Nano-Sized Calcium Phosphate Particles for Periodontal Gene Therapy.

Department of Periodontics, The University of Iowa College of Dentistry, Iowa City, IA.
Journal of Periodontology (Impact Factor: 2.57). 03/2012; DOI: 10.1902/jop.2012.120012
Source: PubMed

ABSTRACT Background: Growth factors such as platelet derived growth factor (PDGF) have significantly enhanced periodontal therapy outcomes but with a high degree of variability, mostly due to lack of their continual supply for a required period of time. One method to overcome this barrier is gene therapy and the aim of this in vitro study is to evaluate PDGF-B gene delivery in fibroblasts using nano-sized calcium phosphate particles (NCaPP) as vectors. Methods: NCaPP incorporating green fluorescent protein, GFP (NCaPP-GFP) and PDGF-B (NCaPP-PDGF-B) plasmids were synthesized using an established precipitation system and characterized using transmission electron microscopy and 1.2% agarose gel electrophoresis. Biocompatibility and transfection of the nanoplexes in fibroblasts were evaluated using cytotoxicity assay and florescence microscopy, respectively. Polymerase chain reaction (PCR) and enzyme linked Immunosorbent assay (ELISA) were performed to evaluate PDGF-B transfection after different time points of treatments and the functionality of PDGF-B transfection was evaluated using the cell proliferation assay. Results: Synthesized NCaPP nanoplexes incorporating the genes of GFP and PDGF-B were spherical in shape and measured about 30 to 50 nm in diameter. Gel electrophoresis confirmed DNA incorporation and stability within the nanoplexes and MTS assay demonstrated their biocompatibility in fibroblasts. In vitro transfection studies revealed a higher and longer lasting transfection after NCaPP-PDGF-B treatment, which lasted up to 96 hours. Significantly enhanced fibroblast proliferation observed in NCaPP-PDGF-B treated cells confirmed the functionality of these nanoplexes. Conclusion: NCaPP demonstrated higher levels of biocompatibility and efficiently transfected PDGF plasmids into fibroblasts under described in vitro conditions.

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