J Korean Acad Periodontol.  2007 Dec;37(4):859-869. 10.5051/jkape.2007.37.4.859.

Osteogenic effects of polyethyleneimine-condensed BMP-2 genes in vitro and in vivo

Affiliations
  • 1Department of Periodontology School of Dentistry Seoul National University, Korea. yjseol@snu.ac.kr
  • 2BK21 Craniomaxillofacial Life Science School of Dentistry Seoul National University, Korea.
  • 3Craniomaxillofacial Reconstructive Science School of Dentistry Seoul National University, Korea.
  • 4Department of Nuclear Medicine, College of Medicine Seoul National University, Korea.
  • 5College of Pharmacy Ewha Womans University, Korea.

Abstract

Naked DNA and standard vectors have been previously used for gene delivery. Among these, PEI can efficiently condense DNA and has high intrinsic endosomal activities. The aim of this study is to investigate whether the cationic polycation PEI could increase the transfection efficiency of BMP expressing DNA using a vector-loaded collagen sponge model. BMP-2/pcDNA3.1 plasmid was constructed by subcloning human BMP-2 cDNA into the pcDNA3.1 plasmid vector. PEI/DNA complexes were prepared by mixing PEI and BMP-2/pcDNA3.1 and the constructed complexes were loaded into the collagen sponges. In vitro studies, BMSCs were transfected with the PEI/BMP-2/pcDNA3.1 complexes from collgen sponge. The level of secreted BMP-2 and alkaline phosphatase activities of transfected BMSCs were significantly higher in PEI/BMP-2/pcDNA3.1 group than in BMP-2/pcDNA3.1 group (p<0.05). Transfected BMSCs were cultured and mineralization was observed only in cells treated with PEI/BMP-2/pcDNA3.1 complexes. In vivo studies, PEI/BMP-2/pcDNA3.1/collagen, BMP-2/pcDNA3.1/collagen and blank collagen were grafted in skeletal muscle of nude mice. Ectopic bone formation was shown in PEI/BMP-2/pcDNA3.1/collagen grafted mouse 4 weeks postimplantation, while not in BMP-2/pcDNA3.1 grafted tissue. This study suggests that PEI-condensed DNA encoding for BMP-2 is capable of inducing bone formation in ectopic site and might increase the transfection rate of BMP-2/pcDNA3.1. As a non-viral vector, PEI offers the potential in gene therapy for bone engineering.

Keyword

Gene therapy; BMP; PEI; Vector; DNA; Bone formation

MeSH Terms

Alkaline Phosphatase
Animals
Collagen
DNA
DNA, Complementary
Genetic Therapy
Humans
Mice
Mice, Nude
Muscle, Skeletal
Osteogenesis
Plasmids
Porifera
Transfection
Transplants
Alkaline Phosphatase
Collagen
DNA
DNA, Complementary

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