화학공학소재연구정보센터
홈 로그인 로그아웃 연락처 사이트맵
Macromolecular Research, Vol.23, No.7, 618-627, July, 2015
DOI10.1007/s13233-015-3074-5  Export Citation
Serum resistant and enhanced transfection of plasmid DNA by PEG-stabilized polyplex nanoparticles of L-histidine substituted polyethyleneimine
Haniye Najafi, Samira Sadat Abolmaali, Bahareh Owrangi, Younes Ghasemi1, and Ali Mohammad Tamaddon†
  • Center for Nanotechnology in Drug Delivery & Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Shiraz University of Medical Science, Shiraz, 71345, Iran
  • 1Pharmaceutical Sciences Research Center & Department of Pharmaceutical Biotechnology, Shiraz University of Medical Sciences, Shiraz, 71345, Iran
E-mail:
To improve transfection of plasmid DNA as well as serum protein stability of polyionic complex nanoparticles of branched polyethyleneimine (bPEI), poly(ethylene glycol) (PEG)-stabilized nanoparticles were made from L-histidine substituted bPEI (PEI-Histidine) synthesized by Fmoc chemistry. The polymer was characterized by TNBS assay, 1H NMR, GFC, potentiometric titration and elemental analysis of carbon and nitrogen, DNA condensation, and the stability against extracellular matrix (heparin sulfate) was investigated by dye exclusion and agarose gel retardation assays. The nanoparticles were characterized by dynamic light scattering-zeta potential analyzer. Cytotoxicity and expression of enhanced green fluorescent protein (EGFP) were determined in hepatocellular carcinoma by MTT assay and fluorescent techniques. PEI-Histidine showed a reduced pK a without any significant loss of total primary amines. Plasmid DNA was condensed almost thoroughly with PEGylated polymers, either bPEI or PEI-Histidine, at lower critical N/P ratios. PEGylated PEI-Histidine showed the better resistance to heparin induced displacement and the lower cytotoxicity when it was compared to bPEI. Interestingly unlike bPEI, smaller and less positively charged nanoparticles were obtained from PEGylated PEI-Histidine at N/P ratio=2 that resulted in about 4 folds higher EGFP expression than bPEI without any significant cytotoxicity. These properties are consistent with the higher serum protein resistance and buffer capacity of PEGylated PEI-Histidine at endosomal acidic pH.
Keywords:L-histidine substitution;PEGylated polyethyleneimine;serum resistance;transfection;plasmid DNA
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