Effect of osteoporosis and intervertebral disc degeneration on endplate cartilage injury in rats

Objective:To investigate the effect of osteoporosis and intervertebral disc degeneration on the endplate cartilage injury in rats.Methods:A total of 48 female Sprague Dawley rats(3 months)were randomly divided into Groups A,B,C and D with 12 rats in each group.Osteoporosis and intervertebral disc degeneration composite model,simple degeneration model and simple osteoporosis model were prepared in Groups A,B and C respectively.After modeling,four rats of each group at 12th.18th and 24th week were sacrificed,Intervertebral height of cervical vertebra C6/C7 was measured.Micro-CT was used to image the endplate of cephalic and caudal cartilage at C6/C7 intervertebral disc.Abraded area rate of C6 caudal and C7 cephalic cartilage endplate was calculated,and then C6/C7 intervertebral disc was routinely embedded and sectioned.stained with safranin O to observe histological changes microscopically.Results:At 12,18 and24 weeks,intervertebral disc height of C6/C7 were(0.58±0.09)mm,(0.53±0.04)mm and(0.04±0.06)mm in Group A rats,(0.55±0.05)mm,(0.52±0.07)mm and(0.07±0.05)mm in Group B rats.At 24th week.intervertebral disc height of Group A rats was significantly lower than that of Group B rats(P0.05);intervertebral disc height of Groups A and B rats at each time point were significantly lower than that of Groups C and D(P0.05).There was no significantly statistical difference of intervertebral disc height between Groups C and D(P0.05).At 12 and 18 weeks,the abraded rate of C6 caudal and C7 cephalic cartilage endplate in Group A rats were significantly higher than that in Groups B.C and D rats(P0.05);the abraded rate in Group B was significantly higher than that in Groups C and D(P0.05).Microscopic observation of CT showed that ventral defects in C6caudal or C7 cephalic cartilage endplate in Groups A and B appeared after 12 weeks of modeling;obvious cracks were found in front of the C6 and C7 vertebral body,and cartilage defect shown the trend of"repairing"at 18 and 24 weeks after modeling.Conclusions:Intervertebral disc degeneration and osteoporosis can cause damage to the cartilage endplate.Co-existence of these two factors can induce more serious damage to the endplate.which has possitive correlation with intervertebral disc degeneration.Osteoporosis plays a certain role in intervertebral disc degeneration process,and accelerates the degeneration of intervertebral disc in a specific time window.     

椎间盘退变与终板内微血管形态改变的相关性研究

目的:通过观测大白兔椎间盘退变过程中椎间盘终板内的血管形态以及血流量的改变,探讨终板内微血管的改变与椎间盘退变之间的相关性。方法:选用40只新西兰大白兔随机分为2组,通过切除造模组20只免腰椎棘间、棘上韧带及棘突、关节突,造成力学失稳状态诱导形成椎间盘退变模型。分别在术后4、8个月通过扫描电镜、血流激光多普勒仪测定椎体终板内的血管形态以及血流量。结果:在椎间盘退变过程中,椎间盘终板内的血管芽形态逐渐被破坏,微血管数量相应减少,终板内的血流量也明显减少,同时终板内血流量中心部位(靠近髓核区域)血流量多于终板内周围区域的血流量。结论:椎体终板内微血管的改变可能是椎间盘退变的促进因素。     

Spondylosis in sand rats: a model of intervertebral disc degeneration and hyperostosis

This study defines gross, histopathologic, and radiologic changes associated with intervertebral disc degeneration in a spontaneously occurring form of the disease in aging sand rats (Psammomys obesus). Sand rats (male/female) fed lab chow supplemented with desert salt bush were sacrificed at periods of 3-30 months. Lateral thoracolumbar spine films were obtained. At sacrifice, spines were surgically exposed and gross findings were recorded; after fixation/decalcification, histopathologic studies were carried out using hematoxylin and eosin, and Safranin-O with fast green counterstain. Metabolic studies included correlations of pathologic and radiologic findings with blood glucose and insulin levels. Disc-space narrowing and subchondral endplate sclerosis increased radiologically with age, with more severe lower lumbar disc lesions. Ligamentous calcifications ventral to involved discs and caudal vertebrae were common. Disc thinning and anterior vertebral bony/cartilaginous spurs were more marked with age. Microscopy revealed loss of nucleus pulposus physaliform cells, chondrocyte replication, disc necrosis, and ossification. Hyperglycemia with and without hyperinsulinemia was common. No statistically significant differences in pathologic findings were noted, neither in diabetic versus nondiabetic nor in hyperinsulinemic animals. The sand rat is a model of disc degeneration; similarities with possible overlap with diffuse idiopathic skeletal hyperostosis syndrome were noted.     

侧前方腰椎椎体间融合术的研究进展

腰椎椎体间植骨融合术是治疗腰椎椎间盘退变性疾病(disc degenerative disease,DDD)的经典手术方式,包括后路腰椎椎体间融合术(posterior lumbar interbody fusion,PLIF)、经椎间孔入路腰椎椎体间融合术(transforaminal lumbar interbody fusion,TLIF)、极外侧入路腰椎椎体间融合术(extreme lateral interbody fusion,XLIF)。     

基于基因表达汇编数据库的外周血中脊柱椎间盘退行性变诊断标志物的生物信息学分析

目的 通过生物信息学分析椎间盘退行性变（IDD）相关的差异表达基因（DEG），寻找疾病的新型诊断标志物。方法 通过基因表达汇编（GEO）数据库GSE124272、GSE150408数据集下载IDD相关的外周血样本芯片数据，筛选出IDD组和正常组之间的DEG。使用DAVID在线数据库对DEG进行基因本体（GO）功能富集和京都基因与基因组百科全书（KEGG）信号通路富集，然后利用STRING在线数据库和Cytoscape软件构建蛋白质-蛋白质相互作用（PPI）网络并获取关键基因，并利用GSE23130数据集中的纤维环样本芯片数据进行验证。利用GSE124272、GSE150408数据集中的数据，采用受试者工作特征（ROC）曲线评估外周血中关键基因的诊断效能。结果 联合分析后筛选出597个DEG，包含363个上调基因和234个下调基因。GO功能富集分析发现DEG主要参与细胞黏附、细胞凋亡、趋化作用和细胞迁移等功能，KEGG分析发现DEG主要参与细胞外基质受体相互作用和癌症中的信号通路。PPI网络分析筛选出17个关键基因，经验证获得RBMX、EEF1A1、SSR1和POLR2C这4个基因，ROC曲线分析显示这4个基因对IDD诊断效能显著，曲线下面积分别为0.763、0.741、0.710、0.702。结论 RBMX、EEF1A1、SSR1和POLR2C或可成为IDD的新型诊断标志物，为该病进一步的功能研究提供理论依据。     

Clinical and radiographic features of spinal osteoarthritis predict long-term persistence and severity of back pain in older adults

BackgroundPatients with back pain can show one or more features of spinal osteoarthritis (OA), such as morning stiffness, limited or painful range of motion (ROM), and lumbar disc degeneration (LDD). However, it has not been investigated whether these features are prognostic of long-term back pain.ObjectivesThis study assessed whether spinal morning stiffness, ROM and LDD are prognostic factors for back pain after 1 year in older adults with back pain.MethodsThis prospective observational study (BACE cohort) included patients aged > 55 years visiting a general practitioner for a back-pain episode. Baseline patient-reported morning stiffness, physical examined ROM and radiographic LDD features (i.e., multilevel osteophytes and disc space narrowing) were analysed as potential prognostic factors in unadjusted and adjusted regression models with the outcomes of persistent back pain (yes/no) and back pain severity after 1-year follow-up.ResultsThis study included 543 patients with mean (SD) age 67 (8) years, 59% female, and 62% reporting back pain at 1-year follow-up. When studied in separate adjusted models, persistent back pain was associated with morning stiffness > 30 min (OR 3.0, 95%CI 1.3; 5.5), restricted lateroflexion (OR 1.8, 95%CI 1.0; 3.2), pain during rotation (OR = 1.7, 95%CI 1.0; 2.9), multilevel osteophytes (OR 2.4, 95%CI 1.4; 4.1), and multilevel disc space narrowing (OR 1.5, 95%CI 0.9; 2.4). When investigated in the same adjusted model, persistent back pain remained associated with only morning stiffness > 30 min (OR 2.4, 95%CI 1.0; 3.9), pain during rotation (OR 1.6, 95%CI 0.9; 2.8), and multilevel osteophytes (OR 2.1, 95%CI 1.2; 3.7). The same spinal OA-related features were associated with back pain severity.ConclusionsSpinal morning stiffness, painful rotation, and multilevel osteophytes are prognostic factors for persistent back pain and back pain severity after 1 year. Evaluating these clinical and radiographic features of spinal OA could help clinicians identify older patients who will experience long-term back pain.     

自噬在张力诱导终板软骨细胞退变过程中的变化

 目的 探讨自噬在张力诱导终板软骨细胞退变过程中的变化及作用。方法 取10只清洁级SD大鼠腰椎终板软骨进行细胞培养。对P1代终板软骨细胞分别加载间歇循环张力（10%伸长率，0.5 Hz）0 h、3 h、12 h、24 h、48 h。以倒置相差显微镜观察细胞形态学变化，实时PCR与蛋白印迹法检测软骨标志基因Ⅱ型胶原、转录因子SOX-9及蛋白多糖转录因子、Beclin-1和LC3基因表达的变化，以单丹磺酰戊二胺染色观察自噬小体。MTT（3-2，5-二苯基四氮唑溴盐染色）法检测3-甲基腺嘌呤（自噬抑制剂）刺激前后的细胞存活率。结果 间歇循环张力诱导后0 h组与3 h组为正常终板软骨细胞形态，呈多角形；12 h组略呈不规则形；24 h组和48 h组呈梭形改变。实时PCR显示24 h组和48 h组中Ⅱ型胶原、转录因子SOX-9及蛋白多糖的表达量降低；自噬相关基因LC3和Beclin-1表达量呈时间依赖性增加。单丹磺酰戊二胺染色显示24 h组和48 h组自噬发生率呈时间依赖性增加。MTT结果显示细胞存活率呈降低趋势；添加3-甲基腺嘌呤刺激后细胞活性减弱、存活率降低。结论 间歇循环张力刺激下终板软骨细胞表型逐渐丧失；自噬相关基因LC3与Beclin-1表达明显上调，但细胞活性降低；抑制自噬水平可降低细胞存活率，提示自噬参与了间歇循环张力诱导的终板软骨细胞退变过程。     

纤维环针刺法建立兔椎间盘退变模型的实验研究

目的建立一种稳定可靠、可重复性好、成功率高，且与人类退变相似的椎间盘退变模型。方法38只普通级新西兰大白兔，雌雄不计，随机选取2只动物用于研究兔腰椎的解剖学特点，另外36只动物用随机数字表法均分为模型组和对照组。模型组手术取右侧腹膜后入路，暴露至L5/L6、L4/L5椎间盘纤维环，予16G穿刺针针刺纤维环；对照组同法暴露不进行针刺。分别于术前及术后4周对实验动物行X线和MRI检查，术后4周处死实验动物，取椎间盘标本行病理学检查。结果术后4周，X线和MRI检查显示模型组椎间盘高度明显降低、部分骨赘形成、终板硬化，T2加权像信号较术前明显降低、椎间盘颜色变黑变暗，髓核缩小或消失，纤维环变厚；而对照组椎间盘未见明显变化，T2加权像呈均匀高信号；取椎间盘标本经HE染色，电镜下见模型组髓核组织明显缩小，结构紊乱，细胞密度较小，纤维向内塌陷，纤维环板层出现明显断裂，排列不整齐，纤维环内层见少量毛细血管长入；对照组腰椎间盘病理学检查无明显退变表现；免疫组化见模型组椎间盘内TNF-a数量较对照组明显增多。结论该方法具有操作简单、可行、高效等优点，是建立椎间盘退变模型的一种可靠方法。