不同脱细胞方法对猪尾纤维环生物力学特性及组织结构的影响

目的探讨不同脱细胞处理方法对猪尾椎间盘纤维环生物力学特性及组织结构的影响,为构建组织工程纤维环提供实验依据。方法取新鲜猪尾纤维环60个,随机分为4组,每组15个。Triton X-100组(A组):先将纤维环放入Tris-HCl低渗缓冲液中振荡48 h,再用TritonX-100、DNaseⅠ和RNase A对纤维环脱细胞处理;SDS组(B组):将纤维环冻融3次,接着用SDS、DNaseⅠ和RNase A对纤维环脱细胞处理;胰蛋白酶组(C组):用含胰蛋白酶、DNaseⅠ和RNase A的Tris缓冲液对纤维环脱细胞处理;对照组(D组):纤维环不做任何处理。采用HE染色与扫描电镜方法,观察各组脱细胞情况以及纤维环超微结构的变化;采用生物化学和生物力学方法检测各组纤维环的胶原含量、氨基葡聚糖(GAG)含量及力学参数。结果与对照组比较,HE染色及扫描电镜可见A、B、C 3组均无细胞残留;A组纤维环超微结构未见破坏,C组纤维环超微结构可见轻度破坏,B组纤维环超微结构可见明显破坏。A、B、C 3组纤维环胶原含量与对照组比较无统计学差异(P>0.05),但GAG含量均明显低于对照组(P<0.05)。A、C两组的力学参数(极限载荷、极限应力、韧度、弹性模量、断裂功耗)与对照组无统计学差异(P>0.05),B组上述力学参数均低于对照组(P<0.05)。结论 Triton X-100组处理后的猪尾椎间盘纤维环无细胞残留,结构无破坏,基质成分及力学性能保存良好,适合于构建组织工程纤维环。     

Effects of swelling pressure and hydraulic permeability on dynamic compressive behavior of lumbar annulus fibrosus

The objective of this study was to investigate the effects of swelling pressure and hydraulic permeability on the dynamic behavior of intervertebral disk tissue in confined compression. Normal (served as a control) and trypsin-treated, axial annulus fibrosus (AF) specimens from the porcine lumbar disks were tested and their swelling strain, swelling pressure, equilibrium compressive modulus (H _A, dynamic modulus, and hydraulic permeability (k) were determined at 30% and 40% swelling strain levels. The proteoglycan depletion due to trypsin treatment resulted in significantly lower values of the free swelling strain, swelling pressure, equilibrium modulus, dynamic modulus, and higher value of hydraulic permeability for trypsin-treated group, comparing to those for the control group. At the 30% swelling strain level, the equilibrium moduli were 51.84±14.53 kPa (n=8) for the control group and 15.11±5.67 kPa (n=8) for the trypsin-treated group; and the hydraulic permeabilities were 4.50E-15±1.60E-15 m⁴/Ns and 8.43E-15±4.29E-15 m⁴/Ns for control and trypsin-treated groups, respectively. No statistically significant difference in wet tissue density or dry tissue density was found between control and trypsin-treated groups. There was a significant correlation between swelling pressure and compressive (aggregate) modulus (R²=0.824, m=22). The decrease in measured dynamic modulus for trypsin-treated group was attributed to the reduced swelling pressure (or modulus H _A and increased hydraulic permeability (k) due to PG depletion. © 2002 Biomedical Engineering Society. PAC2002: 8719Rr, 8719Tt, 8360Jk     

黄芪多糖对颈椎病模型大鼠颈椎间盘纤维环MMP2和MMP9表达的影响

目的:探究黄芪多糖(AP)对颈椎病模型大鼠颈椎间盘纤维环中基质金属蛋白酶2(MMP2)和MMP9表达的影响。方法:建立动静力失衡性颈椎间盘退变大鼠模型,将造模成功大鼠随机分为模型组(M组)及AP低、高剂量处理组(L-AP组和H-AP组),以假手术组大鼠作为阴性对照组(NC组),另取各组大鼠颈椎间盘纤维环组织细胞进行原代细胞培养。应用HE染色和藏红O染色进行组织学分析。应用免疫组织化学染色、Western blot和RT-qPCR法检测MMP2、MMP9、金属蛋白酶组织抑制物2(TIMP2)和Ⅳ型胶原(collagenⅣ)的mRNA和蛋白表达。应用细胞-胶原黏附实验检测纤维环细胞-胶原黏附作用。结果:M组大鼠椎间盘出现退行性病变,黄芪多糖能够改善颈椎病大鼠椎间盘退行性病变。与NC组相比,M组大鼠纤维环组织中MMP2和MMP9表达水平显著增加,而TIMP2和collagenⅣ表达水平均显著降低(P 0. 05);与M组相比,L-AP组和H-AP组的MMP2和MMP9表达水平显著降低,而TIMP2和collagenⅣ的表达水平均显著增加(P 0. 05)。M组大鼠纤维环细胞-胶原黏附作用显著低于NC组(P 0. 05);与M组相比,L-AP组和H-AP组的纤维环细胞-胶原黏附作用均显著上升(P 0. 05)。与NC组相比,M组纤维环细胞中的MMP2和MMP9表达水平显著增加,而TIMP2和collagenⅣ的表达水平均显著降低(P 0. 05);与M组相比,L-AP组和H-AP组纤维环细胞中的MMP2和MMP9表达水平显著降低,而TIMP2和collagenⅣ表达水平均显著增加(P 0. 05)。结论:黄芪多糖能够抑制颈椎病模型大鼠纤维环组织中MMP2和MMP9表达,调节细胞外基质中MMPs与TIMPs的动态平衡,从而抑制MMPs对椎间盘基质中胶原的降解,在椎间盘退变的治疗中具有潜在研究价值。     

正常与退变椎间盘纤维环细胞相关基因的差异表达

目的: 研究人正常和退变椎间盘纤维环细胞椎间盘退变相关基因表达量的差异,探讨这些相关基因与椎间盘退变的关系,筛选明显差异基因作为退变纤维环细胞体外刺激因子,逆转椎间盘退变。方法: 实时荧光定量PCR(real-time qRT-PCR,SYBR Green)技术检测人正常和退变纤维环细胞TGF-β、IGF-1、bFGF、IL-1、TIMP-1、COL1A1、BMP-14、PDGF、aggrecan、COL2A1、SOX 9、BMP-2、iNOS、 EGF、 MMP3、BMP-4和BMP-7 mRNA表达水平。结果: 与正常纤维环细胞相比,退变纤维环细胞TGF-β、IL-1、PDGF和IGF-1 mRNA表达量降低(P<0.01,P<0.05);bFGF、TIMP-1、BMP-14和COL1A1 mRNA表达升量高(P<0.01);正常纤维环细胞aggrecan和BMP-2 mRNA高表达,退变纤维环细胞两者表达阴性;COL2A1、iNOS和MMP3在正常纤维环细胞mRNA有表达,在退变纤维环细胞表达阴性;EGF在正常纤维环细胞mRNA 表达阴性,而在退变纤维环细胞有表达。Sox-9、BMP-4和BMP-7在正常和退变纤维环细胞mRNA表达均为阴性。结论: 椎间盘纤维环退变与TGF-β、IL-1、PDGF、IGF-1、aggrecan和BMP-2 mRNA低表达,bFGF、TIMP-1、BMP-14和COL1A1 mRNA高表达相关;BMP-2、TGF-β、PDGF和IGF-1可作为退变纤维环细胞体外刺激因子,用于逆转椎间盘退变的研究;bFGF、TIMP-1和BMP-14 可能是椎间盘纤维环细胞退变的标志物。     

An annulus fibrosus closure device based on a biodegradable shape-memory polymer network

Injuries to the intervertebral disc caused by degeneration or trauma often lead to tearing of the annulus fibrosus (AF) and extrusion of the nucleus pulposus (NP). This can compress nerves and cause lower back pain. In this study, the characteristics of poly(d,l-lactide-co-trimethylene carbonate) networks with shape-memory properties have been evaluated in order to prepare biodegradable AF closure devices that can be implanted minimally invasively. Four different macromers with (d,l-lactide) to trimethylene carbonate (DLLA:TMC) molar ratios of 80:20, 70:30, 60:40 and 40:60 with terminal methacrylate groups and molecular weights of approximately 30 kg mol⁻¹ were used to prepare the networks by photo-crosslinking. The mechanical properties of the samples and their shape-memory properties were determined at temperatures of 0 °C and 40 °C by tensile tests- and cyclic, thermo-mechanical measurements. At 40 °C all networks showed rubber-like behavior and were flexible with elastic modulus values of 1.7–2.5 MPa, which is in the range of the modulus values of human annulus fibrosus tissue. The shape-memory characteristics of the networks were excellent with values of the shape-fixity and the shape-recovery ratio higher than 98 and 95%, respectively. The switching temperatures were between 10 and 39 °C. In vitro culture and qualitative immunocytochemistry of human annulus fibrosus cells on shape-memory films with DLLA:TMC molar ratios of 60:40 showed very good ability of the networks to support the adhesion and growth of human AF cells. When the polymer network films were coated by adsorption of fibronectin, cell attachment, cell spreading, and extracellular matrix production was further improved. Annulus fibrosus closure devices were prepared from these AF cell-compatible materials by photo-polymerizing the reactive precursors in a mold. Insertion of the multifunctional implant in the disc of a cadaveric canine spine showed that these shape-memory devices could be implanted through a small slit and to some extent deploy self-sufficiently within the disc cavity.     

体外培养人退变髓核与纤维环细胞增殖能力的研究

目的 探讨体外培养的人退变髓核与纤维环细胞的增殖能力,比较2种退变细胞体外培养中所表现的不同生物学行为,为椎间盘退变疾病的预防与治疗提供新的理论依据.方法 采集临床腰椎间盘突出症患者手术取材的椎间盘组织标本,病理学诊断评估其退变程度,酶消化法原代培养髓核与纤维环细胞,并鉴定.每例病例的髓核细胞与纤维环细胞分别体外培养至第5代,各代次细胞传代接种密度控制为1×105个.同条件培养48 h后,观察每一代细胞的形态学变化,同时应用流式细胞仪检测2种细胞的增殖能力.结果 体外培养的退变髓核与纤维环细胞生长状态良好.随着体外传代的进行,退变髓核与纤维环细胞S期细胞百分比和增殖指数(PI)均呈上升趋势,髓核细胞PI值于第3代时达到峰值,而纤维环细胞PI值于第5代时最高;且第2～4代的退变髓核细胞的增殖活性均高于退变纤维环的增殖活性(P＜0.05).结论 体外培养的人退变髓核与纤维环细胞有着不同的增殖特点,髓核与纤维环细胞对体内退变微环境有着不同的响应机制,并影响着整个椎间盘退变的进展.     

Hierarchical Structure of the Intervertebral Disc

Optical microscope techniques are used to characterize the hierarchical structure of the collagenous components of the human intervertebral disc. In the anterior annulus fibrosus, the thickness of lamellae increases abruptly 2 mm inward from the edge of the disc, dividing the annulus into peripheral and transitional regions. Lamellae in the lateral and posterior aspects of the disc have a broad distribution of lamellar thicknesses throughout the annulus. In alternating lamellae, fibers are inclined with respect to the vertical axis of the spine in a layup structure. From the edge of the disc inward to the nucleus, this interlamellar angle decreases from + 62 to + 45 degrees. Within lamellae, the collagen fibers exhibit a planar crimped morphology. The plane of the waveform is inclined with respect to the vertical axis by the interlamellar angle. From the edge of the disc inward, the crimp angle increases from 20 to 45 degrees and the crimp period decreases from 26 to 20 um. A hierarchical model of the intervertebral disc has been developed that incorporates these morphological gradients.     

Shape-memory porous alginate scaffolds for regeneration of the annulus fibrosus: Effect of TGF-β3 supplementation and oxygen culture conditions

Disc herniation as a result of degenerative or traumatic injury is believed to be the primary instigator of low back pain. At present there is a lack of viable treatment options to repair damaged annulus fibrosus (AF) tissue. Developing alternative strategies to fill and repair ruptured AF tissue is a key challenge. In this work we developed a porous alginate scaffold with shape-memory properties which can be delivered using minimally invasive approaches and recover its original geometry once hydrated. Covalently cross-linked alginate hydrogels were created using carbodiimide chemistry, followed by a freeze-drying step to impart porosity and create porous scaffolds. Results showed that porous alginate scaffolds exhibited shape-memory recovery and mechanical behaviour that could be modulated depending on the cross-linker concentrations. The scaffold can be repeatedly compressed and expanded, which provides the potential to deliver the biomaterial directly to the damaged area of the AF tissue. In vitro experiments demonstrated that scaffolds were cytocompatible and supported cell seeding, penetration and proliferation under intervertebral-disc-like microenvironmental conditions (low glucose media and low oxygen concentration). Extracellular matrix (ECM) was secreted by AF cells with TGF-β3 stimulation and after 21 days had filled the porous scaffold network. This biological matrix was rich in sulfated glycosaminoglycan and collagen type I, which are the main compounds of native AF tissue. Successful ECM deposition was also confirmed by the increase in the peak stress of the scaffold. However, the immaturity of the matrix network after only 21 days of in vitro culture was not sufficient to attain native AF tissue mechanical properties. The ability to deliver porous scaffolds using minimal invasive approaches that can potentially promote the regeneration of AF defects provides an exciting new avenue for disc repair.     

腰椎间盘的超微结构研究

本实验通过透射电镜对大鼠、家兔及人(胎儿及成人)腰椎间盘的观察,发现纤维环由纤维软骨组成。在640 A周期性的胶原原纤维之间,可见位于隐窝内的典型软骨细胞。这些细胞常随所在部位而异。靠近周边部位的细胞更接近成纤维细胞,细胞狭长,细胞膜的扇蛤样外形渐不明显,以至消失。靠内侧的细胞类似脊索细胞,细胞核固缩,细胞质内有浓集的糖原及脂滴。纤维环呈现细胞过渡的特点。纤维环内可见弹力纤维。髓核由包含细的原纤维、颗粒及特殊结构的疏松基质所组成。在各标本中,基质内均有脊索细胞出现。细胞可单独或成堆存在,细胞质内有众多的空泡,浓集的糖原颗粒及微丝。     

循环牵张应力对人退变纤维环细胞增殖的影响

目的 探讨不同水平循环牵张应力对人退变椎间盘细胞增殖的影响.方法 取1例29岁手术切除的腰椎间盘突出患者的椎间盘,通过病理学方法评价其退变程度,酶消化法分离椎间盘纤维环细胞.选取P3代细胞,利用ElectroForce 3200力学试验仪搭载的BioDynamic生物反应舱系统,以3h为固定时间点对其进行不同水平的频率...     

仿生可降解组织工程纤维环支架的制备与评估

文题释义： 纤维环：是保持椎间盘强度与脊柱稳定性层面的重要组成部分,纤维环的胶原纤维在椎间盘内呈同心圆排列,每层纤维环间呈60°夹角斜行排列,这类特殊的交错网状架构,令纤维环拥有较强的抗拉性能与压缩性,能够预防髓核往外突出,对保持椎间盘的稳定起着重要的作用。 背景：构建既具有仿生结构又具备合适可降解性和良好生物相容性的组织工程纤维环支架仍是难点。 目的：以聚己内酯和聚二恶烷酮为原材料制备仿生可降解支架并评估其作为组织工程纤维环支架的可行性。 方法：通过熔融纺丝技术制备5组不同比例支架(聚己内酯组、聚己内酯/聚二恶烷酮分别为70/30、50/50、30/70组、聚二恶烷酮组)。扫描电子显微镜观察其结构、纤维直径、孔径;测量支架的力学性能和接触角;通过体外模拟和皮下埋植观察支架体外、体内降解情况;检测降解组织周围炎症因子白细胞介素1β和肿瘤坏死因子α的表达情况。接种人脐带间充质干细胞培养7 d,通过CCK-8和死活细胞检测细胞增殖和存活情况。实验于2016-03-02经天津市天津医院医学伦理委员会批准。 结果与结论：①扫描电子显微镜观察显示各组支架纤维粗细均匀,纤维成60°角;②力学性能分析显示单纯聚二恶烷酮支架的拉伸和压缩模量最低,不符合纤维环力学要求;聚己内酯组的力学性能最佳,聚己内酯/聚二恶烷酮为70/30和50/50的支架力学性能适中;③亲水性检测结果表明聚二恶烷酮含量越多,支架亲水性越好;④支架降解情况分析显示,对于组织工程纤维环再生修复来说,单纯聚二恶烷酮和聚己内酯/聚二恶烷酮为30/70支架降解过快,聚己内酯组的降解过慢,聚己内酯/聚二恶烷酮为70/30和50/50的支架降解速率较合适;⑤降解组织周围炎症反应分析显示支架中聚己内酯比例越高炎症反应越严重;⑥CCK-8和死活细胞结果显示人脐带间充质干细胞在聚己内酯/聚二恶烷酮三组混合支架具有良好的增殖活性并且存活率高;⑦结果表明,采用熔融纺丝技术制备的聚己内酯/聚二恶烷酮为70/30和50/50两组支架能够模拟天然纤维环结构,同时具备合适可降解性、优越的力学性能和良好生物相容性,适合用于组织工程纤维环支架的构建。 ORCID: 0000-0001-9088-4222(张维昊) 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

芍药苷促椎间盘纤维环细胞增殖的作用

背景：芍药在颈椎病治疗中使用频率超过30%,其主要单体之一芍药苷具有抗炎、抗凋亡等作用,芍药苷对椎间盘纤维环细胞是否有保护作用,国内外未见报道。 目的：探讨芍药苷对椎间盘纤维环细胞的增殖和保护作用。 方法：采用酶消化法体外培养椎间盘纤维环细胞。取第3代纤维环细胞,培养过程中加入20 μg/L Fas配体诱导建立纤维环细胞凋亡模型,通过MTT法测定芍药苷干预纤维环细胞的最适宜浓度,Annexin V/FITC-PI法观察芍药苷对椎间盘纤维环细胞的保护作用。 结果与结论：芍药苷浓度20.8和2.08 μmol/L对椎间盘纤维环细胞具有明显的促增殖作用。20 μg/L Fas配体能诱导细胞凋亡,20.8 μmol/L芍药苷对Fas配体诱导椎间盘纤维环细胞有保护作用。     

腰椎4、5间盘纤维环纤维层力学性能的实验研究

测试成人腰椎间盘纤维环纤维层的力学性能。用显微外科技术将纤维环逐层分离,每层纤维平均分成8个测点,每层每测点处按纤维走向切取5个试件,长度l=15~20mm,宽b=1~1.5mm,厚t=0.1~0.5mm。进行拉伸实验测得应力-应变曲线,并给出拟合曲线及方程,得出弹性模量、破坏应变、破坏应力、测点处的力学性能参数和方程。测试结果表明:各测点处,沿径向,由外向内弹性模量皆随层数的增加,不断变小;每层纤维弹性模量由前侧向后侧逐渐变小;破坏应力由外层向内层线性递减;破坏应变外部9层随层数增加略有提高,9层以后基本上为一常数(0.34±0.14)。可见,腰椎间盘纤维环纤维具有特殊的力学性能,与其功能相适应,与腰椎疾病的发生有密切的关系。     

腰4、5间盘纤维环结构形态的细微观察

观测成人腰椎间盘纤维环纤维层水平的细微解剖结构。用显微外科技术将腰间盘纤维环分离,每层纤维均分为8个观测点,测量纤维走向与水平面的夹角等数值。结果表明:各测点处的纤维走向角α,在水平面,沿径向由外向内,随层数的增加不断变大,最小纤维走向角发生在前正中处,其值为25°~30°;在同一层,纤维走向角由前向后不断变大,在椎间盘正后方为70°~90°,即此部分纤维走向与后纵韧带走向基本一致;用归一化方程,归一化直线可以非常方便地求出任一层、任一点处的纤维走向角α;腰椎间盘纤维环的前方、左右两侧皆有交织现象发生,左右两侧交织区域多些,仅后方正中附近未发现相临两层的交织现象。由此可见,腰椎间盘纤维环纤维具有特殊的细微结构,与其功能相适应。     

Immunolocalization of major interstitial collagen types in human lumbar intervertebral discs of various ages

 We used complete transverse sections through 65 samples of human lumbar intervertebral discs for immunolocalization of the major interstitial collagen types I, II, III, V, VI and IX. The samples were selected from 47 patients ranging in age from 0 (fetuses) to 86 years. The results were compared with the histological findings in disc tissue degeneration and/or reparative alterations as indicated by tear and cleft formation, chondrocyte proliferation, mucous degeneration, granular matrix changes and fibrocartilage fibrillation. We observed a typical pattern for each antibody and each anatomical structure, with, however, remarkable inter- and intraindividual variability, which could be monitored only by use of the complete transverse sections. Accordingly, collagen I was seen in the normal annulus fibrosus and in the degeneratively altered nucleus pulposus, but not within the end-plate, regardless of degenerative changes. Collagens II and IX were found in the normal nucleus pulposus, the inner annulus fibrosus and the end-plate. The collagen II (and IX) staining seemed to be enhanced in areas of minor degenerative lesions, but reduced in advanced lesions and in the degenerated end-plate. Collagens III and VI were significantly increased in areas of minor to advanced degeneration in all anatomical settings, while collagen V showed only minor changes in its staining pattern. In general, histological signs of tissue degeneration coincided with significant quantitative, but also with certain qualitative, changes in the composition of the collagenous disc matrix. These observations indicate the association of degenerative and/or reparative alterations of the intervertebral disc and changes in the collagenous matrix, but document the variability in the extent of the abnormalities observed. Received: 5 December 1996 / Accepted: 30 May 1997     

Anisotropic shear behavior of the annulus fibrosus: effect of harvest site and tissue prestrain

We used a shear protocol to investigate the time-independent, anisotropic structural behavior of 30 cube and 56 sheet specimens of human annulus fibrosus from six non-degenerate lumbar spines. The specimen sides were aligned with the natural collagen lamellar architecture. For the cube specimens, simple shear was applied sequentially in three orthogonal directions defined by the cube axes. The sheets were also tested in simple shear, with the stress applied in either the anatomic axial or circumferential directions. With the sheet specimens we also investigated the contribution of annular collagen to the shear modulus by applying a tensile prestrain (0, 5 or 10 percent) perpendicular to the direction of applied shear stress. Our cube data indicated that the shear modulus was anisotropic, being 56.04+/-36.3 kPa in the plane of the lamellae and approximately half that in the orthogonal directions. The sheet specimens demonstrated that shear modulus increased progressively by a factor of between 3 and 5 from the inner to outer annulus. The ratio of the axial to circumferential shear modulus for the sheets increased from being near unity for the inner annulus to near 2 for the outer annulus. Finally, the addition of a 10 percent tensile prestrain increased the shear modulus by between 1.5 and 2.5 times for the middle and outer annulus. The shear anisotropy we report is consistent with prior anatomical observations of this tissue and appears to develop through separate contributions from the matrix, the collagen fibers, and collagen fiber interactions.     

Diffusivity of Ions in Agarose Gels and Intervertebral Disc: Effect of Porosity

The effect of tissue porosity on ion (sodium, potassium, and chloride) diffusivity in agarose gels and porcine intervertebral disc tissues was investigated using an electrical conductivity method. An empirical, constitutive model for diffusivity (D) of solutes in porous fibrous media was proposed: D/D_o=exp[–(r_s/^1/2)] where r_s is the Stokes radius of a solute, is the Darcy permeability of the porous medium, D_o is the diffusivity in free solution, and are two positive parameters whose values depend on material structure. It is found that =1.25±0.138, =0.681±0.059 (95% confidence interval, R²=0.92, n=72) for agarose gels and =1.29±0.171 and =0.372±0.088 (95% confidence interval, R²=0.88, n=86) for porcine annulus fibrosus. The functional relationship between solute diffusivity and tissue deformation was derived. Comparisons of our model prediction with experimental data on diffusion coefficients of macromolecules (proteins, dextrans, polymer beads) in agarose gels in the literature were made. Our results were also compared to the data on ion diffusivity in charged gels and in cartilaginous tissues reported in the literature. There was a good agreement between our model prediction and the data in the literature. The present study provides additional information on solute diffusivity in uncharged gels and charged tissues, and is important for understanding nutritional transport in avascular cartilaginous tissues under different mechanical loading conditions.     

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。     

外科生物补片修复退变椎间盘纤维环

BACKGROUND: In recent years, animal models of lumbar disc degeneration have been popularized to explore the effect of tissue engineering, cell engineering and genetic engineering technologies on intervertebral disc degeneration.     

Regional variations in the density and arrangement of elastic fibres in the anulus fibrosus of the human lumbar disc

Elastic fibres are critical components of the extracellular matrix in dynamic biological structures that undergo extension and recoil. Their presence has been demonstrated in the anulus fibrosus of the human lumbar intervertebral disc; however, a detailed regional analysis of their density and arrangement has not been undertaken, limiting our understanding of their structural and functional roles. In this investigation we have quantitatively described regional variations in elastic fibre density in the anulus fibrosus of the human L3-L4 intervertebral disc using histochemistry and light microscopy. Additionally, a multiplanar comparison of patterns of elastic fibre distribution in the intralamellar and interlamellar zones was undertaken. Novel imaging techniques were developed to facilitate the visualization of elastic fibres otherwise masked by dense surrounding matrix. Elastic fibre density was found to be significantly higher in the lamellae of the posterolateral region of the anulus than the anterolateral, and significantly higher in the outer regions than the inner, suggesting that elastic fibre density in each region of the anulus is commensurate with the magnitude of the tensile deformations experienced in bending and torsion. Elastic fibre arrangments in intralamellar and interlamellar zones were shown to be architecturally distinct, suggesting that they perform multiple functional roles within the anulus matrix structural hierarchy.