title: Non-viral bone morphogenetic protein 2 transfection of rat dental pulp stem cells using calcium phosphate nanoparticles as carriers.
abstract: Calcium phosphate nanoparticles have shown potential as non-viral vectors for gene delivery. The aim of this study was to induce bone morphogenetic protein (Bmp)2 transfection in rat dental pulp stem cells using calcium phosphate nanoparticles as a gene vector and then to evaluate the efficiency and bioactivity of the transfection. We also intended to investigate the behavior of transfected cells when seeded on 3-dimensional titanium fiber mesh scaffolds. Nanoparticles of calcium phosphate encapsulating plasmid deoxyribonucleic acid (DNA) (plasmid enhanced green fluorescent protein-BMP2) were prepared. Then, STRO-1-selected rat dental pulp stem cells were transfected using these nanoparticles. Transfection and bioactivity of the secreted BMP2 were examined. Thereafter, the transfected cells were cultured on a fibrous titanium mesh. The cultures were investigated using scanning electron microscipy and evaluated for cell proliferation, alkaline phosphatase activity and calcium content. Finally, real-time polymerase chain reaction was performed for odontogenesis-related gene expression. The results showed that the size of the DNA-loaded particles was approximately 100 nm in diameter. Nanoparticles could protect the DNA encapsulated inside from external DNase and release the loaded DNA in a low-acid environment (pH 3.0). In vitro, nanoparticle transfection was shown to be effective and to accelerate or promote the odontogenic differentiation of rat dental pulp stem cells when cultured in the 3-dimensional scaffolds. Based on our results, plasmid DNA-loaded calcium phosphate nanoparticles appear to be an effective non-viral vector for gene delivery and functioned well for odontogenic differentiation through Bmp2 transfection.
authors.affiliation: Radboud University Nijmegen Medical Centre, Department of Periodontology and Biomaterials, Nijmegen, The Netherlands.
authors.forename: Xuechao
authors.initials: X
authors.lastname: Yang
authors.forename: X Frank
authors.initials: XF
authors.lastname: Walboomers
authors.forename: Juliette
authors.initials: J
authors.lastname: van den Dolder
authors.forename: Fang
authors.initials: F
authors.lastname: Yang
authors.forename: Zhuan
authors.initials: Z
authors.lastname: Bian
authors.forename: Mingwen
authors.initials: M
authors.lastname: Fan
authors.forename: John A
authors.initials: JA
authors.lastname: Jansen
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000818:Animals; D000954:Antigens, Surface / Q000096:biosynthesis; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000235:genetics; D002130:Calcium Phosphates; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation; D002478:Cells, Cultured; D003782:Dental Pulp / Q000378:metabolism* / Q000648:ultrastructure; D005786:Gene Expression Regulation; D006801:Humans; D053758:Nanoparticles; D009805:Odontogenesis / Q000235:genetics; D051381:Rats; D013234:Stem Cells / Q000378:metabolism* / Q000648:ultrastructure; D014025:Titanium; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000954:Antigens, Surface; C525718:BMP2 protein, human; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D002130:Calcium Phosphates; C527498:STRO-1 antigen, rat; D016212:Transforming Growth Factor beta; C020243:calcium phosphate; D014025:Titanium; D000469:Alkaline Phosphatase