N-alkylated chitosan as a potential nonviral vector for gene transfection.
Alkylated chitosans (ACSs) were prepared by modifying chitosan (CS) with alkyl bromide. The self-aggregation of ACSs in acetic acid solution was characterized by fluorescence spectroscopy and dynamic light scattering method. The results indicate that introducing alkyl side chains leads to the self-aggregation of ACSs, and CS with a 99% deacetylation degree shows no aggregation due to the electrostatic repulsion. The electrophoresis experiment demonstrates that the complex between CS and DNA was formed at a charge ratio (+/-) of 1/1; ACS/DNA complexes were formed at a lower charge ratio (+/-) of 1/4. A small amount of alkylated chitosans play the same shielding role as chitosan in protecting DNA from DNase hydrolysis. Differential scanning calorimetry (DSC) and atomic force microscopy (AFM) were employed separately to investigate the thermodynamic behavior of dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)/CS and DPPC/ACS mixtures and the variation in topological structure of DPPC membrane induced by CS and ACS. It is shown that CS and ACS can cause the fusion of DPPC multilamellar vesicles as well as membrane destabilization. In contrast, the perturbation effect induced by ACS is more evident due to the hydrophobic interaction. CS and ACS were used to transfer plasmid-encoding CAT into C(2)C(12) cell lines. Upon elongating the alkyl side chain, the transfection efficiency is increased and levels off after the number of carbons in the side chain exceeds 8. It is proposed that the higher transfection efficiency of ACS is attributed to the increasing entry into cells facilitated by hydrophobic interactions and easier unpacking of DNA from ACS carriers due to the weakening of electrostatic attractions between DNA and ACS.
Liu
Wen Guang
WG
Research Institute of Polymeric Materials, Tianjin University, Tianjin 300072.
Zhang
Xin
X
Sun
Shu Jun
SJ
Sun
Guang Jie
GJ
Yao
Kang De
KD
Liang
Dong Chun
DC
Guo
Gang
G
Zhang
Jing Yu
JY
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D000478:Alkylation; D000818:Animals; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002460:Cell Line; D002686:Chitin / Q000737:chemistry* / Q000494:pharmacology; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003851:Deoxyribonucleases / Q000737:chemistry / Q000378:metabolism; D005822:Genetic Vectors / Q000737:chemistry*; D008027:Light; D051379:Mice; D018625:Microscopy, Atomic Force / Q000379:methods; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence / Q000379:methods; D013816:Thermodynamics; D013997:Time Factors; D014162:Transfection / Q000379:methods*
D002686:Chitin; D015060:1,2-Dipalmitoylphosphatidylcholine; D004247:DNA; D003851:Deoxyribonucleases