Homogeneous synthesis of quaternized chitin in NaOH/urea aqueous solution as a potential gene vector.
Water-soluble quaternized chitins (QCs) were homogeneously synthesized by reacting chitin with (3-chloro-2-hydroxypropyl) trimethylammonium chloride (CHPTAC) in 8wt% NaOH/4wt% urea aqueous solutions. The chemical structure and solution properties of the quaternized chitins were characterized by (1)H NMR, FT-IR, elemental analysis, dynamic light scattering (DLS) and zeta potential measurements. The results demonstrated that the water-soluble QCs, with a degree of substitution (DS) values of 0.27-0.54, could be obtained by varying the concentration of chitin, the molar ratio of CHPTAC to chitin unit, and the reaction time at room temperature (25°C). Two QCs (DS=0.36 and 0.54) were selected and studied as gene carriers. Agarose gel retardation assay revealed that both QCs could condense DNA efficiently when N/P ratio>3. The results of particle size and zeta potential indicated that both QCs had a good ability of condensing plasmid DNA into compact nanoparticles with the size of 100-200nm and zeta potential of +18 to +36mV. Compared to polyethylenimine (PEI, 25kDa), the QCs exhibited outstanding low cytotoxicity. Transfection efficiencies of the QCs/DNA complexes were measured using pGL-3 encoding luciferase as the foreign DNA, and the QCs/DNA complexes showed effective transfection efficiencies in 293T cells. These results revealed that the QCs prepared in NaOH/urea aqueous solutions could be used as promising non-viral gene carriers owing to their excellent characteristics.
Wuhan University, Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China; Guangzhou Sugarcane Industry Research Institute, Guangzhou 510316, China.
Na
N
Peng
Wuhan University, Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China.
Ziye
Z
Ai
Wuhan University, Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China.
Zehong
Z
Fang
Wuhan University, Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China.
Yanfeng
Y
Wang
Wuhan University, Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China.
Zhiping
Z
Xia
Wuhan University, Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China.
Zibiao
Z
Zhong
Wuhan University, Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China.
Xiaoli
X
Fan
Wuhan University, Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China; The 3rd Xiangya Hospital of Central South University, Research Center of National Health Ministry on Transplantation Medicine Engineering and Technology, Changsha 410013, China. Electronic address: yqf_china@163.com.
Qifa
Q
Ye
D060326:Chemistry Techniques, Synthetic; D002686:Chitin / Q000138:chemical synthesis* / Q000737:chemistry* / Q000633:toxicity; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004337:Drug Carriers / Q000138:chemical synthesis / Q000737:chemistry / Q000633:toxicity; D057809:HEK293 Cells; D006801:Humans; D020005:Propanols / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012972:Sodium Hydroxide / Q000737:chemistry*; D012996:Solutions; D013696:Temperature; D014162:Transfection / Q000379:methods*; D014508:Urea / Q000737:chemistry*; D014867:Water / Q000737:chemistry*
D004337:Drug Carriers; D020005:Propanols; D000644:Quaternary Ammonium Compounds; D012996:Solutions; D014867:Water; D002686:Chitin; D012972:Sodium Hydroxide; D014508:Urea; D004247:DNA; C496948:(3-chloro-2-hydroxypropyl)trimethylammonium chloride
Degree of substitution; Gene delivery; Homogeneous synthesis; Quaternized chitin