Title: Synthesis and characterization of thiolated chitosan-pDNA nanoparticles as gene carriers
Abstract: Objective: To study the preparation of thiolated chitosan and thiolated chitosan-plasmid DNA(pDNA)nanoparticles.Methods:Thiolated chitosan was prepared in the synthesis reaction of chitosan and thioglycolic acid with amide linkage forming by the catalysis of 1-thyl-3-(-dimethylaminopropyl)-carbodoomide hydrochloride(EDAC).The degree of thiol groups modification of the thiolated chitosan conjugate was determined by 5,5′-dithiobis(2-nitrobenzoiol acid)(DTNB) assay.When the plasmid pcDNA3.1(+)-EGFP was used as the reporter gene,thiolated chitosan-pcDNA3.1(+)-EGFP nanoparticles were prepared using a complex coacervation process by mixing chitosan and pDNA.The shape and size of thiolated chitosan-pcDNA3.1(+)-EGFP nanoparticles were observed by transmission electron microscope(TEM),and encapsulation rate was measured by ultraviolet spectrophotometer.The nanoparticles were treated by DNase I,then the products were measured by gel electrophoresis.Results:Chitosan was thiolatized by the catalysis of EDAC.1 gram of thiolated chitosan processed(202.85±3.05)μmol thiol groups(n=6)by calculating.It was demonstrated that full binding of thiolated chitosan with the pDNA and stability of nanoparticles.Encapsulation efficiency was higher than 90%.TEM images showed that nanoparticles were approximatively spherical in shape and particle size was in the range from 300 to 350 nm.The encapsulated pDNA protection against DNase I degradation was confirmed by DNase I treatment and gel electrophoresis.Conclusion:Thiolated chitosan-pcDNA3.1(+)-EGFP nanoparticles constructed are characterized for their proper particle size,high encapsulation efficiency of pDNA and ability protect pDNA from degradation.So Thiolated chitosan may serve as an effective nonviral gene carrier.
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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