腰椎间盘中血管内皮生长因子的表达及其意义

目的 :观察并探讨腰椎间盘中内源性血管内皮生长因子 (vascularendothelialgrowthfactor,VEGF)的表达分布规律及其意义。方法 :采用免疫组织化学技术检测 1 6例胎儿期、8例生长期、1 2例成熟退变期和 42例突出椎间盘中VEGF的表达。结果 :胎儿椎间盘脊索细胞和纤维环外层血管内皮细胞出现阳性免疫染色 ,阳性率为 87 5 % ;生长期和成人期椎间盘未见阳性表达 ;退变突出组总阳性率为 83 3 %。VEGF表达主要出现于破裂型和游离型椎间盘突出 (P <0 0 1 ) ,其细胞来源主要是突出椎间盘组织中毛细血管内皮细胞、髓核内的类软骨细胞及单核巨噬细胞。年龄与VEGF表达阳性率关系不大 (P >0 0 5)。病程超过 1年者VEGF阳性率明显低于 1年以内者 (P <0 0 5)。结论 :胎儿期和破裂型 (包括游离型 )突出椎间盘组织可产生内源性VEGF ,突出椎间盘中VEGF的阳性表达与病程、突出类型密切相关     

Expression of Fas receptor on disc cells in herniated lumbar disc tissue

STUDY DESIGN: The expression of Fas receptor, an apoptosis-related protein, on disc cells was examined in surgically obtained disc specimens. OBJECTIVE: To assess the fate of disc cells in herniated disc tissue and the difference in the degree of expression of the Fas receptor between contained and noncontained discs. SUMMARY OF BACKGROUND DATA: Little is known about the fate of disc cells after herniation. METHODS: Twenty-three herniated lumbar disc specimens were classified into contained discs (protrusion or subligamentous extrusion; n = 9) and noncontained discs (transligamentous extrusion or sequestration; n = 14). All specimens were stained using the avidin-biotin-peroxidase complex method. The percentage of disc cells positive for Fas receptor was calculated and compared with clinical and radiologic data. RESULTS: There was a significant difference in the percentage of Fas-positive disc cells between the contained and noncontained discs (8.44 vs.- 14.29;P = 0.044). The percentage of Fas-positive disc cells correlated significantly with the patient's age (r = 0.455, P = 0.029), but not with the degree of disc degeneration on magnetic resonance imaging (r = 0.252, P = 0.214). CONCLUSION: This is the first study to identify the expression of Fas receptor on disc cells in herniated disc tissue. The results show that the disc cells after herniation may undergo Fas-mediated apoptosis and that the degree of expression of Fas receptor differs depending on the type of herniation.     

基质金属蛋白酶-3和其组织抑制剂-1在椎间盘中的表达及其意义

目的：检测突出和非突出椎间盘中是否有基质金属蛋白酶－3（MMP3）及其组织抑制剂－1（TIMP）的表达,了解椎间盘退变和突出的发病机制。方法：采用ABC免疫组化方法,测定60例突出椎间盘标本（分为凸出型,脱出型,游离型）和16例非突出椎间盘标本内MMP3和TIMP1的表达情况,结果：突出椎间盘中的MMP3比非突出椎间盘的多,其差异有显著性意义。脱出型及游型内的MMP3比突出型的多,其差异有显著性意义。脱出突型及游离型内的MMP3无显著性差异,非突出椎间盘和凸出型椎间盘内TIMP1为阴性,脱出型和游离内TIMP1为阳性,但无显著性差异,结论：MMP3和TIMP1的不平衡表达也许是椎间盘退变的因素,椎间盘退变程度的差异可能是椎间盘突出症临床分型的基础。     

肥大细胞和巨噬细胞在疼痛椎间盘的分布和意义

目的研究肥大细胞和巨噬细胞在疼痛椎间盘的分布并探讨其与椎间盘退变的关系。方法收集腰椎后路切除的15个椎间盘源性下腰痛病人的21个通过腰椎间盘造影术证实的疼痛椎间盘,同时收集16个在M RIT2加权上信号强度明显减弱的无腰痛症状的生理老化椎间盘和10个正常对照椎间盘,行组织学检查并用免疫组化方法观察肥大细胞和巨噬细胞在不同椎间盘的分布。结果免疫组织化学结果显示在疼痛椎间盘的肉芽组织区有大量的肥大细胞和巨噬细胞分布,在肉芽组织邻近区有少量分布,在非肉芽组织区、生理老化椎间盘和正常对照椎间盘没有分布。结论研究结果提示肥大细胞和巨噬细胞在纤维环外层损伤后激发的炎症反应和随之的椎间盘退变过程中起关键作用。     

重度肩锁关节脱位的手术治疗

目的：对15例重度肩锁关节脱位的手术7治疗进行治疗,方法：15例全部为Allman分型中的Ⅲ型损伤,其中有11例切除纤维软骨盘,7例修复锁韧带,3例加用一枚松质骨螺钉固定于锁骨与喙空间,肩锁韧带全部修复,用两枚克氏针交叉固定于肩锁关节。结果：经10个月－6年的随访。疗效评价按Karlsson分类A级为12例,B级3例,病人均重返原工作岗位,有10例喙锁间隙软组织钙化,但对病人肩关节活动并无影响,结论：对重度肩锁关工锐位的病人应尽早手术治疗。克氏针交叉内固定是一种简单而有效的方法,纤维软骨盘是否切除和喙锁韧带是否修复对预后无明显影响。     

Histology of intervertebral disc protrusion: an experimental study using an aged rat model

STUDY DESIGN: In vitro experimental intervertebral disc ruptures of aged rats were examined histologically. OBJECTIVES: To clarify the mechanism of intervertebral disc herniations by microscopic investigation of ruptured discs. SUMMARY OF BACKGROUND DATA: Clinically, disc herniations have been classified into two types: extrusion and protrusion. However, the pathogenesis of protrusion type herniations has not yet been demonstrated by any studies. To clarify this issue, it is essential to establish an appropriate model producing disc herniations, and to examine the sequential changes in the structure of herniated discs. METHODS: Lumbar discs of 2-year-old rats were examined histologically and compared with human lumbar discs. To examine structural changes in discs subjected to repetitive motion stress, 400 repetitions of a sequence of flexion (30 degrees ) and axial rotation (6 degrees ) were applied in vitro to the lumbar discs of the animals. RESULTS: The microstructure of normal lumbar discs in aged rats was similar in many ways to the human lumbar discs in a 20- to 40-year-old adult. Of 10 discs subjected to repetitive stress, 4 were ruptured at the junction between the posterior anulus fibrosus and the sacral cartilage endplate. One had an extruded nucleus pulposus, and three had a protruded anulus fibrosus, which displayed disorganized structure containing widened and flaccid lamellae. CONCLUSIONS: The results from this study indicate that disc protrusion can be caused by disorganization of the ruptured annular lamellae, not by focal compression of the nucleus pulposus.     

Expression of Fas ligand and apoptosis of disc cells in herniated lumbar disc tissue

STUDY DESIGN: An examination of surgically obtained herniated lumbar disc tissues performed by using immunohistochemical staining and the DNA nick end labeling method. OBJECTIVE: To investigate the cell type that expresses Fas ligand (FasL) and any evidence of apoptosis of the disc cells in herniated lumbar disc tissues. SUMMARY OF BACKGROUND DATA: The Fas/FasL system is involved in delivering a death signal that rapidly commits the cells to apoptosis. In the authors' previous study, the expression of Fas on disc cells was identified in herniated lumbar disc tissue. METHODS: Twenty-three herniated lumbar disc tissues (contained disc, n = 9; noncontained disc, n = 14) were examined to investigate the cell type that expresses FasL and any evidence of apoptosis of the disc cells by using immunohistochemical staining and the DNA nick end labeling method, respectively. The percentage of FasL-positive disc cells was calculated and compared with clinical and radiologic data. RESULTS: FasL was expressed in the cytoplasm of the disc cells, and nuclear DNA fragmentation in a few disc cells was identified. A higher degree of FasL expression in disc cells was found in noncontained discs than in contained discs (P < 0.05). The percentage of FasL-positive disc cells significantly increased with the patient's age (P < 0.05), but not with the degree of disc degeneration (P > 0.05). CONCLUSION: The current results indicate that disc cells, after herniation, undergo apoptotic cell death via autocrine or paracrine FasL mechanisms by the disc cells themselves.     

The role of mast cells in disc herniation inflammation.

STUDY DESIGN: A study of herniated lumbar disc tissue samples and control disc material to determine the presence of mast cells in disc herniations. OBJECTIVES: To analyze whether mast cells have any involvement in disc herniation pathophysiology and lumbar pain, because mast cells may have an important role in acute and chronic inflammatory responses. SUMMARY OF BACKGROUND DATA: Studies of inflammatory cells, biochemical mediators of inflammation, and tissue degrading enzymes have suggested that these factors may be involved--and perhaps play an important role--in the pathophysiology of lumbar pain and radiculopathy. Mast cells are known to play an important role in acute and chronic inflammatory responses. It was therefore of interest to clarify their possible role in intervertebral disc herniation inflammation. METHODS: Fifty herniated lumbar discs from 50 patients who had undergone disc surgery and three normal control discs were obtained. Sections from every disc then were examined histologically and immunocytochemically for mast cells by using monoclonal antibodies to either of two types of specific proteases of mast cells, tryptase and chymase. RESULTS: By none of the methods could any mast cells be observed in any of the control disc samples. With toluidine blue staining, mast cells were observed in 9 of 50 (18%) of discs. Mast cells immunoreactive to either tryptase or chymase were observed in 10 of 50 disc samples (20%) and immunoreactive for tryptase and chymase simultaneously in 4 of 50 disc samples (8%). However, the majority of the samples studied (80%) demonstrated immunoreactivity to neither tryptase nor chymase. Among the samples studied were five disc protrusions that totally lacked mast cells. CONCLUSIONS: A minority of disc herniations exhibited mast cells, as verified by toluidine blue staining and immunocytochemistry. The results may suggest a role of mast cells in intervertebral disc herniation inflammation, but only in a subset of these cases. Massive infiltration by mast cells never was observed.     

人退变腰椎间盘中细胞凋亡与Bax及半胱胺酸蛋白酶蛋白3基因的表达

目的 检测人腰椎间盘组织中的细胞密度、细胞凋亡率以及Bax和半胱氨酸蛋白酶蛋白3(Caspase-3)的表达,为深入了解人腰椎间盘退变的机制提供实验依据,并为将来通过生物学方法 延缓腰椎间盘退变提供新的思路.方法 实验组30个椎间盘标本(L2～S1)来自27例行后路腰椎间盘切除椎间融合手术自愿捐赠的患者,男18例,女9例;年龄30～72岁,平均51.09岁.所有病变节段均经MRI证实,患者术前均未接受椎间盘造影、胶原酶髓核溶解或椎间盘激光汽化术.对照组20个标本(L2～S1)来自5例青年男性意外死亡者新鲜尸检腰椎间盘;年龄24～37岁,平均30.83岁.采用HE染色观察凋亡软骨细胞、凋亡小体和细胞密度,TUNEL染色法检测人腰椎间盘中的凋亡细胞率,用SP免疫组织化学法检测Bax及Caspase-3表达,图像分析Bax及Caspase-3的平均灰度值.结果 HE染色示对照组、实验组软骨终板和髓核内的平均细胞密度分别为(17.16±1.22)、(12.41±0.95)个/HP,二者差异有统计学意义(P＜0.01).TUNEL染色观察示对照组的软骨终板与髓核内的平均凋亡细胞率为6.97%±0.92%,低于实验组的12.59±0.95%(P＜0.01).SP免疫组织化学染色示对照组髓核内Bax、Caspase-3染色阳性细胞率分别为11.02%4±1.18%和9.01%±1.00%,均低于实验组的19.29%±1.18%和15.07%±0.97%,差异均有统计学意义(P＜0.01).对照组腰椎间盘髓核内Bax、Caspase-3的平均灰度值分别为187.33±7.88和185.68±3.26,实验组分别为124.98±6.69和160.13±4.37,两组间比较差异有统计学意义(P＜0.01).将全部腰椎间盘标本进行Pearson相关分析,对照组和实验组腰椎间盘凋亡细胞率与细胞密度间成负相关,相关系数r分别为-0.88和-0.93(P＜0.01)两组髓核内Bax、Caspase-3阳性细胞率与凋亡细胞率之间成正相关,相关系数r分别为0.83和0.91(P＜0.01).结论 细胞密度下降参与了人腰椎间盘的退变过程,Bax和Caspase-3的表达上调在人腰椎间盘髓核细胞的凋亡过程中有一定作用.     

Polarization of infiltrating macrophages in the outer annulus fibrosus layer associated with the process of intervertebral disc degeneration and neural ingrowth in the human cervical spine

BACKGROUND CONTEXTAs no infiltrating macrophages exist in healthy discs, understanding the role of infiltrating macrophages including their polarity (M1 and M2 phenotypes) in intervertebral discs (IVDs) is important in the assessment of the pathomechanisms of disc degeneration.PURPOSETo determine the relationship between infiltrating macrophage polarization and the progression of human cervical IVD degeneration.STUDY DESIGNHistopathological study using harvested human cervical IVDs.METHODSIVDs collected during anterior cervical decompression from 60 patients were subjected to immunostaining and immunoblotting. The samples were classified as type 0–3 according to the percentage of CD16- and CD206-positive cells to CD68-positive cells in the outer annulus fibrosus layer. The number of vessels and nerve fibers and the severity of chronic inflammation with a focus on inflammatory cell infiltration, fibrosis, and capillary proliferation were also assessed.RESULTSThe number of CD16-positive cells was the highest in type 2 IVDs, and was suppressed following the infiltration of CD206-positive cells. The degree of chronic inflammation was significantly higher in type 2 and type 3 IVDs, and the number of nerve fibers was significantly higher in type 3 IVDs. The endothelial cells of small vessels were positive for nerve growth factor, brain-derived neurotrophic factor, and neurotrophin-3 expression. Staining for tropomyosin receptor kinase (Trk)-A, Trk-B, and Trk-C was positive in aberrant fibers. In immunoblot analysis, the expression levels of these neurotrophic factors and receptors were significantly higher in type 2 and 3 IVDs.CONCLUSIONSThe polarity of macrophages around newly developed microvasculature might be altered with cervical IVD degeneration. A higher number of infiltrating M1 macrophages around the vessels was associated with chronic inflammation; however, their number got suppressed following the infiltration of M2 macrophages. The expression of neurotrophins in the capillaries of small vessels might contribute to neural ingrowth into degenerated IVDs.CLINICAL SIGNIFICANCEClarifying macrophages polarity change around new microvasculature associated with progression of IVD degeneration could enhance our understanding of the underlying mechanisms of neural ingrowth into degenerated IVDs and lead to development of a novel therapeutic target for prevention of IVD.     

退变颈椎间盘中IL-17表达与分布的研究

目的 观察退变颈椎间盘中自细胞介素-17(IL-17)的表达与分布,并探讨其与颈椎间盘退变发生发展的关系.方法 实时荧光相对定量PCR(RQ-PCR)检测30例退变颈椎间盘及10例正常对照椎间盘中IL-1β、IL-17、肿瘤坏死因子-α(TNF-α)和孤核受体(retinoid-related orphan re-ce...     

Gene expression of collagen types IX and X in the lumbar disc

To study gene expression of collagen typesIX and X in human lumbar intervertebral discs duringaing and degeneration and to explore the role of collagentypes IX and X in disc degeneration.     

Fas ligand expression on human nucleus pulposus cells decreases with disc degeneration processes

Background The intervertebral disc has been reported to be an immunologically privileged environment, possibly mediated by Fas ligand (FasL) expression. On the other hand, recent studies have shown the infiltration of host immune cells into the degenerated disc, which may indicate the failure of the immune-privilege feature of the disc with degeneration. However, the relationship between FasL expression and disc degeneration is still unclear. Therefore, the purpose of this study was to clarify the relationship between FasL expression and disc degeneration. Methods Ten human degenerated disc specimens were obtained from spondylolisthesis patients and ten nondegenerated discs from idiopathic scoliosis patients during surgical procedures. Immunohistochemical staining was performed to determine the presence of FasL in cross-sections of those discs. Parts of the disc tissues were used to examine FasL expression quantitatively with Western blot analysis. To examine whether the change in FasL expression was influenced by aging, an animal study comparing the discs from young and old rats were performed using magnetic resonance imaging (MRI) and real-time polymerase chain reaction (PCR) assessment. Results Nucleus pulposus cells showed strong positive staining for FasL in all specimens examined. Quantitative examination demonstrated a significant decrease in FasL expression in the degenerated group compared with the nondegenerated group (average 67.6%, P<0.05). MRI showed no significant differences in the grade of disc degeneration between young and old rats, and also no significant difference in FasL mRNA in real-time PCR assay. Conclusions The current results indicate that FasL and its potential mechanism of immunological privilege could influence the protection of the intervertebral disc against degeneration.     

Immunohistochemical identification of notochordal markers in cells in the aging human lumbar intervertebral disc

The fate of notochord cells during disc development and aging is still a subject of debate. Cells with the typical notochordal morphology disappear from the disc within the first decade of life. However, the pure morphologic differentiation of notochordal from non-notochordal disc cells can be difficult, prompting the use of cellular markers. Previous reports on these notochordal cell markers only explored the occurrence in young age groups without considering changes during disc degeneration. The aim of this study, therefore, was to investigate presence, localization, and abundance of cells expressing notochordal cell markers in human lumbar discs during disc development and degeneration. Based on pilot studies, cytokeratins CK-8, -18 and -19 as well as Galectin-3 were chosen from a broad panel of potential notochordal cell markers and used for immunohistochemical staining of 30 human lumbar autopsy samples (0–86 years) and 38 human surgical disc samples (26–69 years). In the autopsy group, 80% of fetal to adolescent discs (0–17 years) and 100% of young adult discs (18–30 years) contained many cells with positive labeling. These cells were strongly clustered and nearly exclusively located in areas with granular changes (or other matrix defects), showing predominantly a chondrocytic morphology as well as (in a much lesser extent) a fibrocytic phenotype. In mature discs (31–60 years) and elderly discs (≥60 years) only 25 and 22–33%, respectively, contained few stained nuclear cells, mostly associated with matrix defects. In the surgical group, only 16% of samples from young adults (≤47 years) exhibited positively labeled cells whereas mature to old surgical discs (>47 years) contained no labeled cells. This is the first study describing the presence and temporo-spatial localization of cells expressing notochordal cell markers in human lumbar intervertebral discs of all ages and variable degree of disc degeneration. Our findings indicate that cells with a (immunohistochemically) notochord-like phenotype are present in a considerable fraction of adult lumbar intervertebral discs. The presence of these cells is associated with distinct features of (early) age-related disc degeneration, particularly with granular matrix changes.     

Reevaluation of discograms not classified into usual classifications

Discograms of images that were eccentrically dyed because of insufficient infiltration of contrast medium are difficult to classify into the usual past discogram patterns. In this study, these types of images were detected in 40 discs of 36 patients with lumbar disc disease. We classified these images into the following three types, and analyzed the dye mechanisms in each case by computed tomography discographic findings: (1) type A (image of the annulus fibrosus only). Nine discs in nine cases. A part of the marginal annulus fibrosus was dyed. (2) type B (image of the right or left half of the nucleus pulposus). Eighteen discs in 15 cases. Unilateral dyeing was considered nucleus pulposus existing in the central region of the disc. (3) type C (partial image of the superior or inferior half of the nucleus pulposus). Thirteen discs in 12 cases. Only the superior or inferior half showing a cotton-ball pattern was dyed.     

Immunophenotypic analysis of the inflammatory infiltrates in herniated intervertebral discs

STUDY DESIGN: The herniated portion of the lumbar disc was analyzed immunohistochemically for inflammatory infiltrates to determine their immunophenotype. OBJECTIVE: To investigate the pathomechanism behind spontaneous regression of herniated discs. SUMMARY OF BACKGROUND DATA: Spontaneous regression of herniated intervertebral discs has been increasingly reported. The inflammatory response of the host has been suggested as a factor in this phenomenon. However, whether the inflammation is induced from direct chemical irritation of the nucleus pulposus material or whether it is secondary to an autoimmune response to the nucleus pulposus remains controversial. METHODS: The herniated portion of the disc was collected from 38 patients who underwent surgery for lumbar disc herniation. Thin cryostat sections were made, and the extent to which inflammatory cells had infiltrated the disc specimen was defined. Then the immunophenotype of cellular infiltrates in the herniated disc specimens was assessed by immunostaining using a series of antibodies for lymphocyte, monocyte, macrophage, and dendritic cell markers. RESULTS: The inflammatory infiltrates in 14 of the 38 herniated discs were subjected to immunohistochemical analysis. None of them expressed the immunophenotypic markers of the lymphocyte (CD20, CD45RO, CD4, CD8, TCRgammadelta), mature monocyte (CD33), or dendritic cell (CD1a, CD80, CD86, S100). Abundant infiltration of CD68-positive cells that lacked CD33 but had a variable amount of CD11b, CD11c, and CD40 likely represents a process of differentiation from monocytes to macrophages. CONCLUSIONS: These findings are consistent with an immunophenotype of inflammatory responses to tissue injury or chemical irritation rather than antigen-specific immune responses. Therefore, understanding the mechanism of tissue repair is fundamentally important in the management of patients with disc herniations.     

椎间盘源性下腰痛的发病机制

目的探讨椎间盘源性下腰痛的发病机制。方法收集腰椎后路切除的17例椎间盘源性下腰痛患者的19个经腰椎间盘造影术证实的疼痛腰椎间盘;同时收集12个在MRI T2加权像上信号强度明显减弱、无腰痛症状的生理老化椎间盘和10个正常对照椎间盘,行组织学检查和P物质、神经丝蛋白和血管活性肠肽的免疫组织化学染色检查。结果椎间盘源性下腰痛患者的疼痛椎间盘在组织学上的显著特征表现为,一条从髓核至纤维环外层的血管化肉芽组织条带区,其间伴有1个或多个裂隙;肉芽组织条带区与椎间盘造影术后CT上显示的纤维环裂隙一致,肉芽组织之外的纤维环结构基本正常。生理老化椎间盘和正常对照椎间盘表现为与年龄相关的改变。免疫组织化学染色显示,疼痛椎间盘中P物质、神经丝蛋白和血管活性肠肽3种神经肽阳性神经纤维分布数量和比例,较正常对照椎间盘和生理老化椎间盘明显增多;神经纤维主要沿伴有裂隙的肉芽组织条带区分布;疼痛椎间盘髓核中可见P物质和神经丝蛋白的阳性神经纤维分布。结论椎间盘后方神经分布广泛的肉芽组织条带区是椎间盘造影术疼痛和椎间盘源性下腰痛的起源部位。肉芽组织条带可能起源于椎间盘的创伤修复过程。生理老化椎间盘和疼痛椎间盘的差异是后者形成组织学上的肉芽组织条带区。     

Human intervertebral disc cells are genetically modifiable by adenovirus-mediated gene transfer: implications for the clinical management of intervertebral disc disorders

STUDY DESIGN: Human intervertebral disc cells were cultured in monolayer and treated with adenovirus-containing marker genes to determine the susceptibility of the cells to adenovirus-mediated gene transfer. OBJECTIVES: To test the efficacy of the adenovirus-mediated gene transfer technique for transferring exogenous genes to human intervertebral disc cells in vitro. SUMMARY OF BACKGROUND DATA: Upregulated proteoglycan synthesis after direct in vivo adenovirus-mediated transfer of growth factor genes to the rabbit intervertebral disc has previously been reported. Before contemplating extending this approach to the treatment of human disc disease, it is necessary to demonstrate that human intervertebral disc cells are indeed susceptible to adenovirus-mediated gene transduction. METHODS: Human intervertebral disc cells were isolated from disc tissue obtained from 15 patients during surgical disc procedures. The cells were cultured in monolayer and treated with saline containing five different doses of adenovirus carrying the lacZ gene (Ad/CMV-lacZ), saline containing adenovirus carrying the luciferase gene (Ad/CMV-luciferase), or saline alone. Transgene expression was analyzed by 5-bromo-4-chloro-3-indolyl-beta-galactosidase (X-Gal) staining and luciferase assay. RESULTS: Adenovirus efficiently transferred lacZ and luciferase marker genes to cells from degenerated discs as well as to cells from nondegenerated discs. A minimum dose of 150 MOI Ad/CMV-lacZ was found to be sufficient to achieve transduction of approximately 100% of disc cells-regardless of patient age, sex, surgical indication, disc level, and degeneration grade. No statistically significant difference in the luciferase activities could be detected in disc cell cultures from degenerated and nondegenerated discs treated with Ad/CMV-luciferase. CONCLUSIONS: In vitro transducibility of human intervertebral disc cells by adenovirus is relatively insensitive to disc degeneration grade. Because the rate-limiting step for successful gene therapy is the ability to transfer genes efficiently to the target tissue, the achievement of efficient gene transfer to human intervertebral disc cells(using a direct, adenovirus-mediated approach) is an important and necessary step in the development of gene therapy strategies for the management of human intervertebral disc disorders.     

细胞凋亡、细胞增殖在人颈椎间盘退变过程中的作用

目的 探讨细胞凋亡、细胞增殖在人颈椎间盘退变过程中的作用以及人颈椎间盘细胞凋亡可能涉及的信号转导途径.方法 收集手术切除的33份突出颈椎间盘组织,以22份正常人颈椎间盘组织作为对照,组织形态学和TUNEL法检测凋亡细胞,免疫组织化学法检测增殖细胞核抗原(PCNA)、bax及Caspase-3的表达.结果 实验组的细胞密度低于对照组(髓核内:6.30±1.54比8.96±1.14;软骨终板内:17.27±1.82比25.41±1.89);实验组的TUNEL阳性细胞率高于对照组(髓核内:11.73±1.36比7.02±1.26;软骨终板内:13.04±1.75PC6.86±1.42);实验组髓核内PCNA阳性细胞率高于对照组(8.38±1.98比4.55±1.54);实验组髓核内bax阳性细胞率和Caspase-3阳性细胞率分别高于对照组(bax:19.32±1.95比10.94±1.72;Caspase-3:15.05±1.74比8.92±1.48);TUNEL阳性细胞率与细胞密度之间呈负相关(P＜0.01);实验组髓核内PCNA阳性细胞率与细胞密度之间呈正相关(P＜0.01);两组髓核内bax阳性细胞率、Caspase-3阳性细胞率均与TUNEL阳性细胞率之间呈正相关(P＜0.01).结论 细胞凋亡与细胞增殖间的不平衡可能是人退变颈椎间盘内细胞密度下降的原因,人颈椎间盘髓核细胞的过度凋亡可能与bax和Caspase-3的表达上调有关.     

Herniated and spondylotic intervertebral discs of the human cervical spine: histological and immunohistological findings in 500 en bloc surgical samples. Laboratory investigation

OBJECT: In this paper the authors' goal was to identify histological and immunohistochemical differences between cervical disc hemrniation and spondylosis. METHODS: A total of 500 cervical intervertebral discs were excised from 364 patients: 198 patients with disc herniation and 166 patients with spondylosis. We examined en bloc samples of endplate-ligament-disc complexes. Types of herniation and graded degrees of disc degeneration on MR images were examined histologically and immunohistochemically. RESULTS: The herniated discs showed granulation tissue, newly developed blood vessels, and massive infiltration of CD68-positive macrophages, which surrounded the herniated tissue mainly in the ruptured outer layer of the anulus fibrosus. The vascular invasion was most significant in uncontained (extruded)-type herniated discs. Chondrocytes positive for matrix metalloproteinase (MMP)-3, tumor necrosis factor (TNF)-alpha, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were abundant in both herniated and spondylotic discs. Free nerve fibers, positive for nerve growth factor (NGF), neurofilament 68, growth-associated protein (GAP)-43, and substance P, were strongly apparent in and around the outer layer of uncontained (extruded)-type herniated discs, with enhanced expression of NGF. The authors observed that herniated discs showed more advanced degeneration in the outer layer of the anulus fibrosus around the granulation tissue than spondylotic discs. On the other hand, spondylotic discs showed more advanced degeneration in the cartilaginous endplate and inner layer of the anulus fibrosus than herniated discs. Spondylotic discs also had thicker bony endplates and expressed TNFalpha and MMP-3 more diffusely than herniated discs, especially in the inner layer of the anulus fibrosus. CONCLUSIONS: The authors' results indicate that herniated and spondylotic intervertebral discs undergo different degenerative processes. It is likely that TNFa, MMP-3, bFGF, and VEGF expression is upregulated via the herniated mass in the herniated intervertebral discs, but by nutritional impairment in the spondylotic discs. Macrophage accumulation around newly formed blood vessels in the herniated disc tissues seemed to be regulated by MMP-3 and TNFalpha expression, and both herniated and spondylotic discs exhibited marked neoangiogenesis associated with increased bFGF and VEGF expression. Nerve fibers were associated with NGF overexpression in the outer layer of the anulus fibrosus as well as in endothelial cells of the small blood vessels.