中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (38): 7031-7036.doi: 10.3969/j.issn.1673-8225.2010.38.001

• 组织工程骨及软骨材料 tissue-engineered bone and cartilage materials •    下一篇

体外构建腺病毒介导骨形态发生蛋白2基因转染骨髓间充质干细胞复合纳米羟基磷灰石的植骨材料

黄俊锋1,2，王大平2，何春耒1，杨欣建2，刘黎军2，陈宏贤2，彭亮权2   

  1. 1广州医学院，广东省广州市  510182；2深圳市第二人民医院创伤骨科，广东省深圳市  518035
  • 出版日期:2010-09-17 发布日期:2010-09-17
  • 通讯作者: 王大平，教授，主任医师，博士，深圳市第二人民医院创伤骨科，广东省深圳市 518035 dapingwang@medmail.com.cn
  • 作者简介:黄俊锋★，男，1977年生，广东省潮州市人，汉族，2002年中山医科大学毕业，硕士，主治医师，主要从事创伤骨科、手外科临床工作。hjf097@163.com
  • 基金资助:

    深圳市卫生局科技计划项目资助(200630)。

Adenovirus mediated bone morphogenetic protein-2 gene transfection of bone marrow mesenchymal stem cells combined with nano-hydroxyapatite to construct bone graft material in vitro

Huang Jun-feng 1,2, Wang Da-ping2, He Chun-lei1, Yang Xin-jian2, Liu Li-jun2, Chen Hong-xian2, Peng Liang-quan2   

  1. 1 Guangzhou Medical College, Guangzhou   510182, Guangdong Province, China; 2 Department of Orthopaedics, Shenzhen Second People’s Hospital, Shenzhen   518035, Guangdong Province, China
  • Online:2010-09-17 Published:2010-09-17
  • Contact: Wang Da-ping, Professor, Chief physician, Doctor, Department of Orthopaedics, Shenzhen Second People’s Hospital, Shenzhen 518035, Guangdong Province, China dapingwang@medmail.com.cn
  • About author:Huang Jun-feng★, Master, Attending physician, Guangzhou Medical College, Guangzhou 510182, Guangdong Province, China; Department of Orthopaedics, Shenzhen Second People’s Hospital, Shenzhen 518035, Guangdong Province, China hjf097@163.com
  • Supported by:

    Science and Technology Projects by Health Bureau of Shenzhen, No. 200630*

PDF

457

被引次数

3

摘要：

背景：随着组织工程和基因工程技术迅速发展，基因增强的组织工程骨为临床治疗骨缺损带来美好的前景。
目的：观察经腺病毒介导的人骨形态发生蛋白2表达载体(Ad-BMP-2)转染后的兔骨髓间充质干细胞在纳米羟基磷灰石植骨材料上的黏附、增殖及骨形态发生蛋白2的表达情况。
方法：用Ad-BMP-2转染兔骨髓间充质干细胞，免疫组化、蛋白印迹法检测转染后细胞内骨形态发生蛋白2的表达情况。将转染48 h的细胞均匀接种到纳米羟基磷灰石植骨材料上，扫描电镜观察细胞黏附状况，并采用MTT比色法测定骨髓间充质干细胞的增殖情况；消化收集黏附植骨材料上第3，5，7天的细胞，蛋白印迹检测黏附材料上细胞骨形态发生蛋白2的表达。
结果与结论：转染后，骨髓间充质干细胞内骨形态发生蛋白2有高表达；扫描电镜见转染细胞在纳米羟基磷灰石材料孔隙周围及孔隙内黏附生长良好并大量增殖，MTT分析结果显示，纳米羟基磷灰石对骨髓间充质干细胞的体外增殖无抑制作用，复合后骨形态发生蛋白2有较高表达。结果表明经Ad-BMP-2转染后的骨髓间充质干细胞与纳米羟基磷灰石植骨材料生物相容性好，细胞在材料上能稳定长效地分泌骨形态发生蛋白2。

关键词: 腺病毒载体, 骨形态蛋白, 纳米羟基磷灰石, 骨髓间充质干细胞, 组织工程骨

Abstract:

BACKGROUND: With advanced tissue engineering and gene engineering technologies, gene enhanced tissue engineered bone provides prosperous application for the repair of bone deficits. 
OBJECTIVE: To study the adhesion, proliferation and expression of bone marrow mesenchymal stem cells (BMSCs) on nano-hydroxyapatite (Nano-HA) bone graft material after transfection of adenovirus-mediated human bone morphogenetic protein-2 expression vector (Ad-BMP-2).  
METHODS: BMSCs were transfected using Ad-BMP-2. Immunohistochemistry and Western blot were used to detect BMP-2 expression in transfected cells. After transfected 48 hours, cells were inoculated on Nano-HA bone graft material evenly, scanning electron microscopy was used to observe the adhesion status of cells on material, and MTT test was used to assay the proliferation of transfected BMSCs. Cells adhered on material were gathered on days 3, 5, 7, Western blot was used to detect the expression of BMP-2 of the adhered cells. 
RESULTS AND CONCLUSION: After transfection, BMP-2 protein was highly expressed in BMSCs; Using scanning electron microscopy, the transfected BMSCs were seen to grow well around or in pores of the Nano-HA bone graft material, and proliferated. MTT test assay showed that, the Nano-HA bone graft material couldn’t inhibit in vitro proliferation of BMSCs. The transfection cells adhered on composite had a higher BMP-2 protein expression. Results demonstrated that Ad-BMP-2-transfected BMSCs are well biocompatible to Nano-HA bone graft material, the transfected cells in material can secrete BMP-2 stably for a long time.

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