| 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 |
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Received:October 29, 2003
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| 期刊信息：《中国骨伤》2004年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) |
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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 |
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| Construction of tissue engineering bone with hBMP-2 gene transfected bone marrow stromal cells and bovine cancellous bone scaffold in vitro |
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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. |
| KEYWORDS:Bone morphogenetic protein Bone marrow stromal cells Gene transfection Tissue engineering bone |
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| 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 |
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