title: Cross-linked polyethylenimine-tripolyphosphate nanoparticles for gene delivery.
abstract: The high transfection efficiency of polyethylenimine (PEI) makes it an attractive potential nonviral genetic vector for gene delivery and therapy. However, the highly positive charge of PEI leads to cytotoxicity and limits its application. To reduce the cytotoxicity of PEI, we prepared anion-enriched nanoparticles that combined PEI with tripolyphosphate (TPP). We then characterized the PEI-TPP nanoparticles in terms of size, zeta potential, and Fourier-transform infrared (FTIR) spectra, and assessed their transfection efficiency, cytotoxicity, and ability to resist deoxyribonuclease (DNase) I digestion. The cellular uptake of PEI-TPP with phosphorylated internal ribosome entry site-enhanced green fluorescent protein C1 or FAM (fluorouracil, Adriamycin [doxorubicin] and mitomycin)-labeled small interfering ribonucleic acids (siRNAs) was monitored by fluorescence microscopy and confocal laser microscopy. The efficiency of transfected delivery of plasmid deoxyribonucleic acid (DNA) and siRNA in vitro was 1.11- to 4.20-fold higher with the PEI-TPP particles (7.6% cross-linked) than with the PEI, at all N:P ratios (nitrogen in PEI to phosphorus in DNA) tested. The cell viability of different cell lines was more than 90% at the chosen N:P ratios of PEI-TPP/DNA complexes. Moreover, PEI-TPP nanoparticles resisted digestion by DNase I for more than 2 hours. The time-dependent absorption experiment showed that 7.6% of cross-linked PEI-TPP particles were internalized by 293T cells within 1 hour. In summary, PEI-TPP nanoparticles effectively transfected cells while conferring little or no toxicity, and thus have potential application in gene delivery.
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, People's Republic of China.
authors.forename: Xianzhang
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Laboratory Science, Guangdong Second Provincial Traditional Chinese Medicine Hospital, Guangzhou, People's Republic of China.
authors.forename: Sujing
authors.initials: S
authors.lastname: Shen
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, People's Republic of China.
authors.forename: Zhanfeng
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, People's Republic of China.
authors.forename: Junhua
authors.initials: J
authors.lastname: Zhuang
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005347:Fibroblasts; D057809:HEK293 Cells; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry / Q000633:toxicity*; D011122:Polyphosphates / Q000737:chemistry / Q000633:toxicity*; D034741:RNA, Small Interfering / Q000235:genetics; D011817:Rabbits; D014162:Transfection / Q000379:methods*
chemical_list: D011122:Polyphosphates; D034741:RNA, Small Interfering; D011094:Polyethyleneimine; C005692:triphosphoric acid
keywords: nanoparticles (NPs); polyethylenimine (PEI); transfection; tripolyphosphate (TPP)