Osteogenic differentiation of rat bone marrow mesenchymal stem cells induced by bone morphogenetic protein gene 2 vector
GAO Jun-huai, QIU Su-jun, YIN Jie, SHI Ben-chao, MIN Shao-xiong
Chinese Journal of Clinical Anatomy ›› 2018, Vol. 36 ›› Issue (2) : 187-191.
Osteogenic differentiation of rat bone marrow mesenchymal stem cells induced by bone morphogenetic protein gene 2 vector
Objective To study the effect of acetylcysteine/phosphorylcholine-modified chitosan (NAC-PCCs) encapsulation of bone morphogenetic protein gene 2 nanoparticle materials in promoting the osteogenic differentiation of SD rat bone marrow mesenchymal stem cells (BMSC) in vitro. Methods The pBMP2/NAC-PCCs nanoparticles were prepared, and its characterization, in vitro release and the ability to protect bone morphogenetic protein gene 2 against DNaseⅠwere examined. BMSC cells, nanoparticles in the process of cultivation, and transfection efficiency were determined by fluorescence microscopy and flow cytometry. The secretion of BMP2 protein levels was determined by ELISA experiment and the expression of RUNX2 and OC gene was determined by RT-PCR. The calcium deposition analysis was performed using alizarin red staining. Results Gel electrophoresis showed that nanoparticles could effectively protect DNA from being digested by DNaseⅠ. Fluorescent microscopy and flow cytometry showed that the gene delivery nanoparticles had better transfection efficiency. The results of ROS activity test showed that the material had the ability to scavenge ROS, and the ELISA experiment showed that the expression level of BMP2 in the group of pBMP2/NAC-PCCs was the highest within 14 days. QRT-PCR, AKP activity detection and alizarin red staining showed that the osteogenic differentiation of pBMP2/NAC-PCC was the best. Conclusion pBMP2/NAC-PCCs nanoparticles can promote the osteogenic differentiation of BMSC.
Repair of bone injury / Non-viral vector / Osteogenic differentiation / Bone morphogenetic protein gene 2
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