摘要
目的:体外观察碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)联合Sonic Hedgehog(Shh)的时序性基因传递对骨髓来源的间充质干细胞(bone marrow derived mesenchymal stem cells,BMSCs)中成骨相关因子的影响。方法:将前期实验中构建的重组病毒rAAV2-tet-off-bFGF及rAAV2-Shh感染原代培养的BMSCs,其中前者可被四环素调控系统调控bFGF的表达。real-time RT-PCR及Western blot检测各成骨相关因子的表达变化。结果:双病毒感染BMSCs后可被四环素衍生物强力霉素(doxycycline,Dox)调控。细胞内成骨相关因子的mRNA及蛋白水平较对照组都有不同程度的升高,变化具有统计学差异。结论:短期bFGF的表达联合Shh的基因传递能极大程度提高BMSCs的增殖能力,增加胞内成骨相关因子的表达水平,促进BMSCs成骨分化。
Objective： The aim of this study was to delivery basic fibroblast growth factor （bFGF） and sonic hedgehog （Shh） to bone marrow derived mesenchymal stem cells （BMSCs）, and then to detect the effect for osteogenic marker genes. Method： bFGF and Shh were constructed to the recombinant adeno-associated virus, respectively （rAAV2-tet-off-bFGF and rAAV2-Shh）. The previous viral vector allowed for regulation of the bFGF expression by the addition of doxycycline, a tetracycline analogue. Several osteogenic markers were detected by quantitative real-time reverse transcriptase polymerase chain reaction and western blot. Result： BMSCs were constructed successfully by two recombinant virus that were regulated by Dox. The expression of osteogenic marker mRNA and protein had an increased tendency after two genes transduction （P 〈 0.05）. Conclusion： Sequential delivery of angiogenic and osteogenic factors likely has a synergistic effect, which could not only enhance the cell proliferation, but also improve the expression of osteogenic marker of BMSCs.
出处
《临床口腔医学杂志》
2015年第9期521-526,共6页
Journal of Clinical Stomatology
基金
国家自然科学基金(81400490)
湖北省自然科学基金(2014CFB388)
