Journal
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
Volume 103, Issue 12, Pages 3834-3842Publisher
WILEY-BLACKWELL
DOI: 10.1002/jbm.a.35527
Keywords
calcium phosphate; nanoparticles; hBMP-7 plasmid; transfection; osteogenesis; bone morphogenetic proteins
Funding
- Greek General Secretariat for Research and Technology Grant Thales-MIS [380278]
- Greek State Scholarships Foundation (IKY)
- Deutscher Akademischer Austauschdienst (DAAD)
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Functionalized calcium phosphate nanoparticles with osteogenic activity were prepared. Polyethyleneimine-stabilized calcium phosphate nanoparticles were coated with a shell of silica and covalently functionalized by silanization with thiol groups. Between the calcium phosphate surface and the outer silica shell, plasmid DNA which encoded either for bone morphogenetic protein 7 (BMP-7) or for enhanced green fluorescent protein was incorporated as cargo. The plasmid DNA-loaded calcium phosphate nanoparticles were used for the transfection of the pre-osteoblastic MC3T3-E1 cells. The cationic nanoparticles showed high transfection efficiency together with a low cytotoxicity. Their potential to induce an osteogenic response by transfection was demonstrated by measuring the alkaline phosphatase (ALP) activity and calcium deposition with alizarin red staining. The expression of the osteogenic markers Alp, Runx2, ColIa1 and Bsp was investigated by means of real-time quantitative polymerase chain reaction. It was shown that phBMP-7-loaded nanoparticles can provide a means of transient transfection and localized production of BMP-7 in MC3T3-E1 cells, with a subsequent increase of two osteogenic markers, specifically ALP activity and calcium accumulation in the extracellular matrix. Future strategies to stimulate bone regeneration focus into enhancing transfection efficiency and achieving higher levels of BMP-7 produced by the transfected cells. (c) 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 3834-3842, 2015.
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