Mallory染色法在椎间盘退变组织染色中的应用

Application of Mallory trichrome staining in degenerative tissues of intervertebral disc

背景:椎间盘退变的同时伴随有组织结构和成分的改变。采用多种染色剂连续复染形成多元色的组织学图形,可观察到各组织成分的不同颜色以及椎间盘退变时的颜色变化,在分辨力的显现上,比单纯形态学的改变更明显且易于分辨。目的:从组织形态和颜色变化两方面观察Mallory三色染色对椎间盘的染色效果。方法:取1,1.5,2,2.5和3月龄Hartley豚鼠L4～5椎体,制备中央冠状面椎间盘组织切片。用Mallory染液染色,光学显微镜下观察髓核、软骨终板和纤维环各区域形态和颜色的变化。SPOT-Ⅱ数码成像系统拍摄图像。结果与结论:Hartley豚鼠椎间盘退变程度随鼠龄增长自然加重。1月龄豚鼠的髓核形态正常,无色透明。在1.5～3月龄豚鼠中,髓核逐渐退变,由局部发展至全部基质呈现浅蓝着色,表明有胶原沉积,其形态上可见脊索细胞呈现软骨样细胞变性,髓核面积逐渐缩小并逐渐被纤维软骨样组织替代,直至在3月龄时呈现"蛇纹石"样外观。其终板上紫红色软骨细胞带随鼠龄增长呈现出不规则形态直至消失,其形态上可见软骨细胞逐渐减少。外纤维环红色和橘黄色的面积随着鼠龄增长而增加,表明有纤维素样变性,其形态上可见明显的板层状结构。内纤维环蓝色的面积随着鼠龄增长逐渐缩小。结果显示Mallory三色法用于Hartley豚鼠椎间盘染色时,在髓核、软骨终板和纤维环上,可从色彩、形态两方面显示出对基质和细胞具有良好的组织分辨力,能从形态和成分变化的角度反映自然增龄过程中椎间盘组织的退变情况。

BACKGROUND：Intervertebral disc degeneration accompanies by alterations in tissue structure and component.The component changes of intervertebral disc degeneration can be displayed in different colors when applying various stains for successive counterstaining.This method exhibits detectable changes than simple morphological observation.OBJECTIVE：To observe the staining effect of the intervertebral disc by Mallory trichrome staining from the changes of both tissue morphology and colors.METHODS：L4-5 intervertebral disc of Hartley guinea pigs,which aged 1,1.5,2,2.5 and 3 months,respectively.Discs were made into the central cornal plane tissue sections.Mallory staining and a light microscope was used to observe morphology and component and colors changes of nucleus pulposus,cartilage endplate and annulus fibrosus.SPOT-Ⅱ digital imaging system was utilized to capture images.RESULTS AND CONCLUSION：Disc degeneration of Hartley guinea pigs aggravated naturally with age.Normal（1 month old） nucleus pulposus was colorless and transparent.At 1.5-3 months old,matrix of nucleus showed light blue color from local to all when degeneration,to indicate collagen eposition.Morphologically,notochord cells showed the degeneration of cartilage-like cells,the nucleus area was gradually narrowing and replaced by fibrous cartilage tissue.Nucleus was like to ＂serpentine＂ in modality at 3 months old.In the cartilage endplate,the purplish red cartilage cells belt showed irregular until disappear with age increasing.Morphologically,the cartilage cells were decreasing gradually,and the endplate was thinning,broken or disappeared.Red and orange area of outer annulus increased with age,to indicate fibrinoid degeneration.Lamellar structure could be seen clearly.The blue area of inner annulus was reduced gradually with age and the number of fibroblasts was decreased and disappeared.The results display that Mallory trichrome staining for Hartley guinea pigs intervertebral disc can show a good organizational resolutionin from both color and morphology,for matrix and cell which in the nucleus pulposus,cartilage endplate and the annulus,and visualizing the characteristic changes of intervertebral disc in the process of the natural increase of age from both morphology and component change.