Non-Viral Bone Morphogenetic Protein 2 Transfection of Rat Dental Pulp Stem Cells Using Calcium Phosphate Nanoparticles as Carriers

• Published in: Tissue engineering. Part A 2008-01, Vol.14 (1), p.71-81
• Main Authors: Yang, Xuechao, Walboomers, X. Frank, van den Dolder, Juliette, Yang, Fang, Bian, Zhuan, Fan, Mingwen, Jansen, John A.
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - biosynthesis; Animals; Antigens, Surface - biosynthesis; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins - biosynthesis; Bone Morphogenetic Proteins - genetics; Calcium Phosphates; Cell Differentiation - genetics; Cell Proliferation; Cells, Cultured; Dental Pulp - metabolism; Dental Pulp - ultrastructure; Gene Expression Regulation; Gene therapy; Genetic aspects; Humans; Methods; Nanoparticles; Odontogenesis - genetics; Proteins; Rats; Stem cells; Stem Cells - metabolism; Stem Cells - ultrastructure; Tissue engineering; Titanium; Transfection; Transforming Growth Factor beta - biosynthesis; Transforming Growth Factor beta - genetics
• ISSN: 1937-3341; 1937-335X
• DOI: 10.1089/ten.a.2007.0102

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.