title: Ectopic study of calcium phosphate cement seeded with pBMP-2 modified canine bMSCs mediated by a non-viral PEI derivative.
abstract: We have evaluated the ectopic new bone formation effects of CPC (calcium phosphate cement) seeded with pBMP-2 (plasmids containing bone morphogenetic protein-2 gene) transfected canine bMSCs (bone marrow stromal cells) mediated by a non-viral PEI (polyethylenimine) derivative (GenEscort™ II) in nude mice. Canine bMSCs were transfected with pBMP-2 or pEGFP (plasmids containing enhanced green fluorescent protein gene) mediated by GenEscort™ II in vitro, and the osteoblastic differentiation was explored by ALP (alkaline phosphatase) staining, ARS (alizarin red S) staining and RT-qPCR (real-time quantitative PCR) analysis. Ectopic bone formation effects of CPC/pBMP-2 transfected bMSCs were evaluated and compared with CPC/pEGFP transfected bMSCs or CPC/untransfected bMSCs through histological, histomorphological and immunohistochemical analysis 8 and 12 weeks post-operation in nude mice. Transfection efficiency was up ∼35% as demonstrated by EGFP (enhanced green fluorescent protein) expression. ALP and ARS staining were stronger with pBMP-2 gene transfection, and mRNA expression of BMP-2 (bone morphogenetic protein-2), Col 1 (collagen 1) and OCN (osteocalcin) in pBMP-2 group was significantly up-regulated at 6 and 9 days. Significantly higher NBV (new bone volume) was achieved in pBMP-2 group than in the control groups at 8 and 12 weeks (P<0.05). In addition, immunohistochemical analysis indicated higher OCN expression in pBMP-2 group (P<0.01). We conclude that CPC seeded with pBMP-2 transfected bMSCs mediated by GenEscort™ II could enhance ectopic new bone formation in nude mice, suggesting that GenEscort™ II mediated pBMP-2 gene transfer is an effective non-viral method and CPC is a suitable scaffold for gene enhanced bone tissue engineering.
authors: Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Peoples Republic of China.
authors: Kaige
authors: K
authors: Lü
authors: Deliang
authors: D
authors: Zeng
authors: Wenjie
authors: W
authors: Zhang
authors: Lunguo
authors: L
authors: Xia
authors: Ling
authors: L
authors: Xu
authors: Xinquan
authors: X
authors: Jiang
authors: Fuqiang
authors: F
authors: Zhang
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D001842:Bone and Bones / Q000473:pathology; D002130:Calcium Phosphates / Q000378:metabolism*; D049109:Cell Proliferation; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D005786:Gene Expression Regulation; D008297:Male; D051379:Mice; D008819:Mice, Nude; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011094:Polyethyleneimine / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D023822:Tissue Engineering; D014162:Transfection
chemical_list: D055396:Bone Morphogenetic Protein 2; D002130:Calcium Phosphates; D024042:Collagen Type I; D015675:Osteocalcin; D011094:Polyethyleneimine; C020243:calcium phosphate