title: Calcium phosphate nanoparticles carrying BMP-7 plasmid DNA induce an osteogenic response in MC3T3-E1 pre-osteoblasts.
abstract: 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.
authors.affiliation: Department of Materials Science and Technology, University of Crete, P.O. Box 2208, Heraklio, 71001, Greece.
authors.forename: Chrystalleni
authors.initials: C
authors.lastname: Hadjicharalambous
authors.affiliation: Institute of Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitaetsstr. 5-7, Essen, 45117, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Kozlova
authors.affiliation: Institute of Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitaetsstr. 5-7, Essen, 45117, Germany.
authors.forename: Viktoriya
authors.initials: V
authors.lastname: Sokolova
authors.affiliation: Institute of Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitaetsstr. 5-7, Essen, 45117, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Epple
authors.affiliation: Department of Materials Science and Technology, University of Crete, P.O. Box 2208, Heraklio, 71001, Greece.|Institute of Electronic Structure and Laser (IESL), Foundation for Research and Technology Hellas (FORTH), N. Plastira 100, Heraklio, 70013, Greece.
authors.forename: Maria
authors.initials: M
authors.lastname: Chatzinikolaidou
mesh_terms: D000818:Animals; D055419:Bone Morphogenetic Protein 7 / Q000235:genetics*; D002130:Calcium Phosphates / Q000737:chemistry*; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D006801:Humans; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D010012:Osteogenesis; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection
chemical_list: C525819:BMP7 protein, human; D055419:Bone Morphogenetic Protein 7; D002130:Calcium Phosphates; D004247:DNA; C020243:calcium phosphate
keywords: bone morphogenetic proteins; calcium phosphate; hBMP-7 plasmid; nanoparticles; osteogenesis; transfection