Xu Hong-guang, Peng Hong-xin, Cheng Jia-feng, Lü Kun
Department of Orthopedic Surgery, Wannan Medical College, Wuhu, China.
Zhonghua Yi Xue Za Zhi. 2011 Nov 8;91(41):2912-6.
To establish an in vitro model of degeneration of human cervical endplate chondrocytes and observe the morphology and phenotypes of endplate chondrocytes in normal and degenerative cervical vertebral endplates.
Cartilage endplates of 49 patients were divided into control group (n = 19) with cervical vertebral fracture or dislocation and experiment group (n = 30) with cervical spondylotic myelopathy. Endplate chondrocytes were isolated by enzyme digestion and cultured in vitro. The morphological appearances, growth curve and biological characteristics of endplate chondrocytes from normal and degenerative cartilage endplate were observed by inverted phase contrast microscope, HE staining, MTT, toluidine blue staining and reverse transcription-polymerase chain reaction (RT-PCR) respectively. RT-PCR was used to detect the mRNA expression of aggrecan, type II collagen and type I collagen.
The endplate chondrocytes expressed aggrecan, type II collagen and type I collagen. The phenotypes and biological characteristics were similar to those of articular chondrocytes. The morphological appearance of primary endplate chondrocytes in the control group were mostly polygons, nucleus with round or ellipse, sometimes nuclei, vacuoles in intra cytoplasm, expressing a high proliferating rate. The cells of the experiment group were fusiform and their proliferating rates decreased. Compared with the control group, the mRNA expression of aggrecan (0.695 ± 0.052 vs 0.950 ± 0.032, t = 7.263, P = 0.002) and type II collagen (0.726 ± 0.035, 0.907 ± 0.078, t = 3.681, P = 0.021) markedly decreased. And the mRNA expression of type I collagen (0.795 ± 0.028 vs 0.552 ± 0.070, t = -5.560, P = 0.005) increased in the experiment group.
A degenerative cell model of human cervical endplate chondrocytes has been established successfully in vitro. It may offer the cytological rationales for exploring the mechanism of intervertebral disc degeneration. And the previous restrictions of studying only the model of animal cells shall be resolved.
建立人颈椎终板软骨细胞退变的体外模型，观察正常及退变颈椎终板中终板软骨细胞的形态和表型。
将49例患者的软骨终板分为对照组（n = 19），包括颈椎骨折或脱位患者；实验组（n = 30），包括脊髓型颈椎病患者。采用酶消化法分离终板软骨细胞并进行体外培养。分别通过倒置相差显微镜、苏木精-伊红染色、MTT法、甲苯胺蓝染色及逆转录-聚合酶链反应（RT-PCR）观察正常及退变软骨终板来源的终板软骨细胞的形态、生长曲线及生物学特性。RT-PCR检测聚集蛋白聚糖、Ⅱ型胶原和Ⅰ型胶原的mRNA表达。
终板软骨细胞表达聚集蛋白聚糖、Ⅱ型胶原和Ⅰ型胶原。其表型和生物学特性与关节软骨细胞相似。对照组原代终板软骨细胞形态多为多边形，细胞核呈圆形或椭圆形，有时可见双核及胞质内空泡，增殖率较高。实验组细胞呈梭形，增殖率降低。与对照组比较，实验组聚集蛋白聚糖（0.695±0.052比0.950±0.032，t = 7.263，P = 0.002）和Ⅱ型胶原（0.726±0.035比0.907±0.078，t = 3.681，P = 0.021）的mRNA表达明显降低。实验组Ⅰ型胶原的mRNA表达升高（0.795±0.028比0.552±0.070，t = -⁵.⁵⁶⁰，P = 0.005）。
成功建立了人颈椎终板软骨细胞退变的细胞模型。可为探讨椎间盘退变机制提供细胞学依据，解决以往仅研究动物细胞模型的局限性。
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