华西口腔医学杂志

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自组装山羊颞下颌关节盘组织工程纤维软骨模型的构建

康宏 李振强 闭艳妲   

  1. 兰州大学口腔医学研究所, 兰州730000
  • 收稿日期:2011-06-25 修回日期:2011-06-25 出版日期:2011-06-20 发布日期:2011-06-20
  • 通讯作者: 康宏,Tel:0931-6112809
  • 作者简介:康宏(1966—),男,甘肃人,教授,博士
  • 基金资助:

    甘肃省国际科技合作基金资助项目(0804WCGA127)

Self-assembly tissue engineering fibrocartilage model of goat temporomandibular joint disc

KANG Hong, LI Zhen-qiang, BI Yan-da   

  1. Institute of Stomatology, Lanzhou University, Lanzhou 730000, China
  • Received:2011-06-25 Revised:2011-06-25 Online:2011-06-20 Published:2011-06-20
  • Contact: KANG Hong,Tel:0931-6112809

摘要:

目的构建山羊颞下颌关节盘纤维软骨自组装模型,观察自组装组织工程纤维软骨的生物学特征,为颞下颌关节盘及其他组织工程纤维软骨的进一步研究创造条件。方法分离、培养山羊颞下颌关节盘细胞,按每井5.5×106个接种到预制的直径5 mm×深10 mm琼脂糖井内,每天换液,培养2周,观察和检测颞下颌关节盘形态和成分的变化。结果接种后1 d,山羊颞下颌关节盘细胞在琼脂糖井内聚集,开始自组装成一圆盘状基体,后来逐渐变成圆球形。培养2周时,苏木精-伊红染色可观察到纤维软骨细胞呈圆形,周围有基质包绕;Safranin-O/fast green染色显示自组装基体内纤维软骨细胞分泌细胞外基质氨基多糖;picro-sirius red染色可见自组装基体内有大量胶原纤维。Ⅰ型胶原免疫组织化学染色可见胞浆和细胞外有棕黄色颗粒,表明自组装纤维软骨可以产生与自然关节盘组织一致的Ⅰ型胶原成分。结论利用琼脂糖井构建的山羊颞下颌关节盘自组装组织工程模型,能产生与自然关节盘成分相似的细胞外基质,表明组织工程化颞下颌关节盘组织的途径是可行的。

关键词: 颞下颌关节盘, 组织工程, 纤维软骨

Abstract:

Objective To construct self-assembly fibrocartilage model of goat temporomandibular joint disc and observe the biological characteristics of the self-assembled fibrocartilage constructs, further to provide a basis for tissue engineering of the temporomandibular joint disc and other fibrocartilage. Methods Cells from temporomandibular joint discs of goats were harvested and cultured. 5.5×106 cells were seeded in each agarose well with diameter 5 mm × depth 10 mm, daily replace of medium, cultured for 2 weeks. Results One day after seeding, goat temporomandibular joint disc cells in agarose wells were gathered and began to self-assemble into a disc-shaped base, then gradually turned into a round shape. When cultured for 2 weeks, hematoxylin-eosin staining was conducted and observed that cells were round and wrapped around by the matrix. Positive Safranin-O/fast green staining for glycosaminoglycans was observed throughout the entire constructs, and picro-sirius red staining was examined and distribution of numerous type Ⅰ collagen was found. Immunohistochemistry staining demonstrated brown yellow particles in cytoplasm and around extracellular matrix, which showed self-assembly construct can produce type Ⅰ collagen as native temporomandibular joint disc tissue. Conclusion Production of extracellular matrix in self-assembly construct as native temporomandibular joint disc tissue indicates that the use of agarose wells to construct engineered temporomandibular joint disc will be possible and practicable.

Key words: temporomandibular joint disc, tissue engineering, fibrocartilage