title: Preparation and evaluation of pH-responsive charge-convertible ternary complex FA-PEI-CCA/PEI/DNA with low cytotoxicity and efficient gene delivery.
abstract: Because the surface of the cationic polymer gene complex is positively charged, it can result in problems such as poor blood stability and cytotoxicity. Therefore, reducing the positive charge of the cationic gene complex without affecting its transfection efficiency is crucial. To achieve this objective, a pH-responsive charge-convertible ternary complex was developed in this study. Modified plyethylenimine (PEI) with two different degrees of substitution of NH2(plyethylenimine-1,2-cyclohexanedicarboxylic anhydride, PEI-CCA, and folic acid-plyethylenimine-1,2-cyclohexanedicarboxylic anhydride, FA-PEI-CCA) were first obtained by a chemical graft reaction. PEI-CCA and FA-PEI-CCA have significantly lower cytotoxicities and much better blood compatibilities than PEI does, and the former have an undifferentiated compression capability of DNA. The zeta potential values of the as-prepared ternary complexes (PEI-CCA/PEI/DNA and FA-PEI-CCA/PEI/DNA) were negative at pH 7.4 and positive at pH 6.5, with particle sizes of approximately 150nm. MTT assays demonstrated the significantly lower cytotoxicities of the ternary complexes compared to that of PEI/DNA. Moreover, the cytotoxicities of the ternary complexes were lower at pH 7.4 than pH 6.5. Transfection experiments in vitro revealed that the mean fluorescence intensities and transfection efficiencies of the ternary complexes were lower than for PEI/DNA at pH 7.4 but were almost the same at pH 6.5. The ternary complex with a FA group had significantly higher mean fluorescence intensity and transfection efficiency than did the ternary complex without it. In addition, the transfection experiment in 293T cells preliminarily validated the targeting function of the FA group of the ternary complex.
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China; Department of Pharmacy, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 201204, China.
authors.forename: Aijie
authors.initials: A
authors.lastname: Guo
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Shaohui
authors.initials: S
authors.lastname: Xu
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China. Electronic address: shenyuanyuan@sjtu.edu.cn.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Department of Anesthesiology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200233, China. Electronic address: cuishuning118@163.com.
authors.forename: Derong
authors.initials: D
authors.lastname: Cui
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China. Electronic address: srguo@sjtu.edu.cn.
authors.forename: Shengrong
authors.initials: S
authors.lastname: Guo
mesh_terms: D003509:Cyclohexanecarboxylic Acids / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D057809:HEK293 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007097:Imines / Q000737:chemistry*; D008954:Models, Biological; D011095:Polyethylenes / Q000737:chemistry*; D011108:Polymers / Q000009:adverse effects / Q000737:chemistry*; D014162:Transfection
chemical_list: D003509:Cyclohexanecarboxylic Acids; D007097:Imines; D011095:Polyethylenes; D011108:Polymers; C505405:poly(ethylene imine); D004247:DNA
keywords: 1, 2-cyclohexanedicarboxylic anhydride; Charge convertible polymer; Plyethylenimine; Ternary complex; Transfection efficiency; pH-responsive