重组PcDNA3.1-hBMP-2转染骨髓基质干细胞及复合异种骨支架体外构建组织工程骨
摘要点击次数: 2115   全文下载次数: 1383   投稿时间:2003-10-29    
作者Author单位AddressE-Mail
卜丽莎 BU Li-sha 吉林大学中日联谊医院中心研究室,吉林长春130021 The China-Japan Union Hospital of Jilin University bulisha@163.com 
李建军 LI Jian-jun 吉林大学第一医院骨科  
韩东 HAN Dong 吉林大学中日联谊医院手外科  
景元海 JING Yuan-hai 吉林大学中日联谊医院骨科  
王宏 WANG Hong 吉林大学第一医院骨科  
王玲 WANG Ling 吉林大学白求恩医学院分子生物学研究室  
徐莘香 XU Shen-xiang 吉林大学第一医院骨科  
期刊信息:《中国骨伤》2004年,第17卷,第8期,第449-451页
DOI:doi:10.3969/j.issn.1003-0034.yyyy.nn.zzz
基金项目:国家自然科学基金资助项目(39800151);吉林省科技厅基金资助项目(20010110)
中文摘要:

目的:构建人骨形态发生蛋白2(humanbonemorphogeneticprotein,hBMP-2)真核表达载体PcDNA31hBMP-2,转染兔骨髓基质干细胞(bonemarrowstromalcells,BMSCs),种植去抗原牛松质骨(bovinecancellousbone,BCB)支架体外构建组织工程骨。

方法:蛋白印迹法检测转染后细胞BMP-2的表达,碱性磷酸酶(ALPase)活性检测分析基因转染对细胞分化的影响。然后将转染后细胞接种到BCB支架上,扫描电镜观察细胞贴附、生长状况。

结果:转染后,BMSCs表达BMP-2,ALP活性明显增高。扫描电镜见转染细胞分布均匀,伸展良好。

结论:在脂质体介导下,BMP-2基因可导入细胞且稳定表达基因产物促进自身增殖分化,转染后细胞在支架材料上贴附生长良好,为进一步应用携带BMP-2基因的人工骨修复骨缺损奠定了实验基础。
【关键词】骨形态发生蛋白  骨髓基质干细胞  基因转染  组织工程骨
 
Construction of tissue engineering bone with hBMP-2 gene transfected bone marrow stromal cells and bovine cancellous bone scaffold in vitro
ABSTRACT  

Objective:To construct rabbit bone marrow stromal cells (BMSCs) transfecting by a recombinant plasmid carrying the human bone morphogenetic protein-2 gene (PcDNA3.1-hBMP-2) seeded into bovine cancellous bone (BCB) scaffolds to construct tissue engineering bone in vitro.

Methods:The expression of hBMP-2 in these cells after transfection was determined by Western blot analysis.The changes of cellular differentiation were observed by ALPase activity analysis.BMP-2 transduced cells were then seeded into BCB scaffolds.The attachment and growth of the cells on the scaffold were examined using scanning electron microscope (SEM).

Results:The expression of hBMP-2 was confirmed and ALPase activity obviously increased in the cells after transfection.SEM examination revealed extensive cellular attachment and growth on the BCB block.

Conclusion:With the help of lipofectamine,the transfection of PcDNA3.1-hBMP-2 to BMSCs is carried out successfully.The cells after transfection grow well on a BCB scaffold.Tissue engineering bone used to regional gene therapy is constructed successfully.
KEY WORDS  Bone morphogenetic protein  Bone marrow stromal cells  Gene transfection  Tissue engineering bone
 
引用本文,请按以下格式著录参考文献:
中文格式:卜丽莎,李建军,韩东,景元海,王宏,王玲,徐莘香.重组PcDNA3.1-hBMP-2转染骨髓基质干细胞及复合异种骨支架体外构建组织工程骨[J].中国骨伤,2004,17(8):449~451
英文格式:BU Li-sha,LI Jian-jun,HAN Dong,JING Yuan-hai,WANG Hong,WANG Ling,XU Shen-xiang.Construction of tissue engineering bone with hBMP-2 gene transfected bone marrow stromal cells and bovine cancellous bone scaffold in vitro[J].zhongguo gu shang / China J Orthop Trauma ,2004,17(8):449~451
阅读全文  下载  查看/发表评论  下载PDF阅读器
关闭




版权所有:《中国骨伤》杂志社京ICP备12048066号-2  版权声明
地址:北京市东直门内南小街甲16号,100700
电话:010-64089487 传真:010-64089792 Email:zggszz@sina.com

京公网安备 11010102004237号