title: BMP-2 gene modified canine bMSCs promote ectopic bone formation mediated by a nonviral PEI derivative.
abstract: 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.
authors.affiliation: Department of Prosthodontics, Ninth People's Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai, 200011, People's Republic of China.
authors.forename: Kaige
authors.initials: K
authors.lastname: Lü
authors.forename: Deliang
authors.initials: D
authors.lastname: Zeng
authors.forename: Yilin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Lunguo
authors.initials: L
authors.lastname: Xia
authors.forename: Ling
authors.initials: L
authors.lastname: Xu
authors.forename: David L
authors.initials: DL
authors.lastname: Kaplan
authors.forename: Xinquan
authors.initials: X
authors.lastname: Jiang
authors.forename: Fuqiang
authors.initials: F
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D016026:Bone Marrow Transplantation; D055396:Bone Morphogenetic Protein 2 / Q000096:biosynthesis* / Q000235:genetics; D002454:Cell Differentiation; D004285:Dogs; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D010012:Osteogenesis; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection; D014183:Transplantation, Heterologous
chemical_list: D000943:Antigens, Differentiation; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2