BMP-2 Gene Modified Canine bMSCs Promote Ectopic Bone Formation Mediated by a Nonviral PEI Derivative
The study was to explore the effects of BMP-2 gene modified canine bone marrow stromal cells (bMSCs) mediated by a nonviral PEI derivative (GenEscortTM 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 phos-phatase (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 tissueengineered 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 Gen Escort? 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.
Nonviral gene therapy Polyethylenimine Bone marrow stromal cells Bone morphogenetic protein-2 Tissue engineering.
Kaige Lu Deliang Zeng Yilin Zhang Lunguo Xia Ling Xu David L. Kaplan Xinquan Jiang Fuqiang Zhang
Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, S Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, S Oral Bioengineering Lab, Shanghai Research Institute of Stomatology, Ninth Peoples Hospital Affilia Department of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA 02155, USA
国际会议
上海
英文
181-191
2011-12-28（万方平台首次上网日期，不代表论文的发表时间）