• Title/Summary/Keyword: non-viral gene delivery

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Preparation and In Vitro Release of DNA-Loaded Poly(D,L-lactic-co-glycolic acid) Microspheres (DNA가 봉입된 Poly(D,L-lactic-co-glycolic acid) 미립구의 제조 및 시험관내 방출)

  • Son, Hye-Jung;Kim, Jin-Seok
    • Polymer(Korea)
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    • v.29 no.1
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    • pp.69-73
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    • 2005
  • To overcome the main disadvantages of non-viral gene delivery systems such as repeated administration due to the low transfection efficiency, poly(D,L-lactide-co-glycolide) was applied to encapsulate pDNA in its microsphere formulation. Free pDNA or various ratios (w/w) of chitosan/pDNA complexes was used for encapsulation, with the resulting encapsulation efficiency of 44%, 5%, and 8% for free pDNA, 0.7:1 and 1:1 ratios, respectively. Scanning electron micrographs of poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres encapsulating pDNA or chitosan-condensed pDNA revealed a smooth spherical shape immediately after microsphere preparation and a collapsed porous shape in 41 days due to the degradation of PLGA. In vitro release profile showed that the 0.7:1 (w/w) ratio formulation exerted 47% release in 26 days, whereas free pDNA or 1:1 (w/w) ratio formulation did only 15% or 32%, respectively.

Osteogenic effects of polyethyleneimine-condensed BMP-2 genes in vitro and in vivo (Polyethyleneimine-응축 BMP-2 발현 유전자를 이용한 골형성 효과)

  • Cheong, Hee-Sun;Kim, Kyoung-Hwa;Park, Yoon-Jeong;Kim, Tae-Il;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul;Lee, Dong-Soo;Lee, Seung-Jin;Chung, Chong-Pyoung;Han, Soo-Boo;Seol, Yang-Jo
    • Journal of Periodontal and Implant Science
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    • v.37 no.4
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    • pp.859-869
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    • 2007
  • 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.