BMP-2 Gene Modified Canine bMSCs Promote Ectopic Bone Formation Mediated by a Nonviral PEI Derivative

被引：0
作者：
Kaige Lü
Deliang Zeng
Yilin Zhang
Lunguo Xia
Ling Xu
David L. Kaplan
Xinquan Jiang
Fuqiang Zhang
机构：
[1] Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University,Department of Prosthodontics
[2] School of Medicine,Oral Bioengineering Lab, Shanghai Research Institute of Stomatology
[3] Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University,Department of Biomedical Engineering
[4] School of Medicine,undefined
[5] Shanghai Key Laboratory of Stomatology,undefined
[6] Tufts University,undefined
来源：
Annals of Biomedical Engineering | 2011年 / 39卷
关键词：
Nonviral gene therapy; Polyethylenimine; Bone marrow stromal cells; Bone morphogenetic protein-2; Tissue engineering;
D O I：
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中图分类号：
学科分类号：
摘要：
The study was to explore the effects of BMP-2 gene modified canine bone marrow stromal cells (bMSCs) mediated by a nonviral PEI derivative (GenEscort™ II) in promoting bone formation in vitro and in vivo. Canine bMSCs were cultured and transfected with plasmids containing bone morphogenetic protein-2 gene (pBMP-2) or enhanced green fluorescent protein gene (pEGFP). Gene transfection conditions were initially optimized by varying GenEscort™ II/plasmid ratios. Osteogenic differentiation of gene modified bMSCs was investigated via alkaline phosphatase (ALP) activity analysis and real-time quantitative PCR (RT-qPCR) analysis in vitro. The bone formation ability of pBMP-2 transfected bMSCs combined with apatite-coated silk scaffolds (mSS) was explored and compared with pEGFP transfected bMSCs/mSS or untreated bMSCs/mSS at 8, 12 weeks after operation. Results showed that gene transfection efficiency reached up to 36.67 ± 4.12% as demonstrated by EGFP expression. ALP staining and activity assay were stronger with pBMP-2 gene transfection, and the mRNA expression of BMP-2, bone sialoprotein (BSP), Runt-related transcription factor 2 (Runx-2), and osteopontin (OPN) up-regulated in bMSCs 3, 6, 9 days in pBMP-2 group. Besides, the tissue-engineered bone complex with pBMP-2 modified bMSCs achieved significantly increased de novo bone formation compared with control groups (p < 0.01). We conclude that pBMP-2 transfection mediated by GenEscort™ II could enhance the osteogenic differentiation of canine bMSCs and promote the ectopic new bone formation in nude mice. GenEscort™ II mediated pBMP-2 gene transfer appears to be a safe and effective nonviral method for gene enhanced bone tissue engineering.
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页码：1829 / 1839
页数：10
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