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In vitro and in vivo assessment of chitosan modified urocanic acid as gene carrier

YS. Hsueh, S. Subramaniam, YC. Tseng, TM. Chiang, O. Mestak, TK. Cheng, TF. Kuo, S. Sivasubramanian, FH. Lin, MJ. Shieh,

. 2017 ; 70 (Pt 1) : 599-606. [pub] 20160913

Jazyk angličtina Země Nizozemsko

Typ dokumentu časopisecké články

Perzistentní odkaz   https://www.medvik.cz/link/bmc17023594

Chitosan nanoparticles modified with 10 and 30% urocanic acid (CUA) via carbodiimide crosslinking were examined for an efficient gene delivery carrier. The CUA gene carrier was characterized by FTIR, TEM, SEM and the in vitro transfection efficiency CUA polyplex was tested with HeLa and 3T3 cells. The loading efficiency of CUA complexes with DNA was assessed at different N/P ratio of 1, 2, 4, 6, 8, and 10. The DNA loading efficiency was found be to >85% for chitosan, CUA10 and CUA30% and the DNA protection ability of CUA10 and CUA30 nanoparticle complexes was confirmed upon incubation with NheI and HindIII. The cell toxicity and cell viability results have supported the non-toxic nature of CUA10 and CUA30 nanoparticles. In vitro transfection efficiency of CUA10 and CUA30 polyplex was tested for EGFP expression in 3T3 and HeLa cells and a relative maximum % transfection of about 10% was confirmed by CUA10 and CUA30 after 96h transfection. The feasibility and biocompatibility of CUA gene carrier in transgenic chickens was also demonstrated. The in vitro transfection and in vivo embryonic viability studies further confirmed the CUA as promising gene carrier because of the improved biocompatibility and DNA protection ability.

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$a Chitosan nanoparticles modified with 10 and 30% urocanic acid (CUA) via carbodiimide crosslinking were examined for an efficient gene delivery carrier. The CUA gene carrier was characterized by FTIR, TEM, SEM and the in vitro transfection efficiency CUA polyplex was tested with HeLa and 3T3 cells. The loading efficiency of CUA complexes with DNA was assessed at different N/P ratio of 1, 2, 4, 6, 8, and 10. The DNA loading efficiency was found be to >85% for chitosan, CUA10 and CUA30% and the DNA protection ability of CUA10 and CUA30 nanoparticle complexes was confirmed upon incubation with NheI and HindIII. The cell toxicity and cell viability results have supported the non-toxic nature of CUA10 and CUA30 nanoparticles. In vitro transfection efficiency of CUA10 and CUA30 polyplex was tested for EGFP expression in 3T3 and HeLa cells and a relative maximum % transfection of about 10% was confirmed by CUA10 and CUA30 after 96h transfection. The feasibility and biocompatibility of CUA gene carrier in transgenic chickens was also demonstrated. The in vitro transfection and in vivo embryonic viability studies further confirmed the CUA as promising gene carrier because of the improved biocompatibility and DNA protection ability.
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$a Subramaniam, Sadhasivam $u Department of Microbial Biotechnology, Bharathiar University, Coimbatore, India.
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$a Tseng, Yi-Cheng $u Institute of Biomedical Engineering, National Taiwan University, No. 1, Sec. 1, Jen-Ai Road, Taipei, Taiwan.
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$a Chiang, Te-Ming $u Hospital and College of Medicine, National Taiwan University, Taipei, Taiwan.
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$a Mestak, Ondrej $u Department of Plastic Surgery, First Faculty of Medicine, Charles University in Prague, Bulovka Hospital, Budinova, Prague, Czech Republic.
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$a Cheng, Teng-Kuei $u Department of Animal Science and Technology, National Taiwan University, Taipei, Taiwan.
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$a Kuo, Tzong-Fu $u Institute of Veterinary Medicine, College of Veterinary Medicine, National Taiwan University, Taipei, Taiwan.
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$a Sivasubramanian, Savitha $u Department of Microbial Biotechnology, Bharathiar University, Coimbatore, India.
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$a Lin, Feng-Huei $u Institute of Biomedical Engineering, National Taiwan University, No. 1, Sec. 1, Jen-Ai Road, Taipei, Taiwan. Electronic address: double@ntu.edu.tw.
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