title: Effect of N-pyridinium positions of quaternized chitosan on transfection efficiency in gene delivery system.
abstract: Methylated N-pyridylmethyl chitosan chlorides (M-PyMeChCs) with a similar total degree of quaternization (DQT) and molecular weight but different N-pyridinium positions were synthesized by reductive amination and methylation, respectively. The effect of N-pyridinium positions on transfection efficiency and cytotoxicity was investigated in human hepatoma (Huh7) cell lines. The results revealed that M-PyMeChCs are able to form a complete complex formation with DNA since there is an N/P ratio of 5. The particle sizes of M-PyMeChCs/DNA nanopolyplexes were approximately 300 nm and indicated a positive charge. The morphology of these nanopolyplexes was found to be in a spherical shape which was investigated by using the transmission electron microscopy (TEM). The M4-PyMeChC/DNA nanopolyplexes showed highest in vitro transfection efficiency in Huh7 cells at N/P ratio of 20 compared to M2-PyMeChC/DNA and M3-PyMeChC/DNA nanopolyplexes. In comparison to M3-PyMeChC/DNA nanopolyplexes, M2-PyMeChC/DNA and M4-PyMeChC/DNA nanopolyplexes showed lower cytotoxicity in Huh7 cells. Our result demonstrated that N-pyridinium positions of M-PyMeChCs are related to transfection efficiency and cytotoxicity.
authors.affiliation: Nanodelivery System Laboratory, National Nanotechnology Center, National Science and Technology Development Agency, Pathum Thani, Thailand. Electronic address: warayuth@nanotec.or.th.
authors.forename: Warayuth
authors.initials: W
authors.lastname: Sajomsang
authors.affiliation: Nanodelivery System Laboratory, National Nanotechnology Center, National Science and Technology Development Agency, Pathum Thani, Thailand.
authors.forename: Pattarapond
authors.initials: P
authors.lastname: Gonil
authors.affiliation: Nanodelivery System Laboratory, National Nanotechnology Center, National Science and Technology Development Agency, Pathum Thani, Thailand.
authors.forename: Uracha Rungsardthong
authors.initials: UR
authors.lastname: Ruktanonchai
authors.affiliation: Pharmaceutical Development of Green Innovations Group (PDGIG), Faculty of Pharmacy, Silpakorn University, Nakhon Pathom, Thailand.
authors.forename: Maleenart
authors.initials: M
authors.lastname: Petchsangsai
authors.affiliation: Pharmaceutical Development of Green Innovations Group (PDGIG), Faculty of Pharmacy, Silpakorn University, Nakhon Pathom, Thailand.
authors.forename: Praneet
authors.initials: P
authors.lastname: Opanasopit
authors.affiliation: Department of Manufacturing Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok, Thailand.
authors.forename: Satit
authors.initials: S
authors.lastname: Puttipipatkhachorn
mesh_terms: D045744:Cell Line, Tumor; D048271:Chitosan / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: C534744:methylated N-(4-pyridinylmethyl)chitosan; D049452:Green Fluorescent Proteins; D048271:Chitosan
keywords: Chitosan; Gene delivery; N-pyridylmethyl chitosan; Quaternized chitosan; Transfection efficiency