A comparative evaluation of poly-L-lysine-palmitic acid and Lipofectamine 2000 for plasmid delivery to bone marrow stromal cells.
The current study compared the effectiveness of an amphiphilic biomaterial poly(L-lysine)-palmitic acid (PLL-PA), and the lipid-based transfection agent Lipofectamine 2000 for plasmid delivery to bone marrow stromal cells (BMSC). We investigated the utility of the carriers to deliver a plasmid containing enhanced green fluorescent protein (pEGFP) to BMSC in vitro. Confocal microscopy was used to investigate the intracellular trafficking of pEGFP/carrier complexes. pEGFP delivery and EGFP expression were assessed by flow cytometry. PLL-PA formed condensed structures with pEGFP and successfully delivered the plasmid into the nucleus within 5 h of incubation with the cells. PLL-PA delivered the pEGFP to approximately 80% of the cells, achieving a maximum transfection efficiency of approximately 22%. This was significantly higher than Lipofectamine 2000-mediated transfection, which was 11% under most optimal conditions. Dosing the BMSC two or three times during the 24 h period increased the transfection efficiency by 2-3 folds, without compromising cell viability. When chloroquine was employed as an ensomolytic agent, 100 microM of the drug increased the transfection efficiency while reducing cell viability, but lower concentrations (1-10 microM) were not beneficial for transfection. Combining PLL-PA with Lipofectamine 2000 created an additive effect, increasing the transfection efficiency of PLL-PA. Long-term evaluation of gene expression with pEGFP/PLL-PA yielded approximately 17% transfection on day 1, which gradually decreased over a 12-day period. We conclude that PLL-PA is an effective biomaterial carrier and a promising candidate for non-viral gene delivery to BMSC.
Clements
Başak Açan
BA
Department of Chemical and Materials Engineering, Faculty of Engineering, University of Alberta, Edmonton, Alberta, Canada T6G 2G6.
Incani
Vanessa
V
Kucharski
Cezary
C
Lavasanifar
Afsaneh
A
Ritchie
Bruce
B
Uludağ
Hasan
H
D000818:Animals; D002478:Cells, Cultured; D004337:Drug Carriers / Q000737:chemistry*; D005260:Female; D008055:Lipids / Q000737:chemistry*; D008422:Materials Testing; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D019308:Palmitic Acid / Q000737:chemistry*; D010957:Plasmids / Q000008:administration & dosage*; D011107:Polylysine / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods*
D004337:Drug Carriers; D008055:Lipids; C086724:Lipofectamine; D011107:Polylysine; D019308:Palmitic Acid