Transferrin-mediated PEGylated nanoparticles for delivery of DNA/PLL

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The purpose of this work was to determine the stability of pDNA/poly(L-lysine) complex(DNA/PLL) during microencapsulation, prepare transferrin (TF) conjugated PEGylatednanoparticles (TF-PEG-NP) loading DNA/PLL, and assess its physicochemicalcharacteristics and in vitro transfection efficiency. The DNA/PLL was prepared by mixingplasmid DNA (pDNA) in deionized water with various amounts of PLL. PEGylatednanoparticles (PEG-NP) loading DNA/PLL were prepared by a water–oil–water doubleemulsion solvent evaporation technique. TF-PEG-NP was prepared by coupling TFwith PEG-NP. The physicochemical characteristics of TF-PEG-NP and in vitrotransfection efficiency on K562 cells were measured. The results showed that free pDNAreserved its double supercoiled form (dsDNA) for only on average 25.7% aftersonification, but over 70% of dsDNA was reserved after pDNA was contracted withPLL. The particle size range of TF-PEG-NP loading DNA/PLL was 150–450 nmwith entrapment efficiency over 70%. TF-PEG-NP exhibited the low burst effect(<10%) within 1 day. After the first phase, DNA/PLL displayed a sustained release. The amountof cumulated DNA/PLL release from TF-PEG-NP with 2% polymer over 7 days was 85.4%for DNA/PLL (1:0.3 mass ratio), 59.8% and 43.1% for DNA/PLL (1:0.6) and DNA/PLL(1:1.0), respectively. To TF-PEG-NP loading DNA/PLL without chloroquine,the percentage of EGFP expressing cells was 28.9% for complexes consisting ofDNA/PLL (1:0.3), 38.5% and 39.7% for DNA/PLL (1:0.6) and DNA/PLL (1:1.0),respectively. In TF-PEG-NP loading DNA/PLL with chloroquine, more cellswere transfected, the percentage of positive cells was 37.6% (DNA/PLL, 1:0.3),47.1% (DNA/PLL, 1:0.6) and 45.8% (DNA/PLL, 1:1.0), which meant that thetransfection efficiency of pDNA was increased by over 50 times when PLL andTF-PEG-NP were jointly used as a plasmid DNA carrier, in particular, the maximalpercentage of positive cells (47.1%) from TF-PEG-NP loading DNA/PLL (1:0.6) wasabout 70 times the transfection efficiency of free plasmid DNA. The average cellviability of TF-PEG-NP loading DNA/PLL was about 90%, which meant thatTF-PEG-NP appeared to be safer than PLL alone. As a result, TF-PEG-NP loadingDNA/PLL could be a more effective non-viral vector for the delivery of pDNA.