensor imaging showed that with increased D90, the apparent diffusion coefficient and fractional anisotropy values were increased. Moreover, pathological damage of neurons and microvessels in the gray matter and white matter was aggravated. At 2 months after implantation, obvious pathological injury was visible in the spinal cords of each group. Magnetic resonance diffusion tensor imaging revealed the radiation injury to the spinal cord, and we quantified the degree of spinal cord injury through apparent diffusion coefficient and fractional anisotropy.

Local low-dose X-ray radiation promotes homing of mesenchymal stem cells to the injured mouse spinal cord

BACKGROUND： Bone marrow-derived mesenchymal stem cells （BMSCs） are a potentially useful source for cell replacement therapy following spinal cord injury. However, the homing characteristics of BMSCs in vivo remain unclear. Low-dose radiation has been shown to promote homing of BMSCs to exposed sites. OBJECTIVE： To investigate the effects of low-dose local radiation to non-injured areas on the ability of human BMSCs to home to the injured mouse spinal cord, as well as recovery of spinal cord injury. DESIGN, TIME AND SE＇I-FING： A randomized, controlled, animal experiment was performed at the Central Laboratory, Second Affiliated Hospital of Soochow University between October 2007 and October 2008. MATERIALS： BMSCs were isolated from four adult, human donors. METHODS： Fifty adult, female, Balb/c mice were subjected to adjusted weight-drop impact resulting in complete paraplegia. Three days later, mice were randomly assigned to a radiation ＋ transplantation group （n = 23） and a transplantation group （n = 20）. In total, 2 x 106 carboxyfluorescein diacetate succinimidyl ester-labeled BMSCs were injected into each mouse via the caudal vein. Mice in the radiation ＋ transplantation group received 2.5 Gy local X-ray irradiation 2 hours before BMSCs injection. MAIN OUTCOME MEASURES： The homing of BMSCs to injured cord and irradiated skin after transplantation was observed by fluorescence microscope; the structure recovery of injured cord was assessed by magnetic resonance imaging. RESULTS： Compared with the transplantation group, at 24 hours after transplantation, the number of BMSCs was significantly increased in the injured area and the exposed site （P 〈 0.05）, and inflammation and edema were significantly alleviated in the injured cord in the radiation ＋ transplantation group. CONCLUSION： Local low-dose radiation has the potential to promote homing of BMSCs and recovery of spinal cord injury, although the radiated region was not injured area.

Reduction of epinephrine in the lumbar spinal cord following repetitive blast-induced traumatic brain injury in rats

Traumatic brain inju ry-induced unfavorable outcomes in human patients have independently been associated with dysregulated levels of monoamines,especially epinephrine,although few preclinical studies have examined the epinephrine level in the central nervous system after traumatic brain injury.Epinephrine has been shown to regulate the activities of spinal motoneurons as well as increase the heart rate,blood pressure,and blood flow to the hindlimb muscles.Therefore,the purpose of the present study was to determine the impact of repeated blast-induced traumatic brain injury on the epinephrine levels in seve ral function-s pecific central nervous system regions in rats.Following three repeated blast injuries at 3-day intervals,the hippocampus,motor cortex,locus coeruleus,vestibular nuclei,and lumbar spinal cord were harvested at post-injury day eight and processed for epinephrine assays using a high-sensitive electrochemical detector cou pled with high-performance liquid chromatography.Our results showed that the epinephrine levels were significantly decreased in the lumbar spinal cord tissues of blast-induced traumatic brain injury animals compared to the levels detected in age-and sex-matched sham controls.In other function-specific central nervous system regions,although the epinephrine levels were slightly altered following blast-induced tra u matic brain injury,they were not statistically significant.These results suggest that blast injury-induced significant downregulation of epinephrine in the lumbar spinal cord could negatively impact the motor and cardiovascular function.This is the first repo rt to show altered epinephrine levels in the spinal cord following repetitive mild blast-induced traumatic brain injury.

Spinal cord biological safety of image-guided radiation therapy versus conventional radiation therapy

Tumor models were simulated in purebred Beagles at the T9-10 levels of the spinal cord and treated with spinal image-guided radiation therapy or conventional radiation therapy with 50 or 70 Gy total radiation. Three months after radiation, neuronal injury at the T9-10 levels was observed, including reversible injury induced by spinal image-guided radiation therapy and apoptosis induced by conventional radiation therapy. The number of apoptotic cells and expression of the proapoptotic protein Fas were significantly reduced, but expression of the anti-apoptotic protein heat shock protein 70 was significantly increased after image-guided radiation therapy compared with the conventional method of the same radiation dose. Moreover, the spinal cord cell apoptotic index positively correlated with the ratio of Fas/heat shock protein 70. These findings indicate that 3 months of radiation therapy can induce a late response in the spinal cord to radiation therapy; image-guided radiation therapy is safer and results in less neuronal injury compared with conventional radiation therapy.

Melatonin for the treatment of spinal cord injury

Spinal cord injury（SCI） from trauma or disease severely impairs sensory and motor function. Neurorehabilitation after SCI is a complex medical process that focuses on improving neurologic function and repairing damaged connections in the central nervous system. An increasing number of preclinical studies suggest that melatonin may be useful for the treatment of SCI. Melatonin is an indolamine that is primarily secreted by the pineal gland and known to be regulated by photoperiodicity. However, it is also a versatile hormone with antioxidative, antiapoptotic, neuroprotective, and anti-inflammatory properties. Here, we review the neuroprotective properties of melatonin and the potential mechanisms by which it might be beneficial in the treatment of SCI. We also describe therapies that combine melatonin with exercise, oxytetracycline, and dexamethasone to attenuate the secondary injury after SCI and limit potential side effects. Finally, we discuss how injury at different spinal levels may differentially affect the secretion of melatonin.

Low-dose radiation on spinal cord might be a new therapy for intractable chronic cancer and non-cancer pain

Intractable chronic pain is a great challenge in clinic.Central sensitization based on the positive changes of dendritic spines is the main mechanism of intractable chronic pain.And low-dose radiation has been proved to regulate the changes of dendritic spines negatively.Hence,we make a hypothesis that low-dose radiation could relieve cancer and noncancer pain through negatively regulating the shape and reducing the number and density of dendritic spines in the spinal cord.This method is supposed to be a new therapy for intractable chronic pain by expanding indication to non-cancer pain,translocating radiation site from where the tumor exists to special segments of spinal cord and keeping radiation dose at a low level.This therapy would be reliable for relieving non-cancer pain and supply more choices for relieving cancer pain.

Radiation-Induced Spinal Glioblastoma Multiforme: A Rare Complication in the Management of Head and Neck Cancer

Background: Radiation-induced gliomas of the spinal cord are rare late complications of spinal cord irradiation that typically occur in patients treated at younger ages. Aim: Raise awareness of radiation induced high grade gliomas with a case presentation and a review of the literature. Case Presentation: A 50-year-old male with Stage IVA squamous cell carcinoma of the oropharynx was treated with external beam radiotherapy with a complete response. Seven years later, he presented with a cervical spinal cord mass on MRI. An open biopsy was performed. Pathology revealed an intramedullary WHO grade IV astrocytoma, (i.e., glioblastoma multiforme) of the cervical spine that fulfilled the criteria for a radiation-induced malignancy. Conclusions : Review of the literature suggests that radiation-induced gliomas tend to be high grade and may occur at the periphery of an irradiated field. Radiation-induced gliomas of the spinal cord are a serious complication of radiotherapy that may occur in older patients with head and neck cancers, but are so rare that it should not affect treatment decisions.

Function of microglia and macrophages in secondary damage after spinal cord injury

Spinal cord injury （SCI） is a devastating type of neurological trauma with limited therapeutic op- portunities. The pathophysiology of SCI involves primary and secondary mechanisms of injury. Among all the secondary injury mechanisms, the inflammatory response is the major contrib- utor and results in expansion of the lesion and further loss of neurologic function. Meanwhile, the inflammation directly and indirectly dominates the outcomes of SCI, including not only pain and motor dysfunction, but also preventingneuronal regeneration. Microglia and macrophages play very important roles in secondary injury. Microglia reside in spinal parenchyma and survey the microenvironment through the signals of injury or infection. Macrophages are derived from monocytes recruited to injured sites from the peripheral circulation. Activated resident microglia and monocyte-derived macrophages induce and magnify immune and inflammatory responses not only by means of their secretory moleculesand phagocytosis, but also through their influence on astrocytes, oligodendrocytes and demyelination. In this review, we focus on the roles of mi- croglia and macrophages in secondary injury and how they contribute to the sequelae of SCI.

Expression of long non-coding RNAs in complete transection spinal cord injury: a transcriptomic analysis

Long non-coding RNAs(lncRNAs)are abundantly expressed in the central nervous system and exert a critical role in gene regulation via multiple biological processes.To uncover the functional significance and molecular mechanisms of lncRNAs in spinal cord injury(SCI),the expression signatures of lncRNAs were profiled using RNA sequencing(RNA-seq)technology in a Sprague-Dawley rat model of the 10th thoracic vertebra complete transection SCI.Results showed that 116 of 14,802 detected lncRNAs were differentially expressed,among which 16—including eight up-regulated(H19,Vof16,Hmox2-ps1,LOC100910973,Ybx1-ps3,Nnat,Gcgr,LOC680254)and eight down-regulated(Rmrp,Terc,Ngrn,Ppp2r2b,Cox6a2,Rpl37a-ps1,LOC360231,Rpph1)—demonstrated fold changes>2 in response to transection SCI.A subset of these RNA-seq results was validated by quantitative real-time PCR.The levels of 821 mRNAs were also significantly altered post-SCI;592 mRNAs were up-regulated and 229 mRNAs were down-regulated by more than 2-fold.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses showed that differentially expressed mRNAs were related to GO biological processes and molecular functions such as injury and inflammation response,wound repair,and apoptosis,and were significantly enriched in 15 KEGG pathways,including cell phagocytosis,tumor necrosis factor alpha pathway,and leukocyte migration.Our results reveal the expression profiles of lncRNAs and mRNAs in the rat spinal cord of a complete transection model,and these differentially expressed lncRNAs and mRNAs represent potential novel targets for SCI treatment.We suggest that lncRNAs may play an important role in the early immuno-inflammatory response after spinal cord injury.This study was approved by the Administration Committee of Experimental Animals,Guangdong Province,China.

Time representation of mitochondrial morphology and function after acute spinal cord injury

Changes in mitochondrial morphology and function play an important role in secondary damage after acute spinal cord injury. We recorded the time representation of mitochondrial morphology and function in rats with acute spinal cord injury. Results showed that mitochondria had an irregular shape, and increased in size. Mitochondrial cristae were disordered and mitochondrial membrane rupture was visible at 2–24 hours after injury. Fusion protein mitofusin 1 expression gradually increased, peaked at 8 hours after injury, and then decreased to its lowest level at 24 hours. Expression of dynamin-related protein 1, amitochondrial fission protein, showed the opposite kinetics. At 2–24 hours after acute spinal cord injury, malondialdehyde content, cytochrome c levels and caspase-3 expression were increased, but glutathione content, adenosine triphosphate content, Na＋-K＋-ATPase activity and mitochondrial membrane potential were gradually reduced. Furthermore, mitochondrial morphology altered during the acute stage of spinal cord injury. Fusion was important within the first 8 hours, but fission played a key role at 24 hours. Oxidative stress was inhibited, biological productivity was diminished, and mitochondrial membrane potential and permeability were reduced in the acute stage of injury. In summary, mitochondrial apoptosis is activated when the time of spinal cord injury is prolonged.

Topiramate as a neuroprotective agent in a rat model of spinal cord injury

Topiramate（TPM） is a widely used antiepileptic and antimigraine agent which has been shown to exert neuroprotective effects in various experimental traumatic brain injury and stroke models. However, its utility in spinal cord injury has not been studied extensively. Thus, we evaluated effects of TPM on secondary cellular injury mechanisms in an experimental rat model of traumatic spinal cord injury（SCI）. After rat models of thoracic contusive SCI were established by free weight-drop method, TPM（40 mg/kg） was given at 12-hour intervals for four times orally. Post TPM treatment, malondialdehyde and protein carbonyl levels were significantly reduced and reduced glutathione levels were increased, while immunoreactivity for endothelial nitric oxide synthase, inducible nitric oxide synthase, and apoptotic peptidase activating factor 1 was diminished in SCI rats. In addition, TPM treatment improved the functional recovery of SCI rats. This study suggests that administration of TPM exerts neuroprotective effects on SCI.

Apoptosis and P53 protein expression after rat spinal cord injury

objective: To determine the DNA damage, neuronal and glial cells apoptosis and expression of P53 protein in the rat spinal cord after compression injury. Methods: Terminal deoxynucleotidyl transferase . mediated dUPT nick end labeling (TUNEL), DNA gel electrophoresis and immunohistochemistry techniques were used to detect DNA fragmentation in the injured rat spinal cord. Results: The apoptosic cells and P53 protein presented at 4 h after spinal cord injury with a maximum presence at 24 h in the injuried T8.9 DNA fragmentation presented typical ladder pattern on agarose gel at 24 h. Conclusion: There are lots of neuronal and glial cells apoptosis with DNA damage after SCl. The P53 protein may play an important role in induc tion of neuronal and glial cells to apoptosis.

Effects of low dose Glibenclamide on secondary damage after acute spinal cord injury in rats

Objective To investigate the effects of Glibenclamide on reduction of secondary damage after acute spinal cord injury in rats.Methods Ninety rats were randomly divided into control group(laminectomy alone),spinal cord injury group(injury group),and treatment group(treated

Radiation exposure and reduction in the operating room: Perspectives and future directions in spine surgery

Intraoperative imaging is vital for accurate placement of instrumentation in spine surgery. However, the use of biplanar fluoroscopy and other intraoperative imaging modalities is associated with the risk of significant radiation exposure in the patient, surgeon, and surgical staff. Radiation exposure in the form of ionizing radiation can lead to cellular damage via the induction of DNA lesions and the production of reactive oxygen species. These effects often result in cell death or genomic instability, leading to various radiation-associated pathologies including an increased risk of malignancy. In attempts to reduce radiation-associated health risks, radiation safety has become an important topic in the medical field. All practitioners, regardless of practice setting, can practice radiation safety techniques including shielding and distance to reduce radiation exposure. Additionally, optimization of fluoroscopic settings and techniques can be used as an effective method of radiation dose reduction. New imaging modalities and spinal navigation systems have also been developed in an effort to replace conventional fluoroscopy and reduce radiation doses. These modalities include Isocentric Three-Dimensional C-Arms, O-Arms, and intraoperative magnetic resonance imaging. While this influx of new technology has advanced radiation safety within the field of spine surgery, more work is still required to overcome specific limitations involving increased costs and inadequate training.

Clemastine in remyelination and protection of neurons and skeletal muscle after spinal cord injury

Spinal cord injuries affect nearly five to ten individuals per million every year. Spinal cord injury causes damage to the nerves, muscles, and the tissue surrounding the spinal cord. Depending on the severity, spinal injuries are linked to degeneration of axons and myelin, resulting in neuronal impairment and skeletal muscle weakness and atrophy. The protection of neurons and promotion of myelin regeneration during spinal cord injury is important for recovery of function following spinal cord injury. Current treatments have little to no effect on spinal cord injury and neurogenic muscle loss. Clemastine, an Food and Drug Administration-approved antihistamine drug, reduces inflammation, protects cells, promotes remyelination, and preserves myelin integrity. Recent clinical evidence suggests that clemastine can decrease the loss of axons after spinal cord injury, stimulating the differentiation of oligodendrocyte progenitor cells into mature oligodendrocytes that are capable of myelination. While clemastine can aid not only in the remyelination and preservation of myelin sheath integrity, it also protects neurons. However, its role in neurogenic muscle loss remains unclear. This review discusses the pathophysiology of spinal cord injury, and the role of clemastine in the protection of neurons, myelin, and axons as well as attenuation of skeletal muscle loss following spinal cord injury.

Role of inflammation in neurological damage and regeneration following spinal cord injury and its therapeutic implications

Spinal cord injury(SCI)is an incurable trauma that frequently results in partial or complete loss of motor and sensory function.Massive neurons are damaged after the initial mechanical insult.Secondary injuries,which are triggered by immunological and inflammatory responses,also result in neuronal loss and axon retraction.This results in defects in the neural circuit and a deficiency in the processing of information.Although inflammatory responses are necessary for spinal cord recovery,conflicting evidence of their contributions to specific biological processes have made it difficult to define the specific role of inflammation in SCI.This review summarizes our understanding of the complex role of inflammation in neural circuit events following SCI,such as cell death,axon regeneration and neural remodeling.We also review the drugs that regulate immune responses and inflammation in the treatment of SCI and discuss the roles of these drugs in the modulation of neural circuits.Finally,we provide evidence about the critical role of inflammation in facilitating spinal cord neural circuit regeneration in zebrafish,an animal model with robust regenerative capacity,to provide insights into the regeneration of the mammalian central nervous system.

神经妥乐平对兔脊髓放射性损伤的治疗作用

目的:观察神经妥乐平(NTP)对兔放射性脊髓损伤的治疗效果.方法:新西兰大白兔60只,体重2.5～3kg,随机分成实验组与对照组,每组30只,各组再随机分成5小组接受不同剂量的放射性照射,每组6只,照射剂量分别为20Gy,25Gy,35Gy,40Gy,45Gy.照射部位为T4水平,所有兔在照射前以及照射后均做MRI检查.兔出现临床表现后实验组每天肌肉注射NTP 3.6NU一次,共4周;对照组每天肌肉注射生理盐水3ml一次,共4周.观察两组治疗前后兔症状变化和脊髓MRI的表现及病理改变.结果:照射后平均16周时,动物受照射脊髓节段MRI出现明显异常信号,而临床症状的出现约在照射后平均18周.在治疗结束后3周,实验组各项指标(瘫痪、尿潴留和针刺反应)与对照组相比均明显改善(P＜0.05);MRI图像上实验组治疗后病灶面积比治疗前明显缩小:神经元肿胀明显减轻,髓鞘结构趋于正常.而对照组变化不明显.结论:神经妥乐平对放射性脊髓损伤有一定的治疗作用.

金匮肾气丸对大鼠脊髓近距离放射损伤细胞凋亡形态学的影响

目的观察金匮肾气丸对192Ir近距离照射大鼠脊髓后细胞凋亡的影响。方法将120只SD大鼠随机分成为模型组、金匮肾气丸组、泼尼松组和正常组,各组每日给予相应的药物,连续给药14天后,除正常组外,其它各组均采用后装技术在大鼠脊髓进行192Ir近距离照射,照射剂量均为22Gy,大鼠分别在照射后8h、24h、4周时取损伤节段脊髓,应用HE、TUNEL染色,光镜和电镜观察。结果近距离照射各组大鼠脊髓后8h、24h时HE染色均未观察到明显组织结构上的改变,4周时脊髓白质区出现组织疏松和出血等病理改变。TUNEL染色和电镜显示:与模型组比较,金匮肾气丸组和泼尼松组照射后8h大鼠脊髓的细胞凋亡指数显著下降(P<0·01),而在照射后24h、4周时间点则差异无显著性。结论金匮肾气丸具有肾上腺皮质激素样作用,可对抗大鼠近距离放射性损伤早期胶质细胞的凋亡作用。

鼻咽癌放疗后放射性脑干和脊髓损伤的磁共振表现

背景与目的:鼻咽癌(nasopharyngealcarcinoma,NPC)放射治疗后脑干和颈髓放射性损伤的诊断相当重要。磁共振(magneticresonanceimaging,MRI)用于诊断鼻咽癌放疗后放射性脑损伤已有较多报道,但脑干和颈髓放射性损伤的MRI表现文献报道较少。本研究旨在分析鼻咽癌放疗后脑干和颈髓放射性损伤的MRI特征。方法:对60例鼻咽癌患者在放射治疗后6个月至5年内进行了MRI检查,MRI检查序列包括T1-weightedimage(T1WI)、T2-weightedimage(T2WI)、fluidattenuatedinversionrecovery(FLAIR);所有患者均做了MRI增强扫描。结果:6例为颈髓放射性损伤;54例为脑干放射性损伤,其中脑桥20例,桥脑基底部与延髓上段26例,中脑受累3例,延髓5例。病灶在MRI上T1WI表现为等信号和低信号,T2WI上为高信号,增强后病灶无强化者11例(18.3%),强化者49例(81.7%)。强化者有2种强化形式:均匀斑片状强化(21例,42.9%)和不均匀的既有环形又有斑片状的强化(28例,57.1%)。结论:MRI可以清晰地显示脑干和脊髓放射性损伤的病灶,结合病史可以确诊。