应用纤维蛋白胶复合细胞膜片构建人工软骨

【Objective】To utilize the good fluidity of cell sheets and the plasticity of fibrin glue to shape them and construct a cartilage block with a three-dimensional spatial structure. 【Methods】Rabbit auricular chondrocytes were cultured in vitro in a culture flask to form cell sheets, which were scraped and cut into pieces. Additionally, dispersed chondrocytes were obtained. The two components of fibrin glue were used to suspend the cell sheets and dispersed chondrocytes, and the mixture was dropped into a 48-well plate to construct a circular sheet-like structure in vitro. After in vitro culture, it was further implanted into nude mice, and samples were taken 8 weeks after culture. Gross observation, histology, immunohistochemistry and other methods were used to observe and analyze the structures cultured in vivo and in vitro. 【Results】A mature circular sheet-like tissue-engineered cartilage structure was obtained. Histologically, it showed a mature cartilage tissue, and the immunohistochemical results indicated a high expression of type II collagen in the extracellular matrix of chondrocytes. However, the distribution of cartilage tissue within the structure was not uniform, and the gaps between the island-like cartilage tissues contained undegraded fibrin glue components. In one of the circular sheet structures, local ossification was found. 【Conclusion】Fibrin glue is a good carrier for compounding cartilage cell sheets and chondrocytes, and an artificial cartilage structure with a three-dimensional spatial structure can be successfully constructed.

【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。