双上肢去势联合椎间盘刺破诱导建立大鼠椎间盘退变模型

背景：寻求一种相对合适的椎间盘退变动物模型,对深入研究椎间盘退变的发病机制有着重要意义。目的：探索一种新的椎间盘退变动物模型,希望为椎间盘退变疾病的基础研究提供新的造模方式选择。方法：将20只雄性SD大鼠随机分为假手术组与模型组。采用双上肢去势联合椎间盘刺破的方式制备椎间盘退变模型,于造模当天及造模后4周、8周进行MRI检查,观察造模前后椎间盘形态变化并评估Pfirrmann分级;造模4,8周后取出大鼠L₄-L₅及L₅-L₆椎间盘,运用苏木精-伊红染色、Masson染色法观察椎间盘病理学变化及免疫组化观察Ⅱ型胶原蛋白表达量。结果与结论：(1)模型组在造模当天仅呈现轻微退变;(2)在造模第4周时与假手术组相比,模型组MRI上T2加权信号降低,在Pfirrmann分级上可见明显差异(P<0.05),在造模第8周时与假手术组相比,模型组MRI上T2加权信号明显降低,与纤维环边界模糊不清,且髓核向后纵韧带膨出,在Pfirrmann分级上可见显著差异(P<0.01);(3)病理染色结果显示,在造模...     

椎间盘退变模型的研究进展

椎间盘退变疾病发病率越来越高,但退变的机制尚不明确.椎间盘退变模型是现今研究椎间盘退变疾病的主要方式,退变模型主要分为体内退变模型和体外退变模型两大类.两种退变模型从不同角度研究椎间盘退变的病理生理过程,为揭示退变机制及预防、治疗椎间盘退变疾病起着重要作用.本文就目前国内外关于两种不同椎间盘退变模型研究的进展作一综述.     

建立剪切应力导致椎间盘退变模型

目的 建立剪切应力导致椎间盘退变的在体动物模型,探索剪切应力与椎间盘退变之间的关系。方法 20只成年日本大耳白兔随机平均分成2组。加载组通过手术在动物L4/5节段植入自行研制的剪切应力加载装置,并加载50 N的剪切力4周;对照组只植入加载装置但不加载剪切力。4周后处死所有动物并对所有相关节段的椎间盘做病理检查。结果 术后的影像学检查显示加载装置位于动物腰椎,手术过程和植入的加载装置对实验动物的日常活动与饮食均无影响。病理学检查发现加载组椎间盘的纤维环组织排列紊乱,正常髓核细胞明显减少。结论 新型装置的设计减少了对动物椎体的创伤,并且可以提供可靠的剪切应力;在体动物模型的建立揭示一定的剪切应力可导致动物椎间盘退变,对进一步研究载荷与椎间盘退变间的关系具有较大的意义。     

低温等离子消融术建立兔椎间盘退变模型的实验研究

目的　探讨低温等离子消融术建立兔椎间盘退变模型的方法与可行性。 方法　新西兰大白兔24只,随机分为实验组12只,对照组12只。两组均穿刺L3/4～L5/6椎间隙,实验组采用消融30s建模,对照组单纯以穿刺针穿刺建模。分别于术前及术后4、8、12周行CR、MRI检查及病理学检查。 结果　实验组DR及MR检查至术后12周均可见退变逐步加重的征象,如椎间隙高度的丢失及T2信号逐步降低。对照组DR及MR检查4周后无明显的退变加重迹象。两组病理学检查均未见早期的髓核缺失,实验组12周可见髓核正常结构消失,椎间盘内结构紊乱。 结论　低温等离子消融术比传统单纯穿刺更易及快速建立椎间盘退变的动物模型,采用此方法建立兔椎间盘退变模型是可靠可行的。     

兔腰椎间盘退变模型的建立及影像学分析

目的：建立腰椎间盘退变的动物模型并进行CR和MRI观察.方法：选用新西兰兔20只,沿右腹直肌外缘做15 cm长切口,钝性剥离腹膜至腰椎横突前外侧,咬除右侧L5、L6横突,显露上述节段椎间盘,斜形切开纤维环约1.5 mm,未伤及髓核,然后逐层缝合.所有动物在标准条件下饲养,分别于术后2、4、8、20、40周行腰椎计算机x线摄影术（CR）和核磁共振成像（MRI）以检测终板下骨及髓核的变化.结果：术后作为自体对照组的L1、2、L2、3椎间盘未见异常,而手术组L4、5、L5、6椎间盘则相继出现T2加权像低信号、腰椎不稳畸形,终板下骨质硬化,椎体边缘骨赘增生,椎间隙变窄,椎间盘后突和硬膜囊受压等改变.对手术节段及其邻近和完全正常节段椎间盘髓核信号值的定量分析显示,手术组椎间盘T2加权像信号值减低在术后4、8、20、40周与正常对照组对比具有统计学意义,而临近椎间盘L3、4、L6、7手术8周后与正常椎间盘对比有显著差异.CR扫描结果显示：手术节段椎间盘终板下信号值减低与对照组相比4周后就开始有显著差异.结论：应用纤维环切开法可获得可靠的新西兰大白兔椎间盘退变模型,可通过MRI及CR在早期加以证实.     

椎间盘退变模型的研究现状及进展

目的：探讨椎间盘退变模型的研究现状及进展。方法：在Pubmed、中国知网查阅有关椎间盘退变模型研究的文献,进行汇总分析。结果：椎间盘退变模型可以通过体外和体内两种方法构建,前者包括椎间盘细胞模型和椎间盘组织模型,后者包括诱发性椎间盘退变模型和自发性椎间盘退变模型。体外模型适用面较广,但培养要求较高,不能全面模拟体内环境;体内培养干预技术较容易实现,但适用面较窄。结论：人椎间盘退变的影响因素繁多,目前的椎间盘退变模型往往具有一定的局限性,无法全面地模拟出人的椎间盘退变情况。无创、微创构建椎间盘退变模型将是未来的发展趋势。     

犬椎间盘经皮针刺退变模型的建立

背景：普通针头穿刺纤维环是目前较为公认的理想椎间退变模型制作方法,但存在对动物损伤大,操作不简捷等缺点。 目的：尝试在C臂机透视下经皮穿刺比格犬腰椎椎间隙建立腰椎间盘退变动物模型。 方法：选用8只健康成年犬,行腰椎MRI后在C臂机下经皮穿刺椎间隙,损伤L5/6椎间盘。术后3,6个月行腰椎MRI后,取标本行免疫组织化学观察椎间盘退变情况。 结果与结论：经皮椎间盘穿刺后3,6个月,L5/6椎间盘均出现了明显退变,MRI显示T2值均出现明显降低,并且椎间盘大多出现不同程度椎间盘突出征象;L5/6穿刺节段髓核Ⅱ型胶原阳性细胞含量较L4/5髓核均明显减少(P < 0.05)。说明经皮穿刺损伤椎间盘建立犬椎间盘退变模型是一种简单有效、重复性好的建模方法。     

纤维环穿刺法与髓核抽吸法建立兔椎间盘退变模型的比较

目的:比较采用纤维环穿刺法和髓核抽吸法建立兔椎间盘退变动物模型的不同。方法:新西兰大白兔经腹膜外入路暴露腰3/4、腰4/5、腰5/6椎间隙,纤维环穿刺组采取针刺纤维环,髓核抽吸组抽吸髓核8周后核磁共振观察椎间盘变化,生化分析椎间盘中蛋白多糖含量,免疫组织化学检测Ⅱ型胶原表达。结果:术后8周与对照组相比,采用两种方法的椎间隙高度与T2信号手术前后变化比值,以及Ⅱ型胶原和蛋白多糖含量,差异显著。其中髓核抽吸组的Ⅱ型胶原和蛋白多糖含量明显低于纤维环穿刺组。结论:髓核抽吸法建立兔退变椎间盘模型退变程度大于纤维环穿刺法。     

纤维环损伤诱导兔椎间盘退变模型

目的:探讨纤维环刺伤法诱导腰椎间盘退变建立动物模型的可行性.方法:普通级新西兰大白兔(雄性,体质量约3.5 kg,1岁龄).分为正常对照组及模型组,以腰4～5椎问盘为正常对照组,以每只兔的腰2～3椎间盘为模型组.用16号穿刺针于腰2～3椎间盘纤维环前外侧刺入,分别于术前及术后4、8、16周对模型组及正常对照组椎问盘进行磁共振(MRI)检查,动物处死取材后行组织学及免疫组织化学观察.结果:术后4～16周,模型组椎间盘髓核MRI T<,2>W<,1>信号呈逐渐减弱趋势,组织学观察髓核脊索细胞逐渐减少,免疫组织化学观察到髓核中Ⅰ型胶原表达较正常对照组逐渐增多,而Ⅱ型胶原表达较对照组逐渐减少.结论:本实验可为椎间盘退变的进一步研究提供动物模型.     

兔椎间盘退变与修复模型的研究现状*

椎间盘退变模型是研究椎间盘退变疾病的基础和关键之一。兔退变椎间盘模型具有操作简单、可重复性好等特点被国内外学者广泛应用。兔椎间盘退变模型包括体内模型、体外模型等。体内模型根据损伤类别包括:机械损伤模型、化学损伤模型、异常应力模型、脊柱不稳模型、脊柱融合模型等;体外模型包括椎间盘细胞模型、椎间盘组织模型等。本文根据近年兔腰椎间盘各种退变模型与修复的研究现状与进展作一综述。     

Experimental model of intervertebral disc degeneration by needle puncture in Wistar rats

Animal models of intervertebral disc degeneration play an important role inclarifying the physiopathological mechanisms and testing novel therapeuticstrategies. The objective of the present study is to describe a simple animalmodel of disc degeneration involving Wistar rats to be used for researchstudies. Disc degeneration was confirmed and classified by radiography, magneticresonance and histological evaluation. Adult male Wistar rats were anesthetizedand submitted to percutaneous disc puncture with a 20-gauge needle on levels 6-7and 8-9 of the coccygeal vertebrae. The needle was inserted into the discsguided by fluoroscopy and its tip was positioned crossing the nucleus pulposusup to the contralateral annulus fibrosus, rotated 360° twice, and held for 30 s.To grade the severity of intervertebral disc degeneration, we measured theintervertebral disc height from radiographic images 7 and 30 days after theinjury, and the signal intensity T2-weighted magnetic resonance imaging.Histological analysis was performed with hematoxylin-eosin and collagen fiberorientation using picrosirius red staining and polarized light microscopy.Imaging and histological score analyses revealed significant disc degenerationboth 7 and 30 days after the lesion, without deaths or systemic complications.Interobserver histological evaluation showed significant agreement. There was asignificant positive correlation between histological score and intervertebraldisc height 7 and 30 days after the lesion. We conclude that the tail discpuncture method using Wistar rats is a simple, cost-effective and reproduciblemodel for inducing disc degeneration.     

新型兔腰椎间盘退变模型的建立

目的：采用兔腰椎间盘内显微注射120KDa Fn片段构建腰椎间盘退变动物模型，并探讨该模型的有效性和优势。方法：新西兰大白兔30只，侧前方入路前方暴露L4-6椎间盘，微量注射器于L4-6注射120kDa Fn片段作为试验组，L，注射PBS作为对照组。分别于第2、4、8、12、16周对椎间盘进行X线、MRI、大体解剖和HE、Massion、高碘酸-Schiff染色组织病理学检测。结果：大体组织观察见术后第8周开始纤维环各层间分界模糊，外侧可见瘢痕样增生，16周出现骨样组织形成。病理染色见术后4周开始纤维环胶原出现断裂等退行性变，蛋白多糖含量降低。MRI观察发现第12周髓核高信号区强度下降，16周信号基本消失。对照组无明显变化。结论：120kDa Fn片段是诱导兔腰椎间盘退变简单、确切的方法，该方法为建立腰椎间盘退变模型提供了新思路，建立了新的腰椎间盘退变动物模型。     

The cell biology of intervertebral disc aging and degeneration

Intervertebral disc degeneration, which mimics disc aging but occurs at an accelerated rate, is considered to be related to neck or low back pain and disc herniation. Degenerated discs show breakdown of the extracellular matrix and thus fail to bear the daily loadings exerted on the spine. Rather than a passive process of wear and tear, disc degeneration is an aberrant, cell-mediated response to progressive structural failure due to aging and other environmental factors such as abnormal mechanical stress. With aging and degeneration, disc cells undergo substantially biologic changes, including alternation of cell type in the nucleus pulposus, increased cell density but decreased number of viable cells as a result of increased cell death and increased cell proliferation, increased cell senescence, and altered cell phenotype which is characterized by compromised capability of synthesizing correct matrix components and by enhanced catabolic metabolism. These changes are involved in the process of disc degeneration through the complicated interactions among them. To retard or reverse disc degeneration, the abnormal conditions of the decreased viable cell population and the altered cell phenotype should be corrected. As potential therapies for disc degeneration, intradiscal protein injection, gene transfer and cell implantation are being understudied in vivo. Suppression of excessive apoptosis and accelerated senescence of disc cells may be other choices for treating disc degeneration. When performing a biologic therapy in order to repair or regenerate the degenerated disc, nutrient and biomechanical factors should also be incorporated, because they are the major causes of the biologic changes experienced by disc cells. Moreover, a very early intervention is indicated by the finding that the onset of human disc degeneration occurs as early as by adolescence.     

The role of nerve fibers and their neurotransmitters in regulating intervertebral disc degeneration

Intervertebral disc degeneration (IVDD) has been the major contributor to chronic lower back pain (LBP). Abnormal apoptosis, senescence, and pyroptosis of IVD cells, extracellular matrix (ECM) degradation, and infiltration of immune cells are the major molecular alternations during IVDD. Changes at tissue level frequently occur at advanced IVD tissue. Ectopic ingrowth of nerves within inner annulus fibrosus (AF) and nucleus pulposus (NP) tissue has been considered as the primary cause for LBP. Innervation at IVD tissue mainly included sensory and sympathetic nerves, and many markers for these two types of nerves have been detected since 1940. In fact, in osteoarthritis (OA), beyond pain transmission, the direct regulation of neuropeptides on functions of chondrocytes have attracted researchers’ great attention recently. Many physical and pathological similarities between joint and IVD have shed us the light on the neurogenic mechanism involved in IVDD. Here, an overview of the advances in the nervous system within IVD tissue will be performed, with a discussion on in the role of nerve fibers and their neurotransmitters in regulating IVDD. We hope this review can attract more research interest to address neuromodulation and IVDD itself, which will enhance our understanding of the contribution of neuromodulation to the structural changes within IVD tissue and inflammatory responses and will help identify novel therapeutic targets and enable the effective treatment of IVDD disease.     

椎间盘再生与修复的组织工程支架材料研究进展

椎间盘退变性疾病是导致下腰痛的主要原因之一,传统保守治疗或手术治疗均存在明显局限性。椎间盘组织工程学的发展为椎间盘退变性疾病的治疗提供了新的治疗方案。支架材料作为构建组织工程椎间盘的关键环节,是目前研究的热点和难点。目前常用的支架材料有天然材料、合成材料及复合材料。本文就构建纤维环、髓核和全椎间盘3个方面支架材料的研究现状进行简要综述。虽然组织工程椎间盘还面临着诸多问题,但未来会为椎间盘退变性疾病的治疗带来美好前景。