A further development in spinal instrumentation

Summary The purpose of this paper is to describe the concepts and use of a new, versatile instrumentation system, the universal spine system (USS), that has been designed to have wide application for pathology of the thoracolumbar spine. Many instrumentation systems for thoracolumbar spinal surgery now exist that were each originally designed to address specific areas of spinal pathology. For example, the recent systems designed to treat scoliosis deformities do not provide the instrumentation and implant support to adequately address other spinal disorders, such as fractures. In addition, most posterior implant systems were not designed for use anteriorly, and vice versa. As a result, surgeons have been required to become familiar with several different instrumentation systems in order to meet the varied needs of a spinal surgery practice. The objective of designing a new system was to simplify the surgical treatment of the commonly encountered spinal disorders by providing the surgeon with a single set of instruments and implants that could be used to treat tumors, trauma, deformities, and degnerative conditions affecting the thoracolumbar spine, via either an anterior or a posterior approach. This paper describes the development of the concepts, instruments, and implants of the USS and provides examples of its application in several case illustrations.     

Spinal arthrodesis for spinal deformity using posterior instrumentation and sublaminar wiring

Summary One hundred consecutive patients with spinal deformity due to various diagnoses were treated by posterior spinal arthrodesis with instrumentation and multiple sublaminar wires. Both the Harrington and Luque rodding systems were used. A total of 1128 wire loops were passed. No patient developed paraparesis or paraplegia, but three had transient sensory disturbance. There were no cases of broken rods or wires. Forty-five of the patients had no postoperative support. This worked well for neuromuscular scolioses, but for idiopathic scoliosis there was a disturbing loss of correction in many cases. Fusion to the sacrum was best accomplished with the Galveston technique, other methods having a high rate of pseudarthrosis. The main benefits appeared to be the ability to stabilize neuromuscular patients without the use of external immbolization, and the correction of thoracic lordosis.Supported in part by the Medical Education and Research Association of Gillette Children's Hospital Read in part at: Scoliosis Research Society, Orlando, Florida, September, 1984. S.I.C.O.T., London, England, October, 1984     

Anterior thoracoscopic surgery followed by posterior instrumentation and fusion in spinal deformity

Many authors believe thoracoscopic surgery is associated with a lower level of morbidity compared to thoracotomy, for anterior release or growth arrest in spinal deformity. Others believe that anterior release achieved thoracoscopically is not as effective as that achieved with the open procedure. We evaluated the clinical results, radiological correction and morbidity following anterior thoracoscopic surgery followed by posterior instrumentation and fusion, to see whether there is any evidence for either of these beliefs. Twenty-nine patients undergoing thoracoscopic anterior release or growth arrest followed by posterior fusion and instrumentation were evaluated from a clinical and radiological viewpoint. The mean follow-up was 2 years (range 1–4 years). The average age was 16 years (range 5–26 years). The following diagnoses were present: idiopathic scoliosis (n = 17), neuromuscular scoliosis (n = 2), congenital scoliosis (n = 1), thoracic hyperkyphosis (n = 9). All patients were satisfied with cosmesis following surgery. Twenty scoliosis patients had a mean preoperative Cobb angle of 65.1° (range 42°–94°) for the major curve, with an average flexibility of 34.5% (42.7°). Post operative correction to 31.5° (50.9%) and 34.4° (47.1%) at maximal follow-up was noted. For nine patients with thoracic hyperkyphosis, the Cobb angle averaged 81° (range 65°–96°), with hyperextension films showing an average correction to 65°. Postoperative correction to an average of 58.6° was maintained at 59.5° at maximal follow-up. The average number of released levels was 5.1 (range 3–7) and the average duration of the thoracoscopic procedure was 188 min (range 120–280 min). There was a decrease in this length of time as the series progressed. No neurologic or vascular complications occurred. Postoperative complications included four recurrent pneumothoraces, one surgical emphysema, and one respiratory infection. Thoracoscopic anterior surgery appears a safe and effective technique for the treatment of paediatric and adolescent spinal deformity. A randomised controlled trial, comparing open with thoracoscopic methods, is required. Received: 11 October 1999 Revised: 20 April 2000 Accepted: 16 May 2000     

脊柱内固定术后翻修原因及其治疗对策

目的:分析脊柱内固定翻修的原因并探讨翻修手术策略。方法:对我院自2004年1月～2011年12月收治的行脊柱内固定翻修手术的44例患者资料进行回顾总结,平均随访3年(1～81个月)。翻修原因可分为:(1)内置物相关副损伤,3例;(2)内置物断裂、移位,21例;(3)内置物位置欠佳,3例;(4)内置物丧失作用,1例;(5)内置物残留,1例;(6)内置物邻近节段退变,2例;(7)内置物影响感染控制,12例;(8)血肿致神经压迫,1例。针对不同原因采取相应的翻修策略,对内置物进行了更换或拆除。所有内固定翻修患者采用X线平片、三维CT、MRI进行影像学评价,其中脊髓型颈椎病患者采用改良JOA评分法进行评价,腰椎退变性疾病患者采用Stucki评分法进行评价,脊柱骨折脱位伴脊髓损伤的病例采用ASIA损伤分级进行评价。结果:所有翻修手术均顺利完成,无术中并发症发生。1例脊髓型颈椎病患者首次行前路减压内固定手术后出现血肿压迫脊髓,翻修手术行血肿清除,术后再次出现血肿压迫,造成短暂的神经功能障碍,再次翻修取出内固定物及清除血肿后,患者神经症状恢复。脊髓型颈椎病患者翻修术前JOA评分为17.38分,术后为17.46分,手术前后无明显变化;腰椎退变性疾病患者根据Stucki评分90%的患者对翻修手术满意,90%的患者翻修术后疼痛缓解并对行走功能恢复满意,80%的患者对下肢力量及平衡能力满意;脊柱骨折脱位伴脊髓损伤患者翻修手术后ASIA损伤分级无变化。所有患者翻修后保留或更换的内固定物位置良好,骨融合率100%,感染得到控制。结论:脊柱内固定术后翻修原因较多,选择合理的翻修手术仍可取得较满意结果。应掌握脊柱内固定应用原理,规范操作以避免翻修手术。     

类泛素化修饰在脊髓损伤中作用的研究进展

脊髓损伤临床常见且预后大多较差,损伤后病理过程相当复杂。近年来,神经系统疾病中类泛素修饰蛋白(SUMO)的作用越来越受到关注。SUMO是一类最新发现的蛋白质翻译后修饰蛋白,类泛素化修饰在人体中多个生理和病理生理学活动过程中发挥重要作用。研究结果显示脊髓损伤后脊髓水肿、脊髓缺血/再灌注、氧化应激和炎性反应等过程中多种重要的蛋白分子均存在SUMO化修饰调控机制,SUMO相关蛋白的应用亦被证明可调节氧化应激延缓神经元的死亡,其与抗氧化剂等其他脊髓治疗性化合物联合使用有望用于预防缺血性损伤的神经保护,本文拟对脊髓损伤过程中有关SUMO化修饰的最新研究作一综述。     

前路减压钛网植骨融合内固定治疗胸腰段脊柱骨折合并脊髓损伤

目的 探讨前路减压钛网植骨融合内固定术治疗单节段胸腰段脊柱骨折合并脊髓损伤的临床效果.方法 自2004年5月至2007年11月应用前路减压钛网植骨融合内固定术治疗63例单节段胸腰段脊柱骨折合并脊髓损伤患者,应用ASIA脊髓损伤神经功能评定法评价患者手术前、后的神经功能恢复情况.结果全部病例均获得至少1年随访,至末次复查时,全部病例均获得植骨融合,内固定物无松动,胸腰段脊柱生理弯曲得以维持,无继发后凸畸形及椎管狭窄.与术前比较,患者术后A-SIA脊髓神经损伤分级明显改善,脊髓神经感觉(t=2.759,P=0.025)、运动(t=4.616,P<0.05)功能评分均显著提高,差异有统计学意义.结论 采用前路减压钛网植骨融合内固定治疗单节段胸腰段脊柱骨折具有减压彻底、神经功能改善率高、一期内固定植骨融合成功率高、脊柱稳定性好等优点,是治疗胸腰段脊柱骨折的有效方法.     

儿童无放射学影像异常的脊髓损伤14例临床分析

目的：提高对儿童无放射学影像异常的脊髓损伤（spinal cord injury without radiographic abnormality，SCIWORA）的认识。方法：对1992年1月至2005年8月北京儿童医院住院患者中诊断为“急性脊髓炎”或“脊髓损伤”的187例进行回顾性分析，发现其中14例为SCIWORA。男6例，女8例。年龄1岁3个月～15岁（15岁1例，1岁3个月～7岁13例）。均有明确外伤史。损伤水平在C6～T9，颈髓2例，胸髓12例。脊髓功能ASIA分级：A级9例，C级4例，D级1例。脊柱X线片和CT未见骨折及脱位；MRI检查示脊髓水肿9例,脊髓出血2例，脊髓挫伤3例。采用综合治疗，包括卧床、局部制动，应用激素、脱水剂、神经营养药等。结果：4例患者失访，10例患者随访6个月～5年，平均2年3个月，4例（C级3例，D级1例）患者（均为MRI显示水肿局限者）在伤后6个月运动、感觉及括约肌功能完全恢复（E级）；6例（A级）患者运动、感觉及括约肌障碍未恢复。2例（A级）病初MRI表现为脊髓弥漫水肿者，伤后4～6个月复查MRI脊髓呈萎缩样改变。结论：对怀疑SCIWORA的儿童，应首选MRI检查。脊髓水肿局限者预后好，水肿弥漫或伴有出血、挫伤者预后差。     

脊髓干细胞移植对脊髓损伤后神经功能的影响

目的探讨胚胎脊髓神经干细胞移植对大鼠脊髓损伤后神经功能恢复的意义。方法160只SD大鼠随机分为空白组,假手术组,脊髓损伤组,细胞移植组,分别在细胞移植后1、2、4周应用斜板实验和Tarlov评分对脊髓损伤后功能恢复进行评价,应用nestin标记观察移植后干细胞的存活情况。结果移植后1周、2周、4周,移植组和对照组斜板试验结果分别为(38．30±0．84)°、(18．50±0．76)°；jm(39．40±0．78)°、(19．70±0．66)°；(45．00±0．81)°、(22．30±0．69)°；Tarlov评分分别为3．37±0．45、2．32±0．34；3．45±0．38、2．41±0．43；3．63±0．47、2．45±0．48；有统计学意义(P＜0．01),免疫组织化学观察可见在损伤的脊髓组织中有神经干细胞的存活。结论胚胎脊髓干细胞移植对脊髓损伤后神经功能恢复有促进作用。     

Longitudinal changes in medical complications in adults with pediatric-onset spinal cord injury

Objectives

To determine longitudinal changes in the occurrence of medical complications in adults with pediatric-onset spinal cord injury (SCI).

Design

Longitudinal study of long-term outcomes.

Setting

Community.

Participants

Individuals who had sustained an SCI before age 19, were 23 years of age or older at initial interview, and followed annually between 1996 and 2011. They were classified into four American Spinal Injury Association (ASIA) Impairment Scale (AIS) severity groups: C1–4 AIS ABC, C5–8 AIS ABC, T1–S5 AIS ABC, AIS D.

Outcome measures

Generalized estimating equation (GEE) models were formulated to obtain the odds ratio (OR) of having a medical complication over time.

Results

A total of 1793 interviews were conducted among 226 men and 125 women (86% Caucasian; age at baseline, 26.7 ± 3.6 years; time since injury at baseline, 12.9 ± 5.2 years). Odds of complication occurrence over time varied among severity groups, with increased ORs of severe urinary tract infection (1.05, confidence interval (CI) 1.02–1.09), autonomic dysreflexia (AD) (1.09, CI 1.05–1.14), spasticity (1.06, CI 1.01–1.11), pneumonia/respiratory failure (1.09, CI 1.03–1.16), and hypertension/cardiac disease (1.07, CI 1.01–1.15) in the C1-4 ABC group; AD (1.08, CI 1.04–1.13) and pneumonia/respiratory failure (1.09, CI 1.02–1.16) in the C5–8 ABC group; and hypertension/cardiac disease (1.08, CI 1.02–1.14) in the T1–S5 ABC group. Upper extremity joint pain had increased odds of occurrence in all injury severity groups.

Conclusion

The significantly increased odds of having medical complications over time warrants awareness of risk factors and implementation of preventive measures to avoid adverse consequences of complications and to maintain independence in individuals with pediatric-onset SCI.     

Mechanical consequences of rod contouring and residual scoliosis in sublaminar segmental instrumentation

The mechanical performance of contoured Luque rods in a neuromuscular model of spine deformity was examined to define an upper limit of deformity above which rod stresses would exceed the endurance limit for 316L stainless steel and therefore predict fatigue failure. Bovine constructs varying from 0-120 degrees scoliosis were loaded axially, with strain recordings obtained at the apex of the curve. Relatively low loads produced enough tensile stress to contemplate implant fatigue in all except the nondeformed (0 degrees) construct. Construct stiffness was found to decrease rapidly in spines with greater than 38 degrees deformity. In addition, data on patients who had suffered rod fracture from four different centers were found to compare favorably with experimental observations. We conclude that the vulnerability of Luque rod constructs to implant failure, from a mechanical standpoint, is greater than is generally assumed. Cross-linking of rods was found to increase stiffness. Methods to decrease tensile stresses in the implants and increase stiffness include external immobilization, larger diameter rods, and procedures to enhance correction.     

人胚神经干细胞移植治疗大鼠脊髓损伤

目的 探讨人胚神经干细胞（hNSC）移植治疗脊髓损伤（SCI）的可行性。方法 分离、培养和鉴定hNSC；用5溴-2脱氧尿苷嘧啶（BrdU）标记hNSC，并将其移植到14只T10半横断的Wistar大鼠损伤脊髓内（另外14只T10半横断损伤的大鼠作为对照组，仅损伤脊髓内注射DMEM／F12培养液），用BrdU的FITC免疫荧光染色检测移植细胞的存活和迁徙，用NF-200、GFAP免疫组织化学鉴定移植细胞的分化，BBB评分评定大鼠功能恢复情况。结果 （1）获得了大量的hNSC；（2）用免疫组织化学可以检测到移植的hNSC能在体内长时间存活（达2个月）并向远处迁徙，并分化为神经元和胶质细胞；（3）检测到实验组大鼠BBB得分明显高于对照组大鼠（P〈0．01），在SCI后第10周时实验组和对照组BBB得分最大差距达到2．1分。结论 hNSC移植能促进SCI大鼠后肢功能恢复，它是SCI移植治疗较有价值的细胞资源。     

应用改进封闭脊髓窗技术活体观察脊髓损伤后脊髓微循环变化

目的:探讨应用改进的封闭脊髓窗技术活体观察大鼠脊髓损伤后脊髓微循环变化的可行性及效果.方法:改进传统的脊髓窗,设计带有打击探头的脊髓窗.45只SD大鼠随机分为对照组(n=20)和实验组(n=25).实验组大鼠脊髓窗安装完成后进行窗内打击脊髓,然后即时观察并记录打击后2h内打击点周围微动脉直径的变化.对照组大鼠在脊髓窗安装完成后不进行打击,只对脊髓微动脉直径进行连续2h观察并记录.术后两组大鼠进行BBB运动功能评分,处死动物取脊髓标本切片,HE染色,观察脊髓组织改变情况.结果:实验组大鼠打击点周围2～6mm的微动脉直径在打击后10min、30min、1h、2h各时间点均较打击前明显减小,术后2d及7d时BBB功能评分明显低于术前,病理切片可见脊髓打击区神经组织变性液化.对照组大鼠在观测开始及结束时脊髓微动脉直径无明显变化,术后BBB评分与术前BBB评分均为21分,病理切片未见脊髓损伤表现.结论:改进的封闭脊髓窗技术可以有效地实现窗内打击脊髓,并可以安全地对脊髓表面微血管进行活体观测.脊髓表面打击点周围2～6mm的微动脉在脊髓损伤后10min即发生痉挛.     

TRH对大鼠脊髓损伤后SCBF和SEP的影响

脊髓损伤（ＳＣＩ）后内源性阿片肽释放，并参与脊髓的继发损伤机制。ＴＲＨ可阻断阿片肽的自主神经效应，而不影响痛觉。本实验探讨大剂量ＴＲＨ（２ｍｇ／ｋｇ／ｈ）治疗对大鼠脊髓打击伤（Ａｌｌｅｎｓ法１０ｇｘ５ｃｍ）后脊髓血流量（ＳＣＢＦ）和脊髓诱发电位（ＳＥＰ）的影响。脊髓损伤后１ｈ，ＳＣＢＦ开始显著下降，持续至伤后２４ｈ，ＳＥＰ峰潜时呈进行性延长趋势；伤后即刻静脉注射ＴＲＨ（２ｍｇ／ｋｇ／ｈ，共５次），可使伤后即刻和２４ｈ的ＳＣＢＦ显著升高，并使伤后ＳＣＢＦ下降时间延迟３ｈ，同时ＳＥＰ峰潜时有不同程度改善。结果表明，ＴＲＨ对受伤脊髓早期有一定的防治作用，并具有一定的后发效应；同时也可促进脊髓的神经传导功能。本文亦对ＴＲＨ治疗ＳＣＩ的病理生物学机制进行了讨论。     

神经生物膜介导神经干细胞联合雪旺细胞移植治疗脊髓损伤

目的探讨以壳聚糖-胶原偶联生物膜为载体，联合移植雪旺细胞（SCs）与神经干细胞（NSCs）在治疗脊髓损伤中的作用，并评价该方法的疗效。方法大鼠孕鼠体内取出胎鼠分离获得原代NSCs进行体外培养，大鼠乳鼠坐骨神经中分离获得原代SCs进行体外培养，分别传代4代获得大量细胞。40只Wistar大鼠建立大鼠脊髓半横断动物模型并随机分为4组：空白对照组，SCs移植组，NSCs移植组，NSCs联合SCs移植组。将胎鼠NSCs和乳鼠SCs共同种植于胶原-壳聚糖偶联的神经生物膜上，移植入大鼠脊髓半横断模型的脊髓损伤部位，对动物进行BBB脊髓功能评分，BDA神经示踪标记，标记2周后处死动物取出动物脊髓组织进行Cy3荧光素染色，p75免疫荧光染色及脊髓组织苏木素-伊红（HE）染色。结果4组实验动物BBB评分组间比较差异有统计学意义（ANOVA，P〈0．05），p75免疫荧光染色阳性物面积比较各组之间差异有统计学意义（ANOVA，P〈0．05）。结论NSCs和SCs能够在胶原-壳聚糖偶联的神经生物膜上共同生长分化，并且SCs能够诱导NSCs产生轴突定向生长。壳聚糖-胶原生物膜介导的SCs联和NSCs移植治疗脊髓损伤可使神经部分再通。     

完全性脊髓损伤早期患者脑皮层结构变化的磁共振研究

目的:观察完全性脊髓损伤(SCI)患者在损伤早期大脑皮层结构是否存在改变及其与患者感觉运动功能损伤程度的相关性。方法:2013年10月~2013年12月在北京军区总医院骨科住院的13例完全性SCI患者被纳入研究,其中男7例,女6例,年龄26.3±7.9岁(21~40岁)。同时招募健康对照15例,其中男8例,女7例,年龄25.7±5.8岁(20~38岁)。SCI患者损伤时间为64.4±20.3d(28~84d)。13例患者脊髓功能用美国脊髓损伤协会(ASIA)分级标准评定均为A级,其中ASIA运动评分为36.5±25.9(18~55)分,针刺觉评分为43.3±27.3(27~88)分,轻触觉评分为42.5±30.3(25~80)分。所有SCI患者在入院当天接受结构MRI检查。MRI扫描后的结构数据用基于体素形态学分析的方法(VBM)来分析,对比分析13例完全性SCI患者和15例年龄、性别相匹配的健康对照者大脑灰质体积改变的区域。运用Person相关性分析探讨脑皮层结构改变与患者运动感觉功能之间的关系。结果:与健康对照者相比,完全性SCI患者初级运动中枢、初级躯体感觉中枢和辅助运动区存在明显的灰质萎缩(P0.05)。相关性分析显示完全性SCI患者上述区域萎缩程度与患者的ASIA运动评分以及ASIA感觉评分无明显相关性(P0.05)。结论:在损伤早期,完全性SCI患者初级运动中枢、初级躯体感觉中枢和辅助运动区存在明显的灰质萎缩,上述区域萎缩程度与患者的ASIA评分无明显相关性。     

盐酸戊乙奎醚对大鼠急性脊髓损伤修复的作用

目的 观察盐酸戊乙李醚对大鼠急性脊髓损伤后脊髓损伤修复的作用.方法 选用健康成年雄性SD大鼠90只分为3组,A组:盐酸戊乙奎醚治疗组(n=40),从造模后1 h开始,经腹腔注射盐酸戊乙奎醚每天3 mg/kg,卣至取材;B组:单纯损伤组(n=40),单纯损伤组同法给予等量生理盐水;C组:正常对照组(n=10),不作任何处理.A组、B组大鼠采用改良Allen'S重物坠落法在T10段制作急性脊髓损伤模型.各组取材前24 h腹腔注射溴脱氧尿嘧啶核苷(BrdU)溶液,对距离损伤中心5 mm处的脊髓进行BrdU、nestin阳性细胞数检测.结果 C组脊髓中央管周围及外膜可见少量BrdU阳性细胞,几乎看不到nestin阳性细胞.B组造模后24 h即可见大量BrdU阳性细胞,1周达到高峰,2周后开始明显减少,4周时仅见少量BrdU阳性细胞.与B组比较,24 h时A组BrdU阳性细胞差异无统计学意义,1周时BrdU阳性细胞数明显增多(P<0.05),至2周时仍处于较高水平,4周时仍有大量BrdU阳性细胞表达.B组造模后7 d nestin阳性细胞增多,14 d达到高峰,28 d时可见极少量nestin瞄H性细胞.与B组比较,7 d时A组nestin阳性细胞差异无统计学意义,14 d时nestin阳性细胞数明显增多,至28 d时仍处于较高水平(P<0.05).结论 盐酸戊乙奎醚能够促进急性脊髓损伤大鼠损伤区BrdU及nestin的表达,提示有促进神经干细胞增殖的能力,增强脊髓损伤后自体的修复功能.     

Adult spinal cord injury with no radiographic bone lesion: analysis of nine consecutive patients

Background

Spinal cord injury with no radiographic bone lesion described as spinal cord injury without radiographic abnormality (SCIWORA) in childhood is less often reported in adults than in children. This study was undertaken to report our experience in the management of nine cases over 25 years.

Patients and methods

This was a retrospective study from 1985 to 2009 concerning nine adult patients who sustained spinal cord injury with no radiographic abnormality. The ratio among all cervical spine traumas for the same period was 2.21%. Magnetic resonance imaging (MRI) was performed in all the patients. The patients’ clinical status at the time of admission and discharge was evaluated using the Frankel's grading system. We report the results based on the clinical, epidemiologic and radiological findings and outcomes.

Results

The mean age of our population was 37.43 years, ranging from 18 to 60 years. All the patients were men. The main etiology was falls (5/9) followed by road traffic accidents (4/9). According to the Frankel's grading system, four patients (44.45%) were grade A, four were grade B (44.45%), and one was grade C (11.11%). On MRI, medullar lesions were: contusion, non-compressive cervical disc herniation, cervical spine stenosis, and two cases of normal cervical spine. Four patients were operated on via the posterior cervical spine approach (laminectomy, C3-C7 in three cases and C1-C3 in one case). The other five patients were treated orthopaedically for 6 to 8 weeks. Three patients (3/9), who were Frankel's grade B and C with no demonstrable injury on MRI, improved to Frankel a useful neurological grade (Frankel's grades D or E) at the time of discharge. One patient evaluated as Frankel's grade A died from cardiovascular disturbance.

Conclusion

Spinal cord injury with no radiographic abnormality accounted for 2.21% of cases of spinal cord injury in our series. MRI is the investigation of choice, having diagnostic and prognostic value because it demonstrates neural and extraneural injuries and helps to identify surgically correctable abnormalities.     

Neurological complications in segmental spinal instrumentation: analysis of 750 patients

The authors report on neurological damage caused by the use of sublaminar segmental fixation in the correction of vertebral deformities. Three groups were reviewed: 600 patients instrumented with Harrington rods and segmental wiring, 50 patients treated with the Hartshill system and 100 patients instrumented with Luque bars. All of the patients were operated on using sublaminar wiring fixation. We report two transitory neurological complications among the 600 patients with Harrington rod instrumentation and segmental wiring, two permanent neurological deficits among the 50 cases treated with the Hartshill system and none among the 100 patients instrumented using Luque bars. The purpose of this study is to analyse the causes of these neurological complications, which occurred late in all four of the cases described.Presented at the ESDS meeting, Birmingham, 1994, and selected for full publication     

神经干细胞单细胞与神经球治疗大鼠脊髓损伤的对比研究

目的比较神经干细胞(neural stem cells,NSCs)单细胞与神经球移植治疗大鼠脊髓损伤(spinal cordinjury,SCI)的效果,研究两种NSCs移植方法治疗SCI的有效性。方法取成年SD大鼠2只,体外分离脊髓组织并行NSCs培养,取第3代细胞行Hoechst33342、巢蛋白染色及贴壁分化鉴定。另取成年SD大鼠(体重230～250 g)60只,随机分为3组,每组20只,采用改良Allen法制备大鼠脊髓T10损伤模型。各组分别于SCI处缓慢注射5μL生理盐水(A组)、第3代NSCs单细胞(B组)、第3代NSCs神经球(C组)。分别于术前及术后3、7、14、21、28 d采用BBB行为功能评定量表评定各组大鼠后肢功能;术后各时间点取材行HE染色和微管相关蛋白2(microtubule-associated protein 2,MAP-2)免疫组织荧光染色观察。结果经形态学观察及鉴定,所培养的细胞为NSCs。术后3 d各组BBB评分较术前显著下降,术后3、7 d各组间BBB评分比较差异均无统计学意义(P>0.05);随时间延长BBB评分不同程度增加,术后14、21、28 d,B、C组BBB评分优于A组,C组优于B组,比较差异均有统计学意义(P<0.05)。HE染色示C组较A、B组脊髓结构清晰,瘢痕形成少。MAP-2免疫组织荧光染色示,术后3、7 d各组间阳性细胞数比较差异均无统计学意义(P>0.05);术后14、21、28 d,B、C组阳性细胞数显著多于A组,C组多于B组,差异有统计学意义(P<0.05)。结论采用NSCs神经球移植较单细胞移植更明显促进NSCs向神经元分化,更有利于SCI后下肢功能恢复。