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Calcium phosphate nanoparticles carrying BMP‐7 plasmid DNA induce an osteogenic response in MC3T3‐E1 pre‐osteoblasts
Author(s) -
Hadjicharalambous Chrystalleni,
Kozlova Diana,
Sokolova Viktoriya,
Epple Matthias,
Chatzinikolaidou Maria
Publication year - 2015
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.35527
Subject(s) - transfection , alkaline phosphatase , bone morphogenetic protein 2 , calcium , materials science , biophysics , osteoblast , microbiology and biotechnology , biochemistry , chemistry , in vitro , biology , enzyme , metallurgy , gene
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. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 3834–3842, 2015.