Development of a calcium phosphate co-precipitate/poly(lactide-co-glycolide) DNA delivery system: release kinetics and cellular transfection studies.
One of the most common non-viral methods for the introduction of foreign deoxyribonucleic acid (DNA) into cultured cells is calcium phosphate co-precipitate transfection. This technique involves the encapsulation of DNA within a calcium phosphate co-precipitate, particulate addition to in vitro cell culture, endocytosis of the co-precipitate, and exogenous DNA expression by the transfected cell. In this study, we fabricated a novel non-viral gene transfer system by adsorbing DNA, encapsulated in calcium phosphate (DNA/Ca-P) co-precipitates, to biodegradable two- and three-dimensional poly(lactide-co-glycolide) matrices (2D-DNA/Ca-P/PLAGA, 3D-DNA/Ca-P/PLAGA). Co-precipitate release studies demonstrated an initial burst release over the first 48 h. By day 7, approximately 96% of the initially adsorbed DNA/Ca-P co-precipitate had been released. This was followed by low levels of co-precipitate release for 42 days. Polymerase chain reaction was used to demonstrate the ability of the released DNA containing co-precipitates to transfect SaOS-2 cells cultured in vitro on the 3D-DNA/Ca-P/PLAGA matrix and maintenance of the structural integrity of the exogenous DNA. In summary, a promising system for the incorporation and controlled delivery of exogenous genes encapsulated within a calcium phosphate co-precipitate from biodegradable polymeric matrices has been developed and may have applicability to the delivery of therapeutic genes and the transfection of other cell types.
Kofron
Michelle D
MD
Department of Biomedical Engineering, University of Virginia, Building MR-S, 415 Lane Road, Charlottesville, VA 22903, USA.
Laurencin
Cato T
CT
D000327:Adsorption; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D001483:Base Sequence; D002130:Calcium Phosphates / Q000737:chemistry*; D011232:Chemical Precipitation; D017931:DNA Primers; D004274:DNA, Recombinant / Q000008:administration & dosage*; D007700:Kinetics; D014162:Transfection
D023181:Antimicrobial Cationic Peptides; D002130:Calcium Phosphates; D017931:DNA Primers; D004274:DNA, Recombinant; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C045866:peptide-Gly-Leu-amide; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; C485829:calcium phosphate, dibasic, anhydrous