摘要
目的观察腺病毒介导的人骨形态发生蛋白绿色荧光蛋白基因（Ad-GFP-hBMP-2）转染对骨髓间质干细胞（bMSCs）成骨能力的影响。方法取日本大耳白兔4只自双侧股骨远端抽取骨髓培养bMSCs。以Ad—GFP—hBMP-2基因（实验组）及Ad—GFP（对照组）基因转染bMSCs后，用ALP检测试剂盒检测两组细胞的ALP活性；原位杂交检测两组细胞Ⅰ型胶原的表达；Westernblot检测细胞中BMP-2的表达。将转染后24h的bMSCs接种到裸鼠体内，术后第4、8、12周观察成骨情况。结果转Ad—GFP—hBMP-2基因组和Ad—GFP组各时间段ALP分泌量差异分别有统计学意义（P〈0．01）；实验组Ⅰ型胶原原位杂交实验组为阳性。实验组成骨阳性率为90％，对照组为40％。结论bMSCs经Ad—GFP—hBMP-2基因转染后能高效表达BMP-2并诱导成骨。腺病毒介导人BMP-2转基因可以提高bMSCs的成骨能力。
Objective To evaluate the osteogenic potential of bone mesenchymal stem cells （bM- SCs） transfected with gene Ad-GFP-hBMP-2. Methods MSCs obtained from dual femoral of Japan big ear rabbits were infected with Ad-GFP-hBMP-2 as experimental group and Ad-GFP as control group. ALP kit was used to measure ALP activity, and hybridization in situ kit was used to observe the mRNA synthesis of collagen type Ⅰ. After transfection,5.0 ×10^6 cells were transplanted into the muscle of nude mice. Then animals were observed by X-ray, and specimens that were removed 4,8, and 12 weeks postoperatively underwent histological examination. Results After treated with the gene Ad-GFP-hBMP-2, the bMSCs synthesized BMP-2 successfully and gained the characters similar to those of osteoblasts, and bone was formed in nude mice. Conclusion Both GFP and BMP-2-pmducing bMSCs created by means of adenoviral gene transfer produce BMP-2 to induce new bone formation, and express GFP to track the cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第12期1690-1692,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30672125）
关键词
骨
组织工程
基因转染
骨髓间充质干细胞
骨形态发生蛋白-2
Bone
Tissue engineering
Gene transfection
Bone marrow mesenchymal stem ceils
Bone morphogenetic protein-2
