组织工程化软骨修复界面整合的体外模型

背景：随着组织工程学的发展，自体软骨细胞移植技术经常被用来修复软骨缺损，整合不良是导致修复失败的原因之一。许多体外模型被用来进行这方面的研究。 目的：建立一种组织工程化软骨修复界面整合的体外实验模型并评价其效果。 方法：制备猪体外软骨整合模型，获得21个软骨环，18只琼脂糖凝胶覆盖的软骨环设为琼脂糖凝胶组，剩余3个做无琼脂糖对照组，分别植入分离的软骨细胞，观察近期软骨环边界细胞漏出情况，分别在1，2，4周做切片、染色并行组织学观察，测量新生软骨平均面积并进行比较。 结果与结论：无琼脂糖对照组由于软骨细胞早期从软骨环底部漏出，未能在软骨环中形成软骨细胞聚集，所以未做后期处理，而琼脂糖凝胶组则未发生。琼脂糖凝胶组1，2，4周做切片并行固定后组织切片分别用苏木精-伊红染色、阿利新蓝、番红O、Ⅱ型胶原免疫组化染色，移植的软骨细胞在软骨环内不断增殖，并且产生细胞外基质。在第1，2周的孵育中，新生软骨的面积明显增大，到第4周时，面积也有进一步增加，但是第2-4周的面积增加，差异无显著性意义（P〉0.05）。模型成功模拟了自体软骨细胞移植修复关节软骨缺损的体外整合过程，未来可应用于软骨整合及软骨组织工程的机制研究。

An integrated model for tissue engineered cartilage repair in vitro

BACKGROUND：With the development of tissue engineering, autologous chondrocyte implantation is often used to repair cartilage defects. And poor integration is one of the common reasons that lead to failure repairing. Many models in vitro are used for related studies. OBJECTIVE：To develop an interface integrated model of tissue engineered cartilage repair in vitro and to evaluate the effect. METHODS：Cartilage integration model in vitro was established in pigs. Total y 21 cartilaginous rings were obtained and divided into agarose gel group （n=18） and control group （n=3）. In agarose gel group, cartilage rings were covered with agarose gel. Chondrocytes were separated and implanted into the ring. The leakage of cells around the cartilage rings was observed. The sections were stained for histological observation at 1, 2, 4 weeks. The average area of neochondrocytes was measured and compared. RESULTS AND CONCLUSION：The results from the control group were not processed, because there was no chondrocyte aggregate formation in the center of the explant ring due to earlier chondrocyte leakage outside the explant. While no chondrocytes were found outside the explant ring in the agarose gel group. Tissue sections of the agarose gel group were stained by hematoxylin and eosin, alcian blue, Safranin-O and col agen type II immunohistochemistry at 1, 2, 4 weeks. Neochondrocytes proliferated within cartilage ring, and produced＆amp;nbsp;extracellular matrix. After 2 weeks of incubation, these inserted chondrocytes were significantly increased. There was no statistical y significant increment between 2 weeks and 4 weeks （P〉0.05）, although the area was further increased by 4 weeks. This model provides a convenient simulation of the cartilage integration process in vitro and has a potential application in studies of cartilage integration and cartilage tissue engineering.