title: Effect of acyl chain length and unsaturation on physicochemical properties and transfection efficiency of N-acyl-substituted low-molecular-weight chitosan.
abstract: The effects of acyl chain length and unsaturation on physicochemical characteristics and transfection efficiency of novel nanomicelles of N-acyl-substituted low-molecular-weight chitosan (N-acyl LMWC) were studied. After transfection optimization, 18-carbon chain length grafts were selected, and N-acyl LMWCs were prepared with increasing unsaturation (18:1-18:3 carbon acyl grafts). N-acyl LMWCs were characterized using infrared spectroscopy and elemental analysis. The effect of DNA addition on size and zeta potential of N-acyl LMWCs was determined by dynamic light scattering. N-acyl LMWC-plasmid DNA (pDNA) polyplex stability was confirmed using gel electrophoresis. Transfection efficiency of the derivative polymers was visualized in human embryonic kidney cells using a plasmid encoding green fluorescent protein by confocal fluorescence microscopy and was quantified using therapeutic plasmids encoding for interleukin-4 and interleukin-10. N-acyl LMWCs could form cationic nanomicelles with average hydrodynamic size between 73 and 132 nm. DNA addition to nanomicelles led to minimal increase in the size. N-acyl LMWC-pDNA polyplexes showed excellent stability on storage and could protect DNA from enzymatic degradation. The transfection efficiencies of N-acyl LMWCs with 18:1 and 18:2 grafts were comparable with FuGENE® HD but were approximately eightfold and 35-fold greater as compared with LMWC and naked DNA, respectively.
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, Nursing, and Allied Sciences, North Dakota State University, Fargo, North Dakota 58102, USA.
authors.forename: Rhishikesh
authors.initials: R
authors.lastname: Mandke
authors.forename: Jagdish
authors.initials: J
authors.lastname: Singh
mesh_terms: D001672:Biocompatible Materials; D002461:Cell Line, Transformed; D048271:Chitosan / Q000008:administration & dosage / Q000009:adverse effects / Q000138:chemical synthesis* / Q000737:chemistry*; D004247:DNA / Q000235:genetics; D004355:Drug Stability; D004356:Drug Storage; D005822:Genetic Vectors / Q000008:administration & dosage / Q000009:adverse effects / Q000138:chemical synthesis* / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics; D015847:Interleukin-4 / Q000235:genetics; D008823:Micelles; D008970:Molecular Weight; D010316:Particle Size; D010957:Plasmids / Q000235:genetics; D011108:Polymers / Q000008:administration & dosage / Q000009:adverse effects / Q000138:chemical synthesis* / Q000737:chemistry*; D013055:Spectrophotometry, Infrared / Q000379:methods; D014162:Transfection / Q000379:methods
chemical_list: D001672:Biocompatible Materials; D008823:Micelles; D011108:Polymers; D016753:Interleukin-10; D015847:Interleukin-4; D004247:DNA; D048271:Chitosan