Nanoparticles bearing polyethyleneglycol-coupled transferrin as gene carriers:: preparation and in vitro evaluation

The aims of this work were to determine the stability of pDNA against various conditions during microencapsulation, prepare transferrin (TF)-conjugated PEGylated polycyanoacrylate nanoparticles (TF-PEG-nanoparticles), and assess its physicochemical characteristics and in vitro targeting cells association. The open circular forms of pDNA obviously increased when pDNA was emulsified into organic solution under sonification. When pDNA solution (pH 7.0) contained 1, 3 or 5% (w/v) PVA, after sonification, average 48.2, 59.4 and 62.1% of double-supercoiled DNA (dsDNA) were preserved, respectively. When medium of pDNA was 0.9% NaCl (pH 7.0), 0.1 M NaHCO3 (pH 8.0) or phosphate buffer (pH 8.0), average 53.1, 69.3 and 56.9% of dsDNA remained after sonification, respectively. Poly(aminopoly(ethylene glycol)cyanoacrylate-co-hexadecyl cyanoacrylate) (poly(H(2)NPEGCA-co-HDCA)) showed a slight influence on pDNA in 0.1 M NaHCO3 (pH 8.0) when its concentration increased from 0.5 to 4% (w/v). TF-PEG-nanoparticles loading pDNA were spherical in shape with size under 200 nm and entrapment efficiency 35-50%. 0.1 M NaHCO3 with 3% PVA (w/v) could largely reduce the damage of pDNA during microencapsulation. TF-PEG-nanoparticles bore 1-3% of the total PEG chains conjugated to TF molecules, and exhibited the burst effect with over 30% drug release within I day. After the first phase, pDNA release profiles displayed a sustained release. The amount of cumulated pDNA release over 7 days was: 86.3, 81.5 and 74.4% for 1, 2 and 4% polymer nanoparticles, respectively. The degree of target K562 cell binding of TF-PEG-nanoparticles was greater than that of non-targeted PEG-nanoparticles at 4 degreesC. The presence of free TF decreased significantly the degree of cell binding of TF-PEG-nanoparticles, which revealed that the binding of TF-PEG-nanoparticles to K562 cells was indeed receptor specific. These results suggested that TF-PEG-nanoparticles were useful for delivery of pDNA to target cells. (C) 2003 Elsevier Science B.V. All rights reserved.