组织工程化软骨修复界面整合的体外模型.doc

组织工程化软骨修复界面整合的体外模型周建新1，高 峰2，桂鉴超1，尹昭伟1，杨晓斐1，徐 扬1，陆一鸣1，李 杨1，蒋逸秋11南京医科大学附属南京医院骨科，南京市第一医院，江苏省南京市 210006；2盐城市第三人民医院，江苏省盐城市 224000An integrated model for tissue engineered cartilage repair in vitroZhou Jian-xin1, Gao Feng2, Gui Jian-chao1, Yin Zhao-wei1, Yang Xiao-fei1, Xu Yang1, Lu Yi-ming1, Li Yang1, Jiang Yi-qiu11 Department of Orthopedics, Nanjing Hospital Affiliated to Nanjing Medical University, Nanjing First Hospital, Nanjing 210006, Jiangsu Province, China; 2 Third Peoples Hospital of Yancheng, Yancheng 224000, Jiangsu Province, China摘要 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、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。中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程全文链接：窗体顶端关键词 ： 实验动物； 组织构建； 软骨修复； 自体软骨植入； 体外模型； 整合； 细胞移植； Abstract： 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. Totally 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 collagen type II immunohistochemistry at 1, 2, 4 weeks. Neochondrocytes proliferated within cartilage ring, and produced extracellular matrix. After 2 weeks of incubation, these inserted chondrocytes were significantly increased. There was no statistically 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.中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程全文链接：Key words： tissue engineering; cartilage; chondrocytes; cell transplantation; cartilage, articular; 窗体底端 中图分类号:R318 通讯作者: 桂鉴超，硕士生导师，主任医师，南京医科大学附属南京医院骨科，江苏省南京市 210006 作者简介: 周建新，男，1982年生，江苏省吴江市人，汉族，南京医科大学在读硕士，主治医师，主要从事骨科创伤、关节研究。引用本文: 周建新,高峰,桂鉴超等. 组织工程化软骨修复界面整合的体外模型J. 中国组织工程研究, 2014, 18(27): 4324-4329. Zhou J