(解放军第306医院耳鼻咽喉头颈外科,),北京,100101
网络首发:2017-01-20,
纸质出版:2017
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周丽斌, 徐冰心, 丁瑞英, 等. 应用纤维蛋白胶复合细胞膜片构建人工软骨[J]. 中山大学学报(医学科学版), 2017,38(1).
Cartilage Engineering Using Fibrin Gel and Chondrocyte Cell Sheets[J]. Journal of Sun Yat-sen University (Medical Sciences), 2017, 38(1).
摘要:【目的】利用细胞膜片具备的良好的流动性,以及纤维蛋白胶的可塑性为其塑形,构建具备空间立体结构的软 骨块。【方法】采用兔耳软骨细胞在培养瓶中进行体培养形成细胞膜片,刮取后剪碎,另取离散软骨细胞,用纤维蛋白胶两种 组分混悬细胞膜片和离散软骨细胞,滴加到48孔板中,在体外构建出圆片状的结构,经过体外培养后进一步植入裸鼠体内,培 养8周后取材。采用大体观察及组织学、免疫组织化学等方法观察分析体内外培养的结构。【结果】获得成熟的圆片状组织工 程软骨结构,组织学表现为成熟的软骨组织,免疫组化结果提示软骨细胞外基质内II型胶原蛋白高表达。但结构内软骨组织分 布不均匀,岛状软骨组织之间的间隙内为未降解的纤维蛋白胶成分。在其中1个圆片结构中,发现局部骨化现象。【结论】纤维 蛋白胶是一种良好的载体,用于复合软骨细胞膜片及软骨细胞,可成功构建出具备空间立体结构人工软骨结构。
Abstract:【Objective】To study the feasibility of Cartilage engineering using fibrin gel and chondrocyte cell sheets.【Methods】 rabbit auricular chondrocytes were isolated and cultured to form cell sheets in flasks. The cell sheets were harvested using cell scrapers,and cut into fragments. The two precursor solutions of Fibrin gel were used to suspend the cell sheet fragments and isolated chondrocytes,and then added into the wells of a 48-well plate to form Gelatinous chondroid disc constructs. After in vitro culture, the constructs were implanted into nude mice. After 8 weeks,the constructs were harvested,and the specimens were evaluated using grossly observing, histological and immunohistochemical observation. 【Results】Mature cartilage discs were obtained. The histomorphology of the explanted discs appeared non- uniform cartilaginous tissue comprise of regenerated cartilage islands with different size and irregular shape. Immunohistochemistry staining demonstrated that type II collagen highly expressed in the ECM of the cartilage islands. In 1 of the 8 discs,partial ossification was observed.【Conclusion】Fibrin gel is a favourable carrier. Artificial cartilage with stereochemical structure was constructed via combining the fibrin gel and chondrocyte cell sheets.
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