生物蛋白胶/松质骨基质负载软骨细胞体外培养的实验研究

Biological fibrin glue/cancellous bone matrix engineered chondrocyte cultured in vitro

目的探讨生物蛋白胶/松质骨基质作为软骨细胞培养支架的可行性。方法①取3周龄的新西兰幼兔的关节软骨细胞种植于生物蛋白胶/松质骨基质载体上作体外培养;②通过倒置显微镜、扫描电镜、透射电镜、AB-PAS染色、Ⅱ型胶原免疫组化检查等,了解细胞生长及软骨基质的分泌情况,并观察载体降解情况。结果①生物蛋白胶/松质骨基质载体中大部分保持卵圆形,小部分在支架表面如贴壁细胞状;②于第9天以后,支架内的细胞数均开始明显增加。软骨细胞活性达95%以上;③AB-PAS染色(+)、Ⅱ型胶原免疫组化检查(+);④电镜观察显示软骨细胞较好地附在支架上并分泌细胞外基质。结论生物蛋白胶/松质骨基质是一种较好的软骨细胞培养支架。

Objective To investigate the feasibility of biological fibrin glue/cancellous bone matrix as scaffold for chondrocyte. Methods ①Chondroeytes taken from New Zealand rabbits aged 3 weeks were seeded onto biological fibrin glue/cancellous bone matrix and eultm-ed in vitro. ②Phase contract microscopy, electron microscopy, AB-PAS stain and inmunohistochemisting of collagen Ⅱ were used to study the behavior of ehondrocytes cultured in the scaffolds. Results ①Most ehondrocytes retained their oval shape when cultured in biological fibrin glue/cancellous bone matrix. ②The number of ehondrocytes in the scaffolds increased significantly after 7 days, and the survival rate of ehondrocytes was 95% .③AB-PAS stain was（ ＋ ）,type 11 collagen inmmtmohistocbemieal test （ ＋ ）. Electronic microscopic examination revealed that chondroeytes grow well and could .synthesize cartilage extracellular matrix in the scaffold. Conelusion Biological fibrin glue/cancellous bone matrix could be a good scaffold for ehondrocyte culture.