Ectopic study of calcium phosphate cement seeded with pBMP-2 modified canine bMSCs mediated by a non-viral PEI derivative

Authors

  • Kaige Lü,

    1. Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
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  • Deliang Zeng,

    1. Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
    2. Oral Bioengineering Lab, Shanghai Research Institute of Stomatology, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Key Laboratory of Stomatology, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
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  • Wenjie Zhang,

    1. Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
    2. Oral Bioengineering Lab, Shanghai Research Institute of Stomatology, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Key Laboratory of Stomatology, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
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  • Lunguo Xia,

    1. Oral Bioengineering Lab, Shanghai Research Institute of Stomatology, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Key Laboratory of Stomatology, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
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  • Ling Xu,

    1. Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
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  • Xinquan Jiang,

    Corresponding author
    1. Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
    2. Oral Bioengineering Lab, Shanghai Research Institute of Stomatology, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Key Laboratory of Stomatology, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
      Correspondence may be addressed to either of these authors (email xinquanj@yahoo.cn or fqzhang@vip.163.com)
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  • Fuqiang Zhang

    Corresponding author
    1. Department of Prosthodontics, Ninth Peoples Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 639 Zhizaoju Road, Shanghai 200011, Peoples Republic of China
      Correspondence may be addressed to either of these authors (email xinquanj@yahoo.cn or fqzhang@vip.163.com)
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Correspondence may be addressed to either of these authors (email xinquanj@yahoo.cn or fqzhang@vip.163.com)

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.

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