A Stereological Method for the Quantitative Evaluation of Cartilage Repair Tissue
A Stereological Method for the Quantitative Evaluation of Cartilage Repair Tissue
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DOI：
10.1177/1947603514560655
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发表时间：
2015-04-01
期刊：
影响因子：
2.8
通讯作者：
Spector, Myron
中科院分区：
文献类型：
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作者：
Foldager, Casper Bindzus;Nyengaard, Jens Randel;Spector, Myron
目的：应用体视学原理，建立一种简便易行的无偏定量评价软骨修复的算法。设计图：通过在平行平面中垂直于关节表面对缺损进行系统切片，提供7 - 10个苏木精-伊红染色的组织学切片，进行设计无偏倚取样。系统地选择计数窗口并将其转换为图像文件（每个缺陷40-50个）。通过两步点计数进行定量：（1）计算缺损体积和（2）定量分析组织成分。通过基于经验证且易于区分的形态学特征将每个点分配到以下类别之一来执行步骤2：（1）透明软骨（透明基质中陷窝中的圆形细胞），（2）纤维软骨（纤维基质中陷窝中的圆形细胞），（3）纤维组织（纤维组织中的细长细胞），（4）骨，（5）支架材料和（6）其他。使用常规或偏振光显微镜确定区分组织类型的能力，并评价观察者间的变异性。结果：描述了体视学方法的应用。在本例中，我们评估缺损修复组织体积为4.4 mm（3）（CE = 0.01）。随后评价组织部分。偏振光照明的幻灯片提高了透明软骨和纤维软骨之间的歧视，并增加了观察者之间的协议相比，传统的透射光。结论：我们已经应用了一个设计无偏的方法定量评价软骨修复，我们提出了这种算法作为一个自然的补充，现有的描述性半定量评分系统。我们还建议偏振光是有效的区分透明软骨和纤维软骨。
Objective: To implement stereological principles to develop an easy applicable algorithm for unbiased and quantitative evaluation of cartilage repair. Design: Design-unbiased sampling was performed by systematically sectioning the defect perpendicular to the joint surface in parallel planes providing 7 to 10 hematoxylin-eosin stained histological sections. Counting windows were systematically selected and converted into image files (40-50 per defect). The quantification was performed by two-step point counting: (1) calculation of defect volume and (2) quantitative analysis of tissue composition. Step 2 was performed by assigning each point to one of the following categories based on validated and easy distinguishable morphological characteristics: (1) hyaline cartilage (rounded cells in lacunae in hyaline matrix), (2) fibrocartilage (rounded cells in lacunae in fibrous matrix), (3) fibrous tissue (elongated cells in fibrous tissue), (4) bone, (5) scaffold material, and (6) others. The ability to discriminate between the tissue types was determined using conventional or polarized light microscopy, and the interobserver variability was evaluated. Results: We describe the application of the stereological method. In the example, we assessed the defect repair tissue volume to be 4.4 mm(3) (CE = 0.01). The tissue fractions were subsequently evaluated. Polarized light illumination of the slides improved discrimination between hyaline cartilage and fibrocartilage and increased the interobserver agreement compared with conventional transmitted light. Conclusion: We have applied a design-unbiased method for quantitative evaluation of cartilage repair, and we propose this algorithm as a natural supplement to existing descriptive semiquantitative scoring systems. We also propose that polarized light is effective for discrimination between hyaline cartilage and fibrocartilage.