title: In vitro and in vivo assessment of chitosan modified urocanic acid as gene carrier.
abstract: 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.
authors: Institute of Biomedical Engineering, National Taiwan University, No. 1, Sec. 1, Jen-Ai Road, Taipei, Taiwan.
authors: Yu-Sheng
authors: YS
authors: Hsueh
authors: Department of Microbial Biotechnology, Bharathiar University, Coimbatore, India.
authors: Sadhasivam
authors: S
authors: Subramaniam
authors: Institute of Biomedical Engineering, National Taiwan University, No. 1, Sec. 1, Jen-Ai Road, Taipei, Taiwan.
authors: Yi-Cheng
authors: YC
authors: Tseng
authors: Hospital and College of Medicine, National Taiwan University, Taipei, Taiwan.
authors: Te-Ming
authors: TM
authors: Chiang
authors: Department of Plastic Surgery, First Faculty of Medicine, Charles University in Prague, Bulovka Hospital, Budinova, Prague, Czech Republic.
authors: Ondrej
authors: O
authors: Mestak
authors: Department of Animal Science and Technology, National Taiwan University, Taipei, Taiwan.
authors: Teng-Kuei
authors: TK
authors: Cheng
authors: Institute of Veterinary Medicine, College of Veterinary Medicine, National Taiwan University, Taipei, Taiwan.
authors: Tzong-Fu
authors: TF
authors: Kuo
authors: Department of Microbial Biotechnology, Bharathiar University, Coimbatore, India.
authors: Savitha
authors: S
authors: Sivasubramanian
authors: Institute of Biomedical Engineering, National Taiwan University, No. 1, Sec. 1, Jen-Ai Road, Taipei, Taiwan. Electronic address: double@ntu.edu.tw.
authors: Feng-Huei
authors: FH
authors: Lin
authors: Institute of Biomedical Engineering, National Taiwan University, No. 1, Sec. 1, Jen-Ai Road, Taipei, Taiwan.
authors: Ming-Jium
authors: MJ
authors: Shieh
mesh_terms: D016475:3T3 Cells; D000818:Animals; D030801:Animals, Genetically Modified; D016923:Cell Death; D002470:Cell Survival; D002642:Chick Embryo; D048271:Chitosan / Q000737:chemistry*; D004247:DNA / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry; D009555:Ninhydrin / Q000737:chemistry; D010316:Particle Size; D010957:Plasmids / Q000378:metabolism; D015183:Restriction Mapping; D017550:Spectroscopy, Fourier Transform Infrared; D055672:Static Electricity; D014162:Transfection; D014560:Urocanic Acid / Q000737:chemistry*
chemical_list: D004247:DNA; D048271:Chitosan; D004720:Endonucleases; D014560:Urocanic Acid; D009555:Ninhydrin
keywords: Chitosan; Gene carrier; Gene delivery; Transgenic chicken; Urocanic acid