磁共振动态增强扫描成年山羊腰椎髓核：定量观察椎间盘的强化程度及营养状态

背景：营养物质的减少是诸多因素导致椎间盘退变的最终途径。许多研究表明椎间盘的营养主要通过终板途径,终板的退变与椎间盘退变也有着密切的关系。MRI动态增强扫描作为一种无创、简单、可重复的方法,逐渐被用于分析椎间盘的营养机制。 目的：观察成年山羊腰椎髓核的MRI动态增强扫描特点。 方法：选取健康成年山羊8只,体质量分别为38 kg及50 kg,共纳入32个腰椎髓核。行腰椎间盘常规矢状位T1-TSE-SPIR、T2WI-TSE-SPAIR和MRI动态增强扫描,观察髓核图像的信号特点,测量增强前及增强后0 min,5 min,10 min,30 min,1 h,1.5 h,2 h,2.5 h,3 h,3.5 h,4 h的髓核信号强度值,分析髓核时间-增强率曲线及峰值时间。 结果与结论：32个髓核在10 min内未见明显的强化,在30 min后开始上升,在3.5 h到达最高值,并处于平台期;4 h内未见明显的下降。提示正常成年山羊腰椎髓核的MRI动态增强扫描曲线具有一定的特征性,观察其MRI动态增强扫描曲线有利于进一步阐明髓核的营养机制。     

针刺损伤兔椎间盘退变模型组织病理变化及软骨样细胞的迁移规律**☆

背景：兔椎间盘退变模型间盘退变表现为随时间进展脊索细胞将被软骨样细胞逐渐替代,但兔针刺纤维环间盘退变模型中软骨样细胞的来源和移行规律尚不明确。 目的：观察针刺兔纤维环间盘退变模型椎间盘病理变化过程,并初步探讨软骨样细胞来源及移行规律。 方法：将24只新西兰大白兔随机分为手术组与假手术组。手术组使用16 G穿刺针针刺L2/L3、L3/L4、L4/L5及L5/L6椎间盘纤维环,假手术组暴露至相同椎间盘前方后冲洗闭合伤口。 结果与结论：针刺损伤椎间盘退变过程中的软骨样细胞来源于终板。在髓核与上下终板交界处,软骨细胞脱离终板成串向髓核中心迁移;在髓核与内层纤维环交界处,软骨细胞沿纤维走行迁移并随之向皱缩的髓核边缘迁移。椎间盘退变过程中非钙化层逐渐变薄,非钙化层/钙化层比值逐渐降低。 关键词：椎间盘退变;软骨细胞;软骨终板;纤维环穿刺;新西兰大白兔  doi:10.3969/j.issn.1673-8225.2012.09.018     

软骨终板形态与椎间盘退变的相关性

背景：以往研究证明多种内环境因素共同作用引发椎间盘退变,最重要的机制为椎间盘软骨终板的退变。 目的：分析椎间盘退变与终板形态的关系。 方法：回顾性分析62例因椎间盘源性慢性下腰痛和79例因髓核脱出致神经根性症状患者的腰椎MRI正中矢状位图像资料。根据腰椎MRI正中矢状位T1W1图像确定终板形态,T2W1图像确定椎间盘退变程度分级。 结果与结论：平坦型和不规则型终板最常见于椎间盘退变人群下腰椎,L5/S1平坦型最多见。髓核脱出组与椎间盘源性慢性下腰痛组中凹陷型终板椎间盘退变程度均较平坦型、不规则型低,平坦型终板椎间盘退变程度较不规则型低(P < 0.01)。两组间凹陷型与不规则型终板椎间盘退变程度差异无显著性意义,髓核脱出组平坦型椎间盘退变程度较椎间盘源性慢性下腰痛组高(P < 0.05)。提示随着椎间盘退变程度的加重,软骨终板形态有由凹陷型向平坦型、不规则型依次转变的趋势。     

Calcium pyrophosphate dihydrate deposition in the intervertebral discs in a case of Wilson''s disease

The vertebral column from a known case of Wilson's disease (hepatolenticular degeneration) was examined by radiological, histological, histochemical and x-ray microanalytical techniques which demonstrated the presence of focal depositions of calcium pyrophosphate dihydrate (CPPD) in the intervertebral discs. These deposits were present in both the annulus fibrosus and the nucleus pulposus but in certain discs the deposits were concentrated near the interface between disc and vertebral body bone endplates. At these sites there was new bone formation with narrowing of the discs, irregularity and sclerosis of the bone endplates and exostosis.     

Proteoglycans and collagen in the intervertebral disc of the rhesus monkey (Macaca mulatta).

The different varieties of supportive tissues in the intervertebral discs of the rhesus monkeys (Macaca mulatta) were investigated with regard to morphology of the collagen fibrils and distribution and localization of proteoglycans (PG). The annulus fibrosus and the nucleus pulposus of the intervertebral disc were structurally closely integrated into the cartilaginous endplates of the vertebral bodies. The collagen fibrils in the intervertebral disc fell into two categories; i.e. thick (70-110 nm) and thin (40-50 nm) ones. In the outer zone of the annulus fibrosus only thick fibrils occurred, while in the regions of the inner part of the annulus fibrosus, in the periphery of the nucleus pulposus and in the cartilaginous endplates both types of fibrils were found. PG were found free in the matrix and in great numbers in association with collagen fibrils in all regions of the intervertebral disc. They interconnected neighbouring fibrils and decorated the surface of the collagen fibrils in irregular orientation. Only the thick fibrils of the annulus fibrosus contained small regularly arranged intrafibrillar PG precipitates. The free precipitates were usually longer and thicker than the collagen-associated PG, in addition they could be branched. Since rhesus monkeys are closely related to humans, they can serve as model organisms and the findings presented may be of relevance to the understanding of the human intervertebral discs.     

骨形态发生蛋白2对退变椎间盘组织学和Ⅰ，Ⅱ型胶原的影响

背景：研究表明,退变椎间盘细胞外基质的主要变化是Ⅱ型胶原和蛋白多糖含量的减少和Ⅰ型胶原含量的增加。 目的：观察腺相关病毒介导的骨形态发生蛋白2基因(AAV-BMP-2)对兔退变腰椎间盘内髓核Ⅰ,Ⅱ型胶原的影响。 方法：将12只新西兰大白兔L2-3,L3-4,L4-5,L5-6椎间盘针刺制造退变模型后随机分为3组,每组4只。其中AAV-BMP-2组兔椎间盘注射AAV-BMP-2,AAV组兔椎间盘注射不携带目的基因的空载体,生理盐水组兔椎间盘注射生理盐水,注射后第8周处死动物取其相应的椎间盘常规石蜡包埋,组织切片,以苏木精-伊红染色观察其髓核组织学变化,免疫组织化学法检测髓核组织Ⅰ型、Ⅱ型胶原表达并进行半定量分析。 结果与结论：普通苏木精-伊红染色结果显示AAV-BMP-2组椎间盘内髓核细胞数目较多,呈单个或者簇状分布,髓核结构清晰,无纤维样组织填充。而AAV组和生理盐水组的组织结构相似,髓核内细胞数目少,髓核皱缩干瘪,细胞间为纤维样组织填充且排列紊乱。免疫组织化学染色显示：AAV-BMP-2组椎间盘髓核内Ⅱ型胶原的表达均高于AAV组和生理盐水组(P < 0.05);AAV-BMP-2组椎间盘髓核内Ⅰ型胶原的表达均低于AAV组和生理盐水组(P < 0.05)。结果说明,体内转染AAV-BMP-2能抑制椎间盘髓核细胞表达Ⅰ型胶原,促进椎间盘髓核细胞表达合成Ⅱ型胶原,提示维持椎间盘内胶原的含量、种类,可维持椎间盘内组织学的结构和形态,稳定髓核细胞生长环境,延缓椎间盘的退变。     

椎间盘突出患者椎间盘Fas基因的表达

背景：临床中发现腰椎间盘突出症常在一个家族中多人甚至全部发病,而且发病的部位、原因、症状基本一致,这表明基因在该种病症中扮演着重要的角色。 目的：分析家族性腰椎椎间盘突出症患者椎间盘内Fas凋亡基因表达的特点。 方法：用半定量RT-PCR方法检测15例家族性腰椎椎间盘突出患者、21例普通椎间盘突出患者、5例新鲜尸体椎间盘的软骨终板和髓核组织中Fas基因的表达情况。 结果与结论：家族性椎间盘突出与普通椎间盘突出患者终板内Fas基因的表达均高于新鲜尸体椎间盘终板内Fas基因的表达,差异有显著性意义(P < 0.05),家族性椎间盘突出与普通椎间盘突出患者终板内Fas基因的表达差异无显著性意义(P > 0.05)。家族性椎间盘突出患者髓核内Fas基因表达与普通椎间盘突出患者、新鲜尸体髓核内Fas基因表达相比较,差异无显著性意义(P > 0.05)。结果可见家族性椎间盘终板中Fas基因的表达增加可能是继发性的,可以通过阻止终板退变来达到阻止椎间盘退变的目的。     

成人腰椎终板冠状面后缘解剖形态学分析及其临床意义

目的　观测正常成人腰椎终板冠状面后缘的CT解剖学特征，为临床椎间融合器设计及腰椎间盘置换术等手术提供解剖学依据。 方法　将62套成人腰椎L1~5上、下缘终板冠状面后缘形态进行解剖学分型，分为3型：I型，凹形；Ⅱ型，平型；Ⅲ型，凸型，测量（1）上、下终板左侧椎弓根与终板连接的中点分别到终板中线的垂线距离（LLU，LLL）；（2）上、下终板右侧椎弓根与终板连接的中点分别到终板中线的垂线距离（LRU，LRL）；（3）上、下终板冠状面后缘中点分别到终板中线之间的垂线距离（LMU，LML）。 结果　L1~4上、下终板形态以凹型为主，L1上终板凹型占93.75%，下终板占80%；L2上终板凹型占77.27%，下终板占68.18%；L3上终板凹型占58.33%，下终板占54.17%；L4上终板凹型占66.67%，下终板占55.56%。L5上终板则以凹型和凸型为主要类型，分别为35.48%、35.48%；下终板则以凸型为主要类型，占61.29%。在L2上终板，LLU、LRU、LMU存在性别间的差异，有统计学意义（P<0.05）；在L3下终板，LLL、LRL存在性别间的差异，有统计学意义（P<0.05）。 结论　L1~4上、下终板的主要类型为凹型，而L5上、下终板较为特殊，上终板主要为凹型和凸型，下终板多为凸型。本研究对腰椎上、下终板形态分型及测量可作为人工腰椎间盘终板的设计及改进的参考依据。     

脊柱内固定后椎间盘营养途径的实验研究

目的：为了研究脊柱内固定后椎间盘的营养途径。方法：24只日本大耳白兔，行墨汁灌注后，观察同一动物非固定区中椎间盘与固定区域椎间盘的墨汁分布情况，观察相同位置对照组与实验组的墨汁分布情况。结果：软骨终板有血管从中经过，椎间盘营养来源分主要和次要途径，内固定后软骨终板和椎体之间产生分离，椎间盘会新生较多小血管，且血管缓慢而淤积。结论：内固定节段椎间盘处于低应力状态下，一定时间后，营养代谢的改变会导致椎间盘退行样变的发生。     

等离子低温消融对椎间盘髓核形态学的影响

目的探讨等离子低温髓核消融术中不同的消融时段椎间盘髓核组织的形态学改变。方法选择2月龄实验用小型猪6只,平均体重22.5kg,雌雄不计,选取L1～S1共30个椎间盘,随机分为5组消融,每组消融时间分别定为0、30、60、90和120s。术后按照设定的时间取出完整椎间盘,取出髓核及终板行HE染色,在普通光学显微镜下观察病理变化。结果消融后髓核组织主要表现为基质成分减少,髓核细胞聚集。随消融时间延长,消融区域髓核中心出现组织空洞,消融区域周围出现染色均匀的条带样基质。继续增加消融时间至120s,消融区域无扩大,但区域内软骨细胞数量减少,部分软骨终板完整性受破坏。结论等离子低温消融可使髓核内基质成分减少、细胞聚集,但延长消融时间并不能扩大消融范围,且可对软骨终板造成损伤。     

腰椎退行性变终板形态影像学表现及临床意义

目的 探讨下腰痛患者的软骨终板形态改变,并进一步研究软骨终板形态与腰椎间盘退变的关系。 方法 回顾性研究84例下腰痛患者。所有患者均行腰椎普通X线及MRI检查,依据MRI正中位像进行软骨终板形态分类及T2像对腰椎间盘退变分级,在侧位平片上进行椎间盘高度测量。 结果 退变腰椎软骨终板形态以扁平型多见。腰椎退行性变其椎间盘退变程度随凹形、扁平形到不规则形逐渐加重,且各型相比有显著性差异;退变椎间盘高度随凹形、扁平形到不规则形逐渐降低,但扁平形与不规则形相比统计学差异不明显。 结论 腰椎退行性变中扁平型软骨终板患者出现几率较高。凹形、扁平型和不规则形腰椎退行性变患者的腰椎间盘退变程度逐渐加重,扁平型和不规则形椎间隙高度降低明显,这对椎间盘髓核假体置换术具有一定指导意义。     

正常成人腰椎间盘相关参数的测量和意义

目的　测量正常成人腰椎间盘相关参数,为设计符合国人腰椎结构特征的人工腰椎间盘和人工髓核提供参数值,同时提供正常人腰椎间隙高度数据,为临床诊断腰椎疾病提供依据。 方法　随机抽取自2010.1~2013.1间,腰背痛患者的MRI阴性图像资料300例（男136例,女164例）。分别测量L1/2~L5/S1 5个节段椎间隙的前高、中高、后高、角度以及L2/3~L5/S1 4个节段的椎间盘矢径、横径和髓核矢径、横径。 结果　各测量值年龄差异不显著（P>0.05）。性别差异显著,且测量值多为男性大于女性。节段差异显著（P<0.05）,椎间盘前高和角度的测量值随着节段增加而递增,中高、后高、椎间盘矢径和横径、髓核矢径和横径的测量值除L5/S1节段外都随着节段的增高而递增。各节段椎间盘的矢径和横径,髓核的矢径和横径间存在线性关系。 结论 腰椎间隙高度测量对腰椎疾病的诊断具有参考价值;在设计人工腰椎间盘和人工髓核时应考虑性别和节段差异,并且应符合矢径和横径间的线性规律。     

In situ oxygen utilization in the rat intervertebral disc

Nucleus pulposus cells of the intervertebral disc have no endogenous vasculature and have thus been hypothesized to be hypoxic. This hypothesis was tested using 2-nitroimidazole, EF5, a drug that at low oxygen concentrations forms covalent adducts with cellular proteins. After administrating EF5 to rats, sections of the intervertebral disc were analysed for EF5 adducts. Drug adducts were quantified in tissue sections using a fluorescent monoclonal antibody. Although the level of EF5 fluorescence in all intervertebral disc tissues was low, the transition zone at the periphery of the nucleus pulposus exhibited the highest level of EF5 binding. To substantiate this result, tissue nitroreductase levels and drug pharmacology were evaluated. Nitroreductase levels were measured in whole discs under severe hypoxia. We noted that there was robust EF5 binding to cells in the annulus fibrosus and transition zone with modest binding to cells of the nucleus pulposus and endplate. High-performance liquid chromatography analysis indicated limitations in EF5 access to the nucleus pulposus, most probably related to the lack of vasculature and slow drug distribution through the gel-like interior of the disc. However, despite diffusion problems, the drug dose was determined to be sufficient to report the oxygen status of the nucleus pulposus cells. Based on these findings, we conclude that despite poor vascularization, the disc cells accommodate to the local environment by displaying a limited need for oxygen. Accordingly, the cells of the intervertebral disc are not severely hypoxic.     

神经肽Y/降钙素基因相关肽在腰椎间盘中的分布与共表达

背景：研究发现,神经肽Y与降钙素基因相关肽在交感神经节中有共存现象。 目的：观察神经肽Y/降钙素基因相关肽在正常腰椎间盘的共分布及在突出腰椎间盘髓核组织中的共表达。 方法：从10例尸体中收集完整的腰椎间盘,在另10例尸体中收集腰椎间盘髓核组织作为对照组。收集30例有症状的腰椎间盘突出症患者,其中L4/5或L5/S1需行腰椎间盘髓核摘除,取出髓核组织作为实验组。 结果与结论：①正常椎间盘组织中神经肽Y/降钙素基因相关肽荧光双标染色阳性的神经纤维较多的分布于椎间盘纤维环外1/3,但在椎间盘内2/3及髓核中未见或少量分布。②髓核的共表达,神经肽Y/降钙素基因相关肽荧光双标染色神经纤维的阳性率实验组均高于对照组(P < 0.05)。提示在正常腰椎间盘组织中神经肽Y/降钙素基因相关肽分布于纤维环外1/3部分,在纤维环内2/3部分及髓核组织未分布,但在突出椎间盘髓核组织中有大量的神经肽Y/降钙素基因相关肽共表达。     

椎间注射转化生长因子β1对兔退变腰椎间盘蛋白多糖表达的影响

背景：体外研究证实转化生长因子β1可以促进蛋白多糖合成,延缓椎间盘退变,但体内实验鲜见报道。 目的：观察局部应用转化生长因子β1对兔退变腰椎间盘髓核蛋白多糖表达的影响。 方法：取30只新西兰大白兔建立兔腰椎间盘退变模型,造模12周,经X射线证实退变后,随机选择6只模型兔及6只未造模正常兔,处死取材。分别向剩余24只模型兔L_4~5椎间隙注射转化生子因子β1和生理盐水。末次给药2,4周取材,间苯三酚法测定兔椎间盘髓核内蛋白多糖的水平。 结果与结论：造模12周,兔退变椎间盘髓核中蛋白多糖水平明显降低(P < 0.01)。经局部注射转化生子因子β1后,兔退变椎间盘髓核中蛋白多糖表达明显增多(P < 0.01)。说明在兔椎间注射转化生子因子β1 可以促进髓核蛋白多糖的合成,延缓椎间盘退变。     

人颈椎间盘退变与细胞凋亡及基质金属蛋白酶11的表达

背景：前期研究发现基质金属蛋白酶11基因在人退变颈、腰椎间盘组织中明显上调。 目的：观察人退变颈椎间盘髓核组织中基质金属蛋白酶11的表达与细胞凋亡的关系。 方法：纳入30个经MRI确认的退变颈椎间盘髓核组织和20个因颈椎创伤治疗获得的正常颈椎间盘髓核组织。 结果与结论：苏木精-伊红染色显示退变的颈椎间盘髓核组织中髓核细胞较正常髓核组织明显减少(P < 0.01),而凋亡细胞较正常髓核组织明显增多(P < 0.01)。免疫组化染色显示退变的颈椎间盘髓核组织中基质金属蛋白酶11的表达明显高于正常髓核组织(P < 0.01),且基质金属蛋白酶11表达与TUNEL染色检测到的细胞凋亡正相关(r=0.44,P < 0.05)。说明高表达的基质金属蛋白酶11不仅可直接破坏细胞外基质尚可诱导髓核细胞凋亡,在椎间盘退变的过程中发挥重要作用。     

Avian intervertebral disc arises from rostral sclerotome and lacks a nucleus pulposus: implications for evolution of the vertebrate disc

Deterioration of the intervertebral discs is an unfortunate consequence of aging. The intervertebral disc in mammals is composed of three parts: a jelly-like center called the nucleus pulposus, the cartilaginous annulus fibrosus, and anterior and posterior endplates that attach the discs to vertebrae. To understand the origin of the disc, we have investigated the intervertebral region of chickens. Surprisingly, our comparison of mouse and chicken discs revealed that chicken discs lack nuclei pulposi. In addition, the notochord, which in mice forms nuclei pulposi, was found to persist as a rod-like structure and express Shh throughout chicken embryogenesis. Our fate mapping data indicate that cells originating from the rostral half of each somite are responsible for forming the avian disc while cells in the caudal region of each somite form vertebrae. A histological analysis of mammalian and nonmammalian organisms suggests that nuclei pulposi are only present in mammals.     

Degeneration alters structure-function relationships at multiple length-scales and across interfaces in human intervertebral discs

Intervertebral disc (IVD) degeneration and associated back pain place a significant burden on the population. IVD degeneration is a progressive cascade of cellular, compositional, and structural changes, which results in a loss of disc height, disorganization of extracellular matrix architecture, tears in the annulus fibrosus which may involve herniation of the nucleus pulposus, and remodeling of the bony and cartilaginous endplates (CEP). These changes to the IVD often occur concomitantly, across the entire motion segment from the disc subcomponents to the CEP and vertebral bone, making it difficult to determine the causal initiating factor of degeneration. Furthermore, assessments of the subcomponents of the IVD have been largely qualitative, with most studies focusing on a single attribute, rather than multiple adjacent IVD substructures. The objective of this study was to perform a multiscale and multimodal analysis of human lumbar motion segments across various length scales and degrees of degeneration. We performed multiple assays on every sample and identified several correlations between structural and functional measurements of disc subcomponents. Our results demonstrate that with increasing Pfirrmann grade there is a reduction in disc height and nucleus pulposus T2 relaxation time, in addition to alterations in motion segment macromechanical function, disc matrix composition and cellular morphology. At the cartilage endplate-vertebral bone interface, substantial remodeling was observed coinciding with alterations in micromechanical properties. Finally, we report significant relationships between vertebral bone and nucleus pulposus metrics, as well as between micromechanical properties of the endplate and whole motion segment biomechanical parameters, indicating the importance of studying IVD degeneration as a whole organ.     

X射线引导建立大鼠椎间盘退变模型

背景:目前国内外对于椎间盘退变的造模方法较多,但存在定位不精准问题.目的:对比X射线引导下与非X射线引导下建立大鼠椎间盘退变模型的效果.方法:取9只成年SD大鼠,按照随机数字表法分3组,每组3只,空白组不进行任何操作;对照组手动定位尾椎Co6/7椎间盘,用注射器针头进行垂直穿刺,并均匀缓慢注入无水乙醇,建立椎间盘退变模...     

山羊腰椎间盘退变模型硬膜外胶原酶的注射

背景：采用椎间盘外、硬膜外腔注射胶原酶治疗椎间盘突出症的机制,目前尚不完全清楚。 目的：观察硬膜外注射胶原酶对椎间盘退变动物模型的作用。 方法：成年山羊6只,麻醉后作侧外方切口至腰椎体腹侧,椎板加压并用钢板螺钉固定,L₁/L₂,L₃/L₄椎间盘分别注射0.5 mL无水乙醇建立椎间盘退变模型。硬膜外注射实验用胶原酶1 mL,2周后处死动物,取出椎间盘,作电镜切片观察。 结果与结论：电镜下显示：退变的椎间盘纤维环上有裂隙,退变的椎间盘的胶原纤维明显溶解,未退变的椎间盘未有明显溶解。提示硬膜外注射胶原酶通过纤维环上的裂隙渗透到盘内,发生化学溶解作用,从而达到治疗作用。