Journal article
Characterization and evaluation of the efficacy of cationic complex mediated plasmid DNA delivery in human embryonic palatal mesenchyme cells
Journal of tissue engineering and regenerative medicine, Vol.10(11), pp.927-937
11/2016
DOI: 10.1002/term.1873
PMCID: PMC4425614
PMID: 24493354
Abstract
The purpose of this study was to develop and test a non-viral gene delivery system that can be employed to deliver genes of interest into a pre-osteoblastic cell line. Human embryonic palatal mesenchymal (HEPM 1486) cells were transfected with vector-plasmid DNA (pDNA) complexes. We explored calcium phosphate and polyethylenimine (PEI) as non-viral vectors and compared their respective in vitro transfection efficacies. Plasmid DNA encoding luciferase protein (LUC) was complexed with PEI (with differing N:P ratios) and calcium phosphate (with differing Ca:P ratios), using established protocols. The complexes prepared were then characterized for size and surface charge, using a Malvern Zetasizer Nano-ZS. The transfection efficiency and cytotoxicity of the prepared complexes were evaluated in HEPM cells. The PEI-pDNA complexes over the whole range of N:P ratios were found to be < 160 nm in size, while the calcium phosphate-pDNA complexes were relatively bigger. The PEI-pDNA complexes prepared at a N:P ratio of 10 were found to have maximum transfection efficiency at 4 h of treatment, with minimal cytotoxicity. The highest transfection efficiency obtained with calcium phosphate-pDNA complexes (Ca:P 200) was nearly 12-fold lower than that obtained with PEI-pDNA complexes (N:P 10). Following this, transgene expression in the HEPM cells treated with complexes prepared at a N:P ratio of 10 was further examined, using pDNA coding for enhanced green fluorescent protein (EGFP-N1) or therapeutically relevant platelet-derived growth factor B (PDGF-B). In conclusion, PEI was a more effective vector for delivering genes of interest to pre-osteoblasts than calcium phosphate. Copyright © 2014 John Wiley & Sons, Ltd.
Details
- Title: Subtitle
- Characterization and evaluation of the efficacy of cationic complex mediated plasmid DNA delivery in human embryonic palatal mesenchyme cells
- Creators
- Sheetal D'Mello - Division of Pharmaceutics and Translational Therapeutics, College of Pharmacy, University of Iowa, IA, USAAliasger K Salem - Division of Pharmaceutics and Translational Therapeutics, College of Pharmacy, University of Iowa, IA, USALiu Hong - Department of Periodontics, College of Dentistry, University of Iowa, IA, USASatheesh Elangovan - Department of Periodontics, College of Dentistry, University of Iowa, IA, USA
- Resource Type
- Journal article
- Publication Details
- Journal of tissue engineering and regenerative medicine, Vol.10(11), pp.927-937
- DOI
- 10.1002/term.1873
- PMID
- 24493354
- PMCID
- PMC4425614
- NLM abbreviation
- J Tissue Eng Regen Med
- ISSN
- 1932-7005
- eISSN
- 1932-7005
- Publisher
- England
- Grant note
- P30 CA086862 / NCI NIH HHS R21 CA133456 / NCI NIH HHS R21 CA128414 / NCI NIH HHS
- Language
- English
- Date published
- 11/2016
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Pharmaceutical Sciences and Experimental Therapeutics; Prosthodontics; Craniofacial Anomalies Research Center; Dental Research; Chemical and Biochemical Engineering; Periodontics
- Record Identifier
- 9983985850002771
Metrics
29 Record Views