Mechanical compression of the lumbar nerve root alters pain-related behaviors induced by the nucleus pulposus in the rat.

The purpose of this study was to refine a method of nerve-root injury in the rat to produce hyperalgesia, a pain-related behavior, and to determine if there were any relationships between the histological extent of nerve-root injury and the magnitude of hyperalgesia. Three methods were used to produce hyperalgesia: irritation of a nerve root by ectopic nucleus pulposus, silk loop alone, or both silk loop and ectopic nucleus pulposus. Autologous nucleus pulposus obtained from coccygeal intervertebral discs was relocated on the lumbar nerve roots after laminectomy. Two loops of 4-0 silk were placed around the exposed nerve roots. Hyperalgesia was measured preoperatively and postoperatively. The distribution of myelinated axons in the dorsal nerve roots was evaluated histologically. Mechanical hyperalgesia was detected in rats in which autologous nucleus pulposus was applied to the nerve root but not in those in which silk loops were used. Silk loops around the nerve root resulted in thermal hyperalgesia only in rats in which autologous nucleus pulposus was applied to the nerve root. Fewer large myelinated fibers were seen in the rats in which silk loops were used. Although a silk loop around the nerve root was not sufficient to produce hyperalgesia, supplemental application of autologous nucleus pulposus to the nerve root produced thermal hyperalgesia. It is possible that mechanical constriction of the nerve root alters the pain-related behavior elicited by chemical factors from the nucleus pulposus.     

皮层躯体感觉诱发电位在监测腰神经根损伤中作用的研究

目的：利用大鼠髓核突出动物模型。探索皮层躯体感觉诱发电位（CSEP）的波幅和潜伏期变化是否与神经根性疼痛有关系。方法：取大鼠自体尾部的髓核无压迫下放置在L4和L5神经根上，制成髓核突出动物模型。分别在术后3d,1,2及4周观察大鼠术侧肢体机械刺激敏感性和热刺激敏感性和热刺激敏感性的变化，并引出大鼠后肢CSEP，观察术侧肢体CSEP的变化。结果：在无明显机械压迫的情况下，大鼠腰神经根上植入自体髓核可产生痛觉过敏，CSEP波幅增高。结论：髓核自身是引起腰腿痛的重要原因，CSEP波幅的增高与神经根性疼痛有一定相关性。     

一氧化氮在神经根性疼痛中的作用

目的：探索一氧化氮(NO)在髓核突出所致的神经根性疼痛中的作用。方法：取大鼠自体尾椎髓核无压迫下放置在L4和L5神经根表面，分别在术后3d及1、2、3、4周时观察大鼠后足机械刺激和热刺激敏感性的变化，并用免疫组化方法对移植髓核中的一氧化氮合酶(NOS)进行检测．探索NO在疼痛中的作用：结果：在无明显机械压迫情况下，大鼠腰神经根上放置自体髓核可产生痛觉过敏，移植髓核组织中NOS染色阳性一结论：髓核自身是引起腰腿痛的重要原因，NO可能参与疼痛的产生.     

实验性髓核突出诱致神经根性疼痛机理的探讨

目的 探索单纯自体髓核组织放置于神经根邻近,并与神经根接触后对大鼠后爪机械刺激痛觉敏感性的影响。方法 取大鼠自体尾椎髓核组织无压迫下置于腰4和腰5神经根旁并与神经根相接触,术后不同的时间点观察大鼠双侧后爪机械刺激敏感性的变化;同时采用HE染色方法观察髓核组织和神经根的变化。结果 无明显机械压迫情况下,自体髓核组织与神经根接触后可诱导大鼠术侧后爪出现明显的机械性痛觉敏感性的升高;HE染色显示髓核组织发生了明显的炎性改变,邻近神经根出现空泡变性。结论 髓核组织自身介导的炎症反应可能参与机械性痛觉敏感性的发生。除机械因素外,突出髓核组织发生的炎症反应也可能是腰椎间盘突出所致坐骨神经痛的原因。     

大鼠正常与退变性髓核突出导致神经根性疼痛的对比研究

目的探讨正常与退变髓核突出对大鼠疼痛阈值以及背根神经节中TNF-α表达的影响,研究椎间盘退变与神经根性疼痛之间的关系。方法72只大鼠随机分为4组:正常对照组(n=18)、假手术组(n=19)、正常髓核(N-NP)组(n=16)和退变髓核(P-NP)组(n=19)。对P-NP组大鼠利用尾椎椎间盘纤维环穿刺的方法建立椎间盘退变模型。分别取出N-NP组和P-NP组大鼠自体的正常髓核与退变髓核组织,置于手术显露后的腰5左侧神经根处,建立髓核突出致神经根性疼痛动物模型。采用行为学测试的方法分别观察各组大鼠术前1天,术后1、4、7、10、14、21天机械刺激阈值与热刺激阈值的变化;采用免疫组化方法分别检测术后第4、14天各组大鼠背根神经节中TNF-α的表达。结果行尾椎间盘纤维环穿刺后2周,组织学与MRI检查均证实椎间盘组织发生明显退变。对照组和假手术组动物未出现明显的痛觉过敏现象,N-NP组和P-NP组大鼠机械性刺激阈值均显著下降,该痛觉过敏现象持续至术后2周消失;与正常髓核组织相比,退变髓核所致机械性刺激阈值下降程度更为严重。各实验组均未发生热刺激阈值的规律性变化。术后第4、14天对照组和假手术组背根神经节中未见TNF-α明显表达,而正常及退变髓核组TNF-α表达量均显著升高。结论大鼠尾椎纤维环穿刺是建立大鼠椎间盘退变模型的一种有效方法。与正常髓核组织相比,发生退变的髓核组织可导致神经根性疼痛的加重,提示椎间盘退变过程中释放的炎症因子在疼痛的发生机制中可能起到了重要作用。     

临床症状体征与影像学检查分离的腰椎间盘突出症的发生机制研究进展

临床会出现少数症状体征与影像学检查结果不相符的腰椎间盘突出症患者,而单纯用传统的突出髓核直接机械压迫刺激神经根的理论不能解释这种反常的腰椎间盘突出症。腰椎间盘髓核的突出与患者临床症状体征的出现受多因素、多环节的影响,脊神经根的间接性机械压迫与神经根牵张效应为主要因素,而反常症状体征的产生往往与突出的髓核自身位置的迁移、神经系统对信息的传递以及髓核与硬膜囊或神经根的相互作用密切相关。此外,突出的髓核组织所继发的局部微循环、炎症改变,相应节段的骨质增生退变和腰椎应力姿势改变诱发此类反常腰椎间盘突出症患者出现多样性的症状体征。同时,一些患者还存在神经或椎体的先天性发育异常,并可能出现影像学检查上的误诊或漏诊。突出髓核对硬膜囊以及周围神经根之间的确切相互作用机制及其继发的局部病理生理、生物力学改变,病变责任节段的确定以及如何克服影像学检查的局限性需进一步研究。     

周围神经旁移植自体髓核对大鼠疼痛行为的影响

目的　探讨髓核在坐骨神经痛发病机制中的作用。方法　选择 16只SD雄性大鼠随机分为两组 ,分别将自体尾椎髓核混悬液注射到腰椎硬膜外腔或坐骨神经周围。测定其后肢机械刺激缩爪阈值。结果　在没有明显机械压迫下 ,腰椎硬膜外腔的髓核使大鼠后肢产生明显的痛觉过敏反应 ,坐骨神经旁的髓核不能使大鼠产生上述变化。结论　与髓核相关的炎症的介质是引起坐骨神经痛的原因之一 ,神经根缺乏神经外膜保护是引起疼痛的解剖学基础。     

经皮穿刺腰椎间盘激光汽化减压术的临床应用

目的：探讨经皮穿刺腰椎间盘激光汽化减压术的临床应用和适应证。方法：采用脊柱后外侧人路，在C形臂X线机引导下，将穿刺针定位于椎间隙中心稍偏后部，插入光导纤维，通过激光脉冲汽化部分髓核组织，降低椎间盘内压力，解除突出物对神经根和硬膜的压迫。结果：本组共治疗116例；131个间隙。参照MacNab疗效评价，优90例，占77．6％；良15例，占12．9％；差9例，其中3例行二次激光汽化治疗，5例改为开放手术。结论：本方法是一种有限治疗，但创伤小，安全有效，对脊柱稳定性没有影响。适用于单纯的包容性腰椎间盘突出症患者。     

髓核对大鼠腰神经根非压迫性损伤的组织形态学研究

目的　观察硬膜外移植自体髓核后 ,大鼠神经根组织形态学的变化。方法　 2 8只Sprague Dawley(SD)雄性大鼠随机分成对照组和实验组 ,分别将自体肌肉混悬液和尾椎髓核混悬液注射到腰椎硬膜外腔 ,对神经根组织进行形态学观察。结果　在无机械压迫情况下 ,大鼠硬膜外移植自体髓核能使马尾神经根组织形态产生明显改变。结论　髓核所致的炎性反应是引起神经根损伤和坐骨神经痛的重要原因之一     

Stable mechanical environments created by a low-tension traction device is beneficial for the regeneration and repair of degenerated intervertebral discs

BACKGROUNDBy blocking the cascade of reactions leading to intervertebral disc degeneration through immobilization-traction, a delay in intervertebral disc degeneration and its regeneration, to some extent, has been observed. However, the precise balance of regulation of the microenvironment of intervertebral disc biomechanics and coordination of the complex spatiotemporal reconstruction of the extracellular matrix have not yet been solved, and clinical results are far from successful.PURPOSEIn the present study, a mechanical degeneration model was constructed to evaluate the possibility and effectiveness of disc regeneration or repair through low-tension traction of degenerated discs so as to provide basic biomechanical information for clinical optimization of the traction device and to establish traction parameters for prevention and treatment of disc degeneration.STUDY DESIGNA macro-, micro-, and nano-level structural analysis of degenerative discs of rat tail before and after controlled traction.METHODSSix-month-old male Sprague-Dawley rats were randomly divided into seven groups: Group A: control group (instrumented with Kirschner [K]-wires only); Group B: Model group (caudal vertebrae immobilized using a custom-made external device to fix four caudal vertebrae [Co7−Co10], while Co8−Co9 vertebrae underwent 4 weeks of compression to induce disc degeneration); Group C: experimental control group (devices removed after the 4 week compression described in Group B, and recovered by themselves for 4 weeks). The remaining four groups represented intervention groups (Groups D and F: Co8−Co9 vertebrae compressed for 4 weeks followed by 2 or 4 weeks of in situ traction, respectively; Groups E and G: vertebrae compressed for 4 weeks followed by 2 or 4 weeks of excessive traction, respectively). X-ray and magnetic resonance imaging were performed at each time point to measure disc height and T2 signal intensity. At the end of the experiment, the animals were euthanized and tail vertebrae harvested for analysis of intervertebral disc histopathology, proteoglycan content, elastic modulus of fibers of the annulus fibrosus (AF) and nucleus pulposus (NP), and microstructure of the bony end plate.RESULTSAfter 2 to 4 weeks of continuous traction (in situ and excessive traction), the Co8–Co9 intervertebral disc space of rats in Groups D to G increased significantly compared with Groups B and C (p < .05). In addition, signs of tissue regeneration were apparent in all four intervention groups (D–G). In addition, histologic scores of the intervention groups (D–G) were significantly lower than those in the model and experimental control groups (Groups B and C, respectively), although no significant difference was found between those four groups. Compared with the model group (Group B), total proteoglycan content of the NP in the intervention groups (D–G) increased significantly (p < .05). After 2 to 4 weeks of intervention (in situ and excessive traction), the morphology of pores in the bony end plate, their number, and the diameter had recovered significantly compared with those in Group B. The in situ traction group was superior to the excessive traction group, and 4 weeks in situ group significantly superior to the 2 weeks group. In all intervention groups, in both the inner and outer AF, mean fibril diameter decreased significantly (p < .05), although they remained larger in the excessive traction group than that in the in situ traction group. Consistent with trend in collagen fiber diameter, the outer AF was stiffer than the inner, and the modulus of the AF in each intervention group not significantly different from that of the control group (Group A) except Group C. However, within the NP, the variation in trend in diameter and modulus of collagen fibers was essentially inconsistent with that of the AF.CONCLUSIONSDegenerated discs exhibit greater reconstruction after low tension traction. It is clear that the intervertebral disc mechanical microenvironment depends to a greater extent on low-tension traction than high-tension traction.     

Nucleus pulposus allograft retards intervertebral disc degeneration

Autogenous implantation of nucleus pulposus or nucleus pulposus cells that were activated by coculture retards intervertebral disc degeneration, but harvesting such grafts causes disc degeneration at the donor site. This study examined whether nucleus pulposus allografts similarly retard disc degeneration and whether such allografting induces immunologic rejection. Japanese White rabbits served as donors and recipients for allografts. Lumbar disc degeneration was induced by aspirating the nucleus pulposus. Two weeks later, intact nucleus pulposus or nucleus pulposus cells were injected and compared with a sham procedure and normal control. The recipients' discs were examined histologically and immunologically at intervals for 16 weeks. Discs receiving an intact nucleus pulposus showed the least degeneration, followed by discs receiving nucleus pulposus cells, both of which were better than no treatment. These findings correlated directly with the intensity of immunochemical staining for Type II collagen. Allogeneic grafts did not induce any appreciable host-versus-graft response. Injection of nucleus pulposus and nucleus pulposus cells retards intervertebral disc degeneration. However, injection of intact nucleus pulposus is more effective than injection of nucleus pulposus cells alone. The intercellular matrix plays an important, but poorly understood, role in preserving intervertebral discs.     

Role of leukocytes in radicular pain secondary to herniated nucleus pulposus

Some studies have assessed inflammatory cells such as macrophages, lymphocytes, and neutrophils in herniated lumbar disc tissues using histologic analysis. However, there is no consensus regarding the relationships between clinical symptoms, including radicular pain and the presence of inflammatory cells. It has been shown that autologous nucleus pulposus relocated on the lumbar nerve root in rats produces time dependent and reversible mechanical hyperalgesia, which is thought to be a pain related behavior in peripheral neuropathic pain models. The purpose of this study was to determine whether leukocytes play a role in the mechanical hyperalgesia induced by the nucleus pulposus and to characterize the role of leukocytes in radicular pain attributable to lumbar disc herniation. Nitrogen mustard was used to induce and evaluate leukocytopenia in rats. Sensitivity to mechanical noxious stimuli was measured quantitatively, and inflammatory cells in granulation tissue around the nerve root were examined histologically. The nucleus pulposus produced neither mechanical hyperalgesia nor abundant inflammatory cells in rats with nitrogen mustard induced leukocytopenia. Neuropathic pain produced by the nucleus pulposus, when placed on the nerve root, may be related to inflammatory cell infiltration induced by relocation of the nucleus pulposus, rather than the nucleus pulposus itself.     

两种骨水泥渗漏对椎间盘影响的实验研究

目的 探讨椎体成形术时骨水泥渗漏是否会引起椎间盘退变，以及椎间盘退变程度与骨水泥类型是否相关。方法 选用8只成年家犬，以每只犬L2-3、L3-4、L4-5椎间盘为实验对象，随机分为对照组、聚甲基丙烯酸甲酯（polymethylmethacrylate，PMMA）与磷酸钙骨水泥（calcium phosphate cement，CPC）3组。对照组仅行椎间盘穿刺，不注入任何物质，PMMA组及CPC组均各向椎间盘注入0．1ml骨水泥。术前及术后24周摄正、侧位X线片，计算椎间盘高度指数百分数（disc height index percentage，DHIP）。术后24周行MR检查，计算MRI指数。组织学检查参照Masuda标准对椎间盘退变程度评分并分析。结果 术后24周X线片显示对照组椎间隙无狭窄，病理学检查未见椎间盘退变。PMMA、CPC组椎间盘MRI显示：椎间隙有狭窄，R加权像髓核信号不同程度降低且不均一，其相对高信号区面积减小，髓核形态不规则，纤维环与髓核界限不清。组织学检查显示髓核细胞数量不同程度减少，空泡变小。髓核的细胞外基质不同程度压缩，纤维环断裂或扭转。3组DHIP、MRI指数、组织学评分的差异均有统计学意义（P〈0.01）。结论 PMMA、CPC注入椎间盘会导致椎间盘退变，PMMA所致椎间盘退变较CPC更为严重.     

Histological changes in intervertebral discs after smoking and cessation: experimental study using a rat passive smoking model

Background Passive smoking has been reported to induce intervertebral disc degeneration in rats, and the objective of the present study was to histologically investigate changes in smoking-induced intervertebral disc degeneration after cessation of smoking. Methods Four-week-old rats were subjected to passive smoking for 8 weeks in a smoking box [20 cigarettes a day: one cigarette an hour (inhaled over 3 minutes and followed by ventilation with room air for 5 minutes)] to induce intervertebral disc degeneration. Smoke-free periods of different lengths were then established, and intervertebral discs were histologically analyzed. Results Immediately after 8 weeks of passive smoking, intervertebral discs exhibited cracks, tears, and misalignment of the annulus fibrosus, and increased fibrous tissue was seen in the nucleus pulposus. In addition, the level of interleukin-1β in intervertebral discs was higher in the smoking group than in the non-smoking group. After cessation, progression of degeneration ceased, and the matrix of the nucleus pulposus and annulus fibrosus exhibited increased fibrous connective tissue and proteoglycan. However, there were no changes in annulus fibrosus misalignment. Interleukin-1β levels also remained significantly elevated after 8 weeks of cessation. Conclusions While the annulus fibrosus degeneration caused by smoking was partially irreversible after cessation of smoking, the amount of mucin (proteoglycan) in the nucleus pulposus and annulus fibrosus tended to increase after cessation, thus suggesting the possibility that smoking-induced intervertebral disc degeneration can be repaired to some degree by cessation of smoking.     

组织工程化同种异体椎间盘移植的动物实验研究

目的观察组织工程技术的运用能否延缓椎间盘移植后的退行性改变。方法将髓中受细胞复合至同种异体椎间盘,体外培养后植入犬L4／k椎间隙作为实验组（A组）,对照组（B组）行同种异体椎间髓移植。使用影像学、生物力学及组织学分析评估植入椎间盘的转归并行组间比较。结果移植椎间盘可与宿主椎体实现骨性融合。对照组椎间盘术后退变明显,12周时其椎间盘高度及髓核信号比灰度值明显低于实验组,稳定性丧失明显;组织学观察发现实验组移植椎间盘结构保持较好,髓核细胞数量较多,排列规则;对照组髓核形态保持欠佳,结构紊乱,髓核细胞数量减少,退行性改变明显。结论通过复合种子细胞实现异体椎间盘的组织工程化可有效延缓椎间盘移植后的退行性改变。     

2000 Volvo Award winner in basic science studies: Exogenous tumor necrosis factor-alpha mimics nucleus pulposus-induced neuropathology. Molecular, histologic, and behavioral comparisons in rats

STUDY DESIGN: This study tested the hypothesis that the 17-kDa form of tumor necrosis factor-alpha is the pathophysiologic agent expressed by herniated nucleus pulposus in vivo that is primarily responsible for the histologic and behavioral manifestations of experimental sciatica associated with herniated lumbar discs. OBJECTIVE: The authors determined the molecular weight and concentration of active tumor necrosis factor-alpha in rat herniated disc and used exogenous tumor necrosis factor-alpha at the same molecular weight to study its neuropathologic effect on rat nerve root and dorsal root ganglion preparations in vivo. SUMMARY OF BACKGROUND DATA: Expressed by herniated nucleus pulposus in culture, tumor necrosis factor-alpha causes neuropathologic injury in nerve roots and neuropathic pain states in which mechanical allodynia is seen in response to peripheral stimuli. METHODS: Western blotting was used to identify the molecular weight of the operative tumor necrosis factor-alpha protein form, and measures of optical density were used for semiquantitative determination of concentration. Plastic-embedded nerve roots and dorsal root ganglion were used for neuropathologic evaluation, and von Frey stimulation was used to quantify mechanical allodynia. RESULTS: The 17-kDa form of tumor necrosis factor-alpha is expressed by herniated nucleus pulposus at a concentration of approximately 0.48 ng per herniated rat lumbar disc. Exogenous tumor necrosis factor-alpha applied in vivo to rat nerve roots produced neuropathologic changes and behavior deficits that mimicked experimental studies with herniated nucleus pulposus applied to nerve roots. CONCLUSIONS: The data reinforce other evidence that tumor necrosis factor-alpha is involved in mechanisms of neuropathic pain.     

Effect of nucleus pulposus on the neural activity of dorsal root ganglion

STUDY DESIGN: This study was designed to investigate, using neurophysiologic techniques in an in vivo rat model, the effect of application of nucleus pulposus to the nerve root on the neural activity of the dorsal root ganglion and the corresponding receptive fields. OBJECTIVES: To assess a further role of the dorsal root ganglion in mechanisms of radicular pain in lumbar disc herniation. SUMMARY OF BACKGROUND DATA: It has been suggested that the epidural application of autologous nucleus pulposus without mechanical compression causes nerve root inflammation and related radicular pain in lumbar disc herniation. Concerning the dorsal root ganglion, its mechanical hypersensitivity and potential for generating ectopic discharges have been reported. However, the effect of autologous nucleus pulposus on the dorsal root ganglion is uncertain. METHODS: In adult Sprague-Dawley rats spontaneous neural activity was recorded from the surgically exposed L5 dorsal root using electrophysiologic techniques, and the mechanosensitivity of L5 dorsal root ganglia and corresponding receptive fields on the hind paw were measured using calibrated nylon filaments. Autologous nucleus pulposus from the tail or fat was implanted at the L5 nerve root. Neural activity was monitored for 6 hours. RESULTS: Spontaneous neural activity in the nucleus pulposus group gradually increased and showed significant differences compared with the fat group from 2.5 to 6 hours after exposure. The mechanosensitivity of the dorsal root ganglia showed significant increases compared with the fat group. CONCLUSIONS: After application of nucleus pulposus to the nerve root, the dorsal root ganglion demonstrated increased excitability and mechanical hypersensitivity. These results suggest that nucleus pulposus causes excitatory changes in the dorsal root ganglion.     

对腰椎间盘突出症传统机械压迫刺激观的质疑与反思

脊神经根的机械性压迫被认为与疼痛和特定节段神经功能障碍有关,然而不是在所有情况下突出髓核都导致相应的临床症状,解除突出的髓核也并不皆意味着腰腿痛的立即消失。该文通过系统的文献研究,认为椎管内突出髓核是否导致相应的临床症状存在着诸多或然因素,并因此提出腰椎间盘突出症多因素动态致压观。