NAC-PCCs载骨形态发生蛋白2基因纳米微球促大鼠骨髓间充质干细胞成骨分化

doi:10.13418/j.issn.1001-165x.2018.02.013

Chinese Journal Of Clinical Anatomy ›› 2018, Vol. 36 ›› Issue (2): 187-191.doi: 10.13418/j.issn.1001-165x.2018.02.013

Previous Articles Next Articles

Osteogenic differentiation of rat bone marrow mesenchymal stem cells induced by bone morphogenetic protein gene 2 vector

GAO Jun-huai¹, QIU Su-jun¹, YIN Jie², SHI Ben-chao¹, MIN Shao-xiong¹

1. Department of Spinal Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China; 2. Ningbo No.6 Hospital, Ningbo 315040, China

Received:2018-02-28 Online:2018-03-25 Published:2018-05-04

Abstract

Abstract:

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.

Key words: Repair of bone injury, 　Non-viral vector, 　Osteogenic differentiation, 　Bone morphogenetic protein gene 2

GAO Jun-huai, QIU Su-jun, YIN Jie, SHI Ben-chao, MIN Shao-xiong. Osteogenic differentiation of rat bone marrow mesenchymal stem cells induced by bone morphogenetic protein gene 2 vector[J]. Chinese Journal Of Clinical Anatomy, 2018, 36(2): 187-191.

References

[1] Jin H, Zhang K, Qiao C, et al. Efficiently engineered cell sheet using a complex of polyethylenimine-alginate nanocomposites plus bone morphogenetic protein 2 gene to promote new bone formation[J]. Int J Nanomedicine, 2014. 9: 2179-2190.

[2] Calori GM, Donati D, Di Bella C, et al. Bone morphogenetic proteins and tissue engineering: future directions[J]. Injury, 2009, 3: S67-76.
[3] Zigdongiladi H, Rudich U, Geller GM, et al. Recent advances in bone regeneration using adult stem cells[J]. World J Stem Cells, 2015, 7(3): 630.
[4] Gnecchi M, Danieli P, Malpasso G, et al. Paracrine Mechanisms of Mesenchymal Stem Cells in Tissue Repair[J]. Methods in Molecular Biology, 2016, 1416: 123-146.
[5] Wang W, Sun L, Zhang P, et al. An anti-inflammatory cell-free collagen/resveratrol scaffold for repairing osteochondral defects in rabbits[J]. Acta Biomater, 2014, 10(12): 4983-4995.
[6] Wang FW, Wang Z, Zhang YM, et al. Protective effect of melatonin on bone marrow mesenchymal stem cells against hydrogen peroxide-induced apoptosis in vitro[J]. Journal of Cellular Biochemistry, 2013, 114(10): 2346-2355.
[7] Malakooty Poor E, Baghaban Eslaminejad M, Gheibi N, et al. Chitosan/DNA nanoparticles characteristics determine the transfection efficacy of gene delivery to human mesenchymal stem cells[J]. Artif Cells Nanomed Biotechnol，2014，42(6):376-384.
[8] Li Y, Xue F,Xu SZ, et al. Lycopene protects bone marrow mesenchymal stem cells against ischemia-induced apoptosis in vitro[J]. Eur Rev Med Pharmacol Sci, 2014, 18(11): 1625-1631.
[9] Min SL, Kim NW, Lee K, et al. Enhanced transfection by antioxidative polymeric gene carrier that reduces polyplex-mediated cellular oxidative stress[J]. Pharmaceutical Research, 2013, 30(6): 1642-1651.
[10]Pramanik S, Pingguanmurphy B, Cho J, et al. Design and development of potential tissue engineering scaffolds from structurally different longitudinal parts of a bovine-femur[J]. Sci Rep, 2014, 4:5843.
[11]Marycz K, Mieszek A, Grzesiak J, et al. The osteogenic properties of multipotent mesenchymal stromal cells in cultures on TiO2 sol-gel-derived biomaterial[J]. Biomed Res Int, 2015, 2015:651097.
[12]Evans CH. Gene therapy for bone healing[J]. Expert Rev Mol Med, 2010. 6(12)：18.

Osteogenic differentiation of rat bone marrow mesenchymal stem cells induced by bone morphogenetic protein gene 2 vector

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 0

Metrics

Comments

Recommended 0