Chitosan nanoparticles for plasmid DNA delivery: effect of chitosan molecular structure on formulation and release characteristics.
ABSTRACT Chitosan can be useful as a nonviral vector for gene delivery. Although there are several reports to form chitosan-pDNA particles, the optimization and effect on transfection remain insufficient. The chitosan-pDNA nanoparticles were formulated using complex coacervation and solvent evaporation techniques. The important parameters for the encapsulation efficiency were investigated, including molecular weight and deacetylation degree of chitosan. We found that encapsulation efficiency of pDNA is directly proportional with deacetylation degree, but there is an inverse proportion with molecular weight of chitosan. DNA-nanoparticles in the size range of 450-820 nm depend on the formulation process. The surface charge of the nanoparticles prepared with complex coacervation method was slightly positive with a zeta potential of +9 to +18 mV; nevertheless, nanoparticles prepared with solvent evaporation method had a zeta potential approximately +30 mV. The pDNA-chitosan nanoparticles prepared by using high deacetylation degree chitosan having 92.7%, 98.0%, and 90.4% encapsulation efficiency protect the encapsulated pDNA from nuclease degradation as shown by electrophoretic mobility analysis. The release of pDNA from the formulation prepared by complex coacervation was completed in 24 hr whereas the formulation prepared by evaporation technique released pDNA in 96 hr, but these release profiles are not statistically significant compared with formulations with similar structure (p > .05). According to the results, we suggest nanoparticles have the potential to be used as a transfer vector in further studies.
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ABSTRACT: Exogenous nucleotide supplementation during times of rapid growth and stress is preferred because de novo synthesis is insufficient and energetically a costly process. To overcome inefficient utilization of dietary nucleotides due to intestinal cell repulsion and dependency on pH, an efficient controlled delivery system based on chitosan nanoparticles (NPs) was developed. The effects of these (0.2% and 0.4%) RNA-loaded chitosan NPs (Chitosan: RNA ratio 2:1), 0.4% bare RNA, and 0.8% chitosan NPs on productive efficiency (growth rate, feed conversion and protein efficiency ratio), body composition, organo-somatic indices, haemato-biochemical and immune responses (WBC count, phagocytic activity, serum lysozyme, serum total protein and albumin : globulin ratio) and survival of Labeo rohita fish fingerlings, following challenge with pathogenic bacteria were evaluated. Dietary chitosan NPs were found not to affect productive efficiency, but improved (P 0.05) by dietary treatments. RNA-loaded chitosan NPs increased hepato- and viscerosomatic indices. The activity of metabolic enzymes (intestinal and liver alkaline phosphatase, alanine- and aspartateamino transferases, lactate- and malate-dehydrogeneases) corresponded with the performance of the respective diets. As growth, immunity and disease resistance in fish given dietary nano-sized RNA were significantly higher than in those given bare RNA or chitosan alone, and as nanoformulation reduced the usage of individual components by half, the use of RNA-loaded chitosan NPs can be favoured in the feed/food industry over chitosan and RNA alone.Current Nanoscience 01/2014; 10(3). · 1.42 Impact Factor
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ABSTRACT: Chitosan nanoparticles (CS NPs) were prepared as a carrier for Human papillomavirus type 16 HPV-16) E7 gene and their gene transfection ability were evaluated in vitro. The plasmid expressing green fluorescent protein (pEGFP) was used as a reporter gene. Gel electrophoresis demonstrated full binding of CS NPs with the pDNA. The transfection of CS-pEGFP NPs was efficient in CHO cells and the expression of green fluorescent proteins was well observed. The expression of E7 proteins was confirmed under SDS-PAGE and western blot analysis. As a conclusion CS NPs may serve as an effective nonviral carrier for delivery of nucleotides into eukaryotic cells.Artificial Cells Blood Substitutes and Biotechnology 03/2014; · 0.71 Impact Factor
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ABSTRACT: Various N-acyl chitosans with propionyl-, hexanoyl-, nonanoyl-, lauroyl-, pentadecanoyl-, and stearoyl-groups were synthesized and self-aggregated N-acyl chitosan nanoparticles (CSNPs) were prepared by sonication. By the modification with N-acyl groups, CSNPs increased their hydrophobic character and changed its structural features to be more suitable as a delivery carrier. The mean diameters of bovine serum albumin (BSA)-loaded N-acyl CSNPs ranged from 138 to 551 nm. Uniform particle size distribution of BSA-loaded N-acyl CSNPs was observed. The protein loading efficiency of N-acyl CSNPs was about 94–95% with lower BSA concentration (0.1 mg/mL) and not significantly different with acyl chain length. With higher BSA concentration (1.0 mg/mL), however, the highest protein loading efficiency was observed with lauroyl and pentadecanoyl CSNPs. The results suggest that lauroyl and pentadecanoyl CSs are interesting candidates for protein delivery system. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012Journal of Applied Polymer Science 04/2012; 124(2). · 1.40 Impact Factor