title: Nano-sized calcium phosphate particles for periodontal gene therapy.
abstract: BACKGROUND
  Growth factors such as platelet-derived growth factor (PDGF) have significantly enhanced periodontal therapy outcomes with a high degree of variability, mostly due to the lack of continual supply for a required period of time. One method to overcome this barrier is gene therapy. 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 (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 and enzyme-linked immunosorbent assay 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 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium reagent 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.
  
  
  CONCLUSIONS
  NCaPP demonstrated higher levels of biocompatibility and efficiently transfected PDGF plasmids into fibroblasts under described in vitro conditions.
authors.affiliation: Department of Periodontics, The University of Iowa College of Dentistry, Iowa City, IA 52242, USA. satheesh-elangovan@uiowa.edu
authors.forename: Satheesh
authors.initials: S
authors.lastname: Elangovan
authors.forename: Shardool
authors.initials: S
authors.lastname: Jain
authors.forename: Pei-Chin
authors.initials: PC
authors.lastname: Tsai
authors.forename: Henry C
authors.initials: HC
authors.lastname: Margolis
authors.forename: Mansoor
authors.initials: M
authors.lastname: Amiji
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000138:chemical synthesis; D002130:Calcium Phosphates / Q000138:chemical synthesis / Q000737:chemistry; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival; D005786:Gene Expression Regulation / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000138:chemical synthesis; D049452:Green Fluorescent Proteins; D049408:Luminescent Agents; D051379:Mice; D041681:NIH 3T3 Cells; D053758:Nanoparticles / Q000737:chemistry; D010510:Periodontal Diseases / Q000628:therapy*; D010957:Plasmids / Q000138:chemical synthesis; D020574:Proto-Oncogene Proteins c-sis / Q000235:genetics*; D013778:Tetrazolium Salts; D013844:Thiazoles; D014162:Transfection / Q000379:methods
chemical_list: D001672:Biocompatible Materials; D002130:Calcium Phosphates; D049408:Luminescent Agents; D020574:Proto-Oncogene Proteins c-sis; D013778:Tetrazolium Salts; D013844:Thiazoles; C070380:3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; D049452:Green Fluorescent Proteins