中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (8): 1371-1376.doi: 10.3969/j.issn.1673-8225.2012.08.010

• 细胞外基质材料 extracellular matrix materials • 上一篇    下一篇

兔骨髓间充质干细胞与异体脱细胞耳软骨支架的体外复合*★

吴军成，吕仁荣，霍 然   

  1. 山东大学附属省立医院烧伤整形美容外科，山东省济南市  250021
  • 收稿日期:2011-11-03 修回日期:2011-12-19 出版日期:2012-02-19 发布日期:2012-02-19
  • 通讯作者: 霍然，博士，主任医师，博士生导师，山东大学附属省立医院烧伤整形美容外科，山东省济南市 250021
  • 作者简介:吴军成★，男，1979年生，山东省新泰市人，汉族，山东大学在读硕士，主治医师，主要从事软骨组织工程方面的研究。
  • 基金资助:

    山东省中青年科学家奖励基金计划项目(2004BS02009)，自体骨髓间充质干细胞和异体脱细胞软骨基质复合移植的研究。

In vitro composite of rabbit bone marrow mesenchymal stem cells and xenogenous acellular ear cartilage framework*★

Wu Jun-cheng, Lü Ren-rong, Huo Ran   

  1. Department of Burns and Plastic Surgery, Provincial Hospital Affiliated to Shandong University, Jinan  250021, Shandong Province, China
  • Received:2011-11-03 Revised:2011-12-19 Online:2012-02-19 Published:2012-02-19
  • Contact: Huo Ran, Doctor, Chief physician, Doctoral supervisor, Department of Burns and Plastic Surgery, Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China huoran@ mdemail.com.cn
  • About author:Wu Jun-cheng★, Studying for master’s degree, Attending physician, Department of Burns and Plastic Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China sdjnwjc@163.com
  • Supported by:

     the Reward Fund Project for Young and Middle-aged Scientists in Shandong Province, No. 2004BS02009*

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摘要：

背景：耳软骨作为脱细胞基质可选择的支架，进行脱细胞处理可去除了软骨细胞的抗原性，从而与种子细胞具有良好的相容性。
目的：体外提取、培养兔骨髓基质干细胞，并与异体脱细胞耳软骨支架复合，观察其生物相容性。
方法：提取兔骨髓间充质干细胞行体外培养，诱导为软骨细胞，以胰蛋白酶-曲拉通联合法获得脱细胞软骨支架，将两者于体外复合，10 d后复合支架固定行组织学染色及扫描电镜观察。
结果与结论：兔骨髓间充质干细胞体外诱导14 d可形成软骨细胞，Ⅱ型胶原免疫细胞化学示胞浆呈棕黄色；兔耳软骨脱细胞基质呈乳白色，组织学染色及扫描电镜观察示经脱细胞后支架孔隙均匀，结构完整，仍保存大量酸性黏多糖及胶原成分。其孔径长度(33.70±4.33) μm，孔隙率(65.23±7.35)%。
复合支架组织学染色及扫描电镜示两者黏附良好，并伴有多量基质分泌。说明兔骨髓间充质干细胞诱导为软骨细胞与异体脱细胞耳软骨有良好的生物相容性。

关键词: 脱细胞耳软骨, 骨髓间充质干细胞, 软骨细胞, 复合支架, 生物相容性

Abstract:

BACKGROUND: Ear cartilage, an optional framework for acellular matrix, loses its antigenicity to chondrocytes after acellular treatment; therefore it has a good compatibility with seed cells.
OBJECTIVE: To extract and culture rabbit bone marrow mesenchymal stem cells; to composite the stem cells with xenogenous acellular ear cartilage framework; and to explore the biocompatibility between them both.
METHODS: Rabbit bone marrow mesenchymal stem cells were isolated, cultured in vitro and induced into cartilage cells. The acellular cartilage framework was obtained by trypsin-Triton combined method; then the two were composited in vitro. The composited framework was fixed for histological staining and scanning electron microscopic observation on 10 day after composite.
RESULTS AND CONCLUSION: Rabbit bone marrow mesenchymal stem cells were induced into cartilage cells on 14 day after  in vitro induction. Type Ⅱ collagen immunohistochemistry showed the cytoplasm was brownish yellow; rabbit ear acellular cartilage matrix was ivory white. Histological staining and scanning electron microscopy indicated that the pores were evenly distributed on the framework after acellular treatment; the framework has complete structure with massive acid mucopolysaccharide and collagen components. The pore size was (33.70±4.33) μm; the porosity was (65.23±7.35)%. Histological staining and scanning electron microscopy of the composite framework showed good adhesion between the two, accompanied by large amounts of matrix secretion. These findings indicate that the chondrocytes induced from rabbit bone marrow mesenchymal stem cells have a good biocompatibility with allogenic acellular ear cartilage.

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