Gene expression profile of degenerated cervical intervertebral disc tissues in rats

egenerationofintervertebraldisciscausedbymanyfactorsandisacomplicatedwebsystemofdifferentialexpressionofmanygenes .Throughstudyingthegeneexpressionprofile ,theanalysisresultsofaseriesofphysiologicalstatesofintervertebraldisccellsortissuescanbeobtainedsensitivelyandcompletely .1,2 Thesedataaremuchvaluabletotheearlydiagnosis ,thecharacteristicpreventionandtherealizationandpopularizationoftherapeuticmethodsforthisdisease ,aswellasprovideprerequisiteandimportantbasisforstudyingthetherapeuticdrugsan…     

椎间盘镜治疗中央型腰间盘突出症附38例临床报告

腰椎间盘突出症是骨科常见病、多发病,其发病率日趋增高,治疗中央型间盘突出以手术为主,传统术式是全椎板切除,经硬膜囊内(或牵开硬膜囊)摘除突出变性的间盘组织,手术破坏了脊柱后柱,增加了脊柱的不稳定性,以致后期腰椎不稳,再次出现腰腿疼症状.随着微创技术引入脊柱外科,椎间盘镜下髓核摘除术的应用避免了后期脊柱不稳的发生,我院2001年初开展此技术,治疗中央型间盘突出38例,取得满意效果,初步报告如下.……     

软骨调节素在成人退变椎间盘中的表达

目的 利用分子生物学及免疫组化方法研究软骨调节素(ChM-Ⅰ)在成人退变椎间盘细胞及椎间盘组织中的表达情况.方法 2009年3月至4月取3例因腰椎间盘退变性疾病而需行后路椎间融合患者的椎间盘组织分别进行髓核及纤维环细胞培养.取部分原代细胞利用RT-PCR和Western blot研究ChM-Ⅰ mRNA和蛋白在成人退变椎间盘细胞中的表达情况.利用real-time PCR和Western blot研究不同浓度碱性成纤维细胞生长因子(bFGF)对髓核细胞和纤维环细胞ChM-Ⅰ mRNA和蛋白表达的影响.收集2008年10月至2009年10月因腰椎间盘退变性疾病而行手术切除的椎间盘组织标本26例,根据MRI表现退变程度为Ⅲ～V级,作为退变组.收集同期因脊柱肿瘤行手术治疗时切除的椎间盘标本6例,退变程度Ⅰ级,作为对照组.利用免疫组化方法研究ChM-Ⅰ在不同退变程度椎间盘组织中的表达情况.结果 RT-PCR、Western blot显示ChM-Ⅰ在椎间盘髓核细胞及纤维环细胞中均有表达.bFGF可抑制ChM-Ⅰ在髓核及纤维环细胞中的表达,且呈剂量依赖性(P＜0.05).ChM-Ⅰ在对照组椎间盘组织中表达量很低,阳性细胞率为0.12±0.03,而在椎间盘发生退变后其表达量明显升高,退变组与对照组相比差异有统计学意义(P＜0.05).结论 髓核细胞及纤维环细胞可表达ChM-Ⅰ,bFGF可明显抑制ChM-Ⅰ mRNA及蛋白的表达.椎间盘发生退变后ChM-Ⅰ表达明显升高,提示其可能在椎间盘退变的病理过程中发挥一定的作用.

Abstract：

Objectives To investigate the expression of chondromodulin-1(ChM-Ⅰ)in human adult degenerative intervertebral disc(IVD)cells and the relationship between ChM-Ⅰ expression and disc degeneration.Methods Three degenerated disc specimens obtained from patients in the treatment of disc degenerative disease from March to April 2009 were used for cell culture.ChM-Ⅰ expression in ⅣD cells was examined by RT-PCR and Western blot.The effect of basic fibroblast growth factor(bFGF)on the expression of ChM-Ⅰ was assessed by real-time PCR and Western blot.From October 2008 to October 2009,26 human ⅣD tissues were obtained from patients in the surgical treatment of disc degenerative disease at different stage of degeneration according to MRI.Six IVD tissues removed from patients with metastatic spinal tumor were used as normal control.The expression of ChM-Ⅰ determined by immunohistochemical analysis was correlated with MRI degeneration grade.Results RT-PCR and Western blot examination showed that ChM-Ⅰ was expressed in both adult degenerative anulus fibrosus and nucleus pulposus cells.The mRNA and protein expression of ChM-Ⅰ were both down-regulated by administration of bFGF with dose-dependent way(P＜0.05).Immunohistochemical analysis showed the percent of ChM-Ⅰ immunopositive cells in the control group was 0.12 ± 0.03,and the number increased significantly in the advanced degeneration group(P＜ 0.05).Conclusions The current results demonstrate that ⅣD cells express ChM-Ⅰ.Administration of bFGF down-regulates the expression of ChM- Ⅰ.The expression of ChM-Ⅰ is correlated with the degree of ⅣD degeneration which means it may involve in the process of ⅣD degeneration.     

Transforming growth factor β receptor induction in herniated intervertebral disc tissue: an immunohistochemical study

Transforming growth factor beta (TGF-beta) is a potent inducer of angiogenesis and fibrogenesis. There is presently little information about the pathophysiological function of TGF-beta in herniated disc tissue. In order to analyze the cellular role and activation of TGF-beta after disc herniation we immunostained frozen material from 38 disc herniation operations and from eight macroscopically normal discs from organ donors. Polyclonal TGF-beta-I, TGF-beta-II and TGF-beta receptor type II antibodies were used with the avidin biotin complex (ABC-) immunoperoxidase method. All the herniated discs were TGF-beta immunopositive. Such immunoreactivity was mainly associated with disc cells. In a few samples, capillaries were also TGF-beta immunopositive. Immunopositivity was similarly observed in the control discs. To analyze possible differences between the two groups, we calculated the ratio of immunopositive disc cells. For all three antibodies, a statistically significantly (Mann-Whitney test, P = 0.0001) higher number of disc cells showed immunopositivity in the herniated discs. The increase in TGF-beta receptor immunopositivity suggested induction of TGF-beta receptors in herniated discs. Our results support an active regulatory role for TGF-beta in disc cell metabolism.     

Osseointegration by bone morphogenetic protein-2 and transforming growth factor beta2 coated titanium implants in femora of New Zealand white rabbits

Background:

Intramembranous bone formation is essential in uncemented joint replacement to provide a mechanical anchorage of the implant. Since the discovery of bone morphogenic proteins (BMPs) by Urist in 1965, many studies have been conducted to show the influence of growth factors on implant ingrowth. In this study, the influence of bone morphogenetic protein-2 (rhBMP-2) and transforming growth factor β2 (TGF-β2) on implant osseointegration was investigated.

Materials and Methods:

Thirty-two titanium cylinders were implanted into the femoral condyles of both hind legs of New Zealand White Rabbits. Four experimental groups were investigated: controls without coating, a macromolecular copolymer + covalently bound BMP-2, adsorbed BMP-2, and absorbed BMP-2+TGF-β2. All samples were analyzed by ex vivo high-resolution micro-computed-tomography after 28 days of healing. Bone volume per total volume (BV/TV) was recorded around each implant. Afterward, all samples were biomechanically tested in a pull-out setup.

Results:

The highest BV/TV ratio was seen in the BMP-2 group, followed by the BMP-2+TGF-β2 group in high-resolution micro-computed-tomography. These groups were significantly different compared to the control group (P < 0.05). Copolymer+BMP-2 showed no significant difference in comparison to controls. In the pull-out setup, all groups showed higher fixation strength compared to the control group; these differences were not significant.

Conclusions:

No differences between BMP-2 alone and a combination of BMP-2+TGF-β2 could be seen in the present study. However, the results of this study confirm the results of other studies that a coating with growth factors is able to enhance bone implant ingrowth. This may be of importance in defect situations during revision surgery to support the implant ingrowth and implant anchorage.     

An individualized and bioactive artificial intervertebral disc

Nowadays, prostheses used for total disc replacement are mostly made of non‐bioactive materials and always predetermined to certain sizes by the manufacturers, resulting in irreversible attrition and non‐individualization. Here, the design of a novel, individualized and bioactive artificial intervertebral disc which avoids the above‐mentioned problems is described, and an evaluation made of it.     

退变腰椎间盘组织中碱性成纤维细胞生长因子的表达研究

目的 探讨碱性成纤维细胞生长因子（basic fibroblast growth factor,bFGF)在正常和退变椎间盘组织中的表达情况。方法 交30例来源于腰椎间盘突出症患者手术中所取得的椎间盘组织（观察组，男11例，女19例；年龄25－78岁，平均48岁；病程3个月－30年，平均9年11个月）与6例来源于脊柱侧凸患者前路松解术所取得的椎间盘组织（对照组，男女各3例，年龄10－17岁，平均14.2岁）进行对比，首先经病理组织学检查证实为退变椎间盘组织和政党椎间盘组织，然后将两组椎间盘组织分别通过免疫组织化学方法和原位杂交方法，检测各自椎间盘组织中的bFGF及其mRNA的表达。观察组30例均为退变椎间盘组织，免疫组化阳性率为90％（27／30），原位杂交阳性率为20％（6／30）；对照组6例均为正常椎间盘组织，其免疫组及原位杂交均为阴性，两组间免疫组化方法检测阳性率在统计学上差异有非常显著性意义。结论 bFGF在正常和退变椎间盘组织中表达的差异有显著性意义。提示 bFGF可能作为增生刺激因子促进椎间盘组织中的软骨细胞增生和细胞外基质合成，进而加速椎间盘退变。     

胶原酶髓核溶解术治疗腰椎间盘突出症概述

胶原酶髓核溶解术是治疗腰椎间盘突出症的一种有限手术,将胶原酶注入突出的椎间盘中或突出髓核周围,通过该酶催化降解髓核的胶原成分,降低髓核内的压力,使突出的椎间盘缩小或回复,减轻或解除对神经根的压迫,达到缓解或消除症状的目的.该方法是介于保守治疗与外科手术之间的方法,1968年哈佛大学医学院Sussman[1]首先提出用胶原酶替代木瓜凝乳蛋白酶溶解突出椎间盘,随后有更多的学者致力于该方法的研究.本文对胶原酶髓核溶解术治疗腰椎间盘突出症的实验与临床研究进展综述如下.……     

退变腰椎间盘髓核组织中NDRG2表达的变化及其意义

[目的]研究退变的腰椎间盘髓核组织中NDRG2表达的变化及其在椎间盘退变中的作用。[方法]收集腰椎间盘标本47例,根据Pfirrmann分级分为Ⅰ~Ⅴ级。采用免疫组织化学染色、实时-定量PCR(RT-PCR)和Western-Blotting技术从细胞、蛋白和基因水平分别检测腰椎间盘髓核组织中NDRG2的表达,并与Pfirrmann分级进行相关性分析;通过RT-PCR分析P53 mRNA的表达;采用衰老相关的β-半乳糖苷酶(SA-β-gal)实验检测腰椎间盘髓核组织中衰老髓核细胞比例,并与NDRG2表达量进行相关性分析。[结果]免疫组织化学染色、RT-PCR及Western-Blotting检测示NDRG2表达随椎间盘退变程度加重而增加,且与Pfirrmann分级呈正相关;P53 mRNA在腰椎间盘髓核组织中的相对表达随着椎间盘退变程度加重而增加,且与NDRG2的表达量呈正相关。SA-β-gal阳性细胞比例在椎间盘髓核组织中随退变程度加重而增加,并且与NDRG2阳性细胞比例呈正相关。[结论]NDRG2参与了腰椎间盘退变的病理过程,并且可能通过介导腰椎间盘髓核细胞衰老对椎间盘退变起促进作用。     

椎体终板下微循环障碍直接导致椎间盘退变的实验研究

目的 通过建立椎体终板下微循环障碍动物模型,探讨椎间盘退变发病的可能机制.方法 将24只新西兰白兔随机分为实验组和对照组,实验组采用联合应用内毒素与激素的方法制备典型椎体终板下微循环障碍模型,并通过终板微血栓染色证实;对照组为阴性空白对照,不给予任何药物干扰,仅标准饲料喂养.3个月后分析实验组和对照组动物椎间盘的水含量、生化成分含量和组织形态学,从而评估椎间盘的退变程度.结果 终板微血栓染色证实实验组成功构建椎体终板下微循环障碍模型,3个月后实验组动物椎间盘水含量、生物化学成分含量均低于对照组,椎间盘切片染色可见椎间盘退变的表现.结论 椎体终板下微循环障碍可直接导致椎间盘退变,椎间盘营养供给障碍是椎间盘退变的发病机制之一.     

儿童颈椎间盘钙化的诊断和治疗

儿童颈椎间盘钙化症（cervical incervertebral dise calcification in children）是一种少见的疾病。1924年Baron首先报道本病．国内1982年至今陆续报道，迄今仅百余例。作者经治儿童颈椎间盘钙化症5例．结合文献对其诊断和治疗加以讨论。     

小切口手术治疗老年性腰椎间盘突出症

我科自1992~2000年6月应用小切口手术治疗老年性腰椎间盘突出症38例,经过6个月~2年的随访,疗效满意。     

生物相容高分子水凝胶在椎间盘修复再生方面作用的研究进展

椎间盘退行性变为慢性疾病,会导致脊柱承重不稳,进而损伤脊髓、马尾神经和神经根,60%的70岁以上老年人群会发病^[1-2]。由于椎间盘退行性变的病理学、病理生理学及生物力学机制并不完全清楚,目前的治疗手段仅能减轻疼痛症状,不能完全消除疾病^[3-4]。椎间盘切除术和椎间融合术是治疗椎间盘退行性变的常用术式,但易造成脊柱生物力学的改变^[5]。人工椎间盘可应用于椎间盘退行性变的治疗,但因椎间盘的结构和功能复杂,很难设计出理想的椎间盘假体来保持天然组织结构和生物力学特征。理想的生物组织替代物应具有高强度、高柔性和高韧性^[6]。     

凝固椎间盘髓核的生物力学研究

目的观察凝固髓核的轴向抗压缩能力,探讨其临床应用的可行性。方法将12只杂种犬的16个椎间盘分为4组：空白对照组4个,明矾溶液1组（注入明矾溶液3d）4个,明矾溶液2组（注入明矾溶液2周）4个,明矾溶液3组（注入明矾溶液1个月）4个。外接压力泵,并维持160kPa压力5min,将0．15ml10％明矾溶液缓慢注入椎间盘髓核,使椎间盘髓核凝固以实验椎间盘为中心,取一个脊柱功能单位作为生物力学检测的标本。检测标本的轴向载荷位移曲线及使标本髓核突出的轴向载荷。结果轴向载荷500N时,明矾溶液凝固的髓核（3个时间组）与正常椎间盘被压缩的位移无统计学差异;轴向载荷2058N（210kg）时,正常椎间盘髓核可被挤出;轴向载荷2538N（259kg）时,明矾溶液凝固的髓核（1个月）被挤出。结论明矾溶液可使椎间盘髓核产生凝固,凝固髓核的轴向抗压能力略有增强,可达到强化髓核的目的。     

微切口潜行开窗术治疗腰椎间盘突出症200例

我院自1997-2001年通过改良手术器械,采用微切口潜行开窗手术治疗腰椎间盘突出症200例,取得满意疗效,现报告如下.……     

破裂型腰椎间盘突出症的临床特征分析

腰椎间盘由退变向纤维环部分破裂,全层破裂,髓核脱出甚至游离至椎管内的发展是腰突症的病理发展过程.有些外在因素,可能导致此过程的加速,使病情加重甚至出现严重并发症.因此如何避免这些不利因素和选择切实可靠的治疗就显得较为重要.我们从1992～2000年间收治了92例最后由手术证实为破裂型腰椎间盘突出症患者.并对这些病例的临床病史特征进行分析,以探讨可能导致纤维环全层破裂的相关因素.     

三维牵引治疗腰椎间盘突出症

九十年代中期以来,三维牵引作为保守治疗腰椎间盘突出症(LDH)的一种新方法出现[1]。因其成本低廉、安全性高、效益可观,在短时间内得到了迅速推广。但是由于对治疗机理缺乏理解,操作中出现滥用和疗效不佳的现象。我院从1995年1月~2000年1月采用三维牵引为主的方法LDH患者1000例,现将治疗中的一点体会总结如下。     

儿童颈胸椎间盘钙化症的临床诊治

目的：探讨儿童颈胸椎间盘钙化症的临床特点、诊断方法与治疗措施。方法：6例中男5例，女1例；年龄7～14岁，平均10．5岁。X线片均发现有椎间盘钙化，有的钙化间盘组织突入椎管。3例为C4．5，2例C6．7，1例T2．3、T4．5。有反复发作的颈背部疼痛，但无脊髓受损表现。6例均采用对症治疗，包括口服消炎止痛药、理疗、卧床休息、配戴颈围等。结果：症状在治疗后2周～3个月缓解。本组均获得随访，随访时间3-25个月，钙化影在3-12个月消失，平均消失时间7．6个月。结论：X线片是儿童椎间盘钙化症主要诊断方法，椎间隙正常而间盘钙化为重要的诊断与鉴别诊断依据。突入椎管内的实性团块，可经保守治疗而消失。