自组装山羊颞下颌关节盘组织工程纤维软骨模型的构建

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

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。

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 through-out 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.Con-clusion Production of extracellular matrix in self-assembly construct as native temporomandibular joint disc tissue indi-cates that the use of agarose wells to construct engineered temporomandibular joint disc will be possible and practicable.