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. 2013 Apr 1;15(4):1651.
doi: 10.1007/s11051-013-1651-0.

The novel albumin-chitosan core-shell nanoparticles for gene delivery: preparation, optimization and cell uptake investigation

Mahdi Karimi  1 , Pinar Avci  2 , Rezvan Mobasseri  1 , Michael R Hamblin  2 , Hossein Naderi-Manesh  1
Affiliations

The novel albumin-chitosan core-shell nanoparticles for gene delivery: preparation, optimization and cell uptake investigation

Mahdi Karimi et al. J Nanopart Res. .

Abstract

Natural polymers and proteins such as chitosan (CS) and albumin (Alb) have recently attracted much attention both in drug delivery and gene delivery. The underlying rationale is their unique properties such as biodegradability, biocompatibility and controlled release. This study aimed to prepare novel albumin-chitosan-DNA (Alb-CS-DNA) core-shell nanoparticles as a plasmid delivery system and find the best conditions for their preparation. Phase separation method and ionic interaction were used for preparation of Alb nanoparticles and Alb-CS-DNA core-shell nanoparticles, respectively. The effects of three important independent variables (1) CS/Alb mass ratio, (2) the ratios of moles of the amine groups of cationic polymers to those of the phosphate groups of DNA (N/P ratio), and (3) Alb concentration, on the nanoparticle size and loading efficiency of the plasmid were investigated and optimized through Box-Behnken design of response surface methodology (RSM). The optimum conditions were found to be CS/Alb mass ratio = 3, N/P ratio = 8.24 and Alb concentration = 0.1 mg/mL. The most critical factors for the size of nanoparticles and loading efficiency were Alb concentration and N/P ratio. The optimized nanoparticles had an average size of 176 ± 3.4 nm and loading efficiency of 80 ± 3.9 %. Cytotoxicity experiments demonstrated that the prepared nanoparticles were not toxic. The high cellular uptake of nanoparticles (~85 %) was shown by flow cytometry and fluorescent microscopy.

Keywords: Albumin; Chitosan; Gene delivery; Nanoparticle; Optimization.

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Figures

Fig. 1
Fig. 1
Schematic representation of preparation of Alb-CS-DNA nanoparticles
Fig. 2
Fig. 2
SEM image of optimized Alb-CS-DNA nanoparticles
Fig. 3
Fig. 3
Response surface plots (a, b, c, d, e and f) showing the effect of CS/Alb mass ratio, N/P ratio and albumin concentration on the nanoparticles size and loading efficiency
Fig. 4
Fig. 4
FTIR of purified CS (a) and Alb nanoparticles (b) and Alb-CS-DNA nanoparticles (c)
Fig. 5
Fig. 5
MTT assay for different concentrations of the optimized Alb-CS-DNA nanoparticles, CS and Alb
Fig. 6
Fig. 6
Cellular uptake of Alb-CS nanoparticles, HeLa cells treated with nanoparticles for 4 h and the uptake assessed by flow cytometry
Fig. 7
Fig. 7
Cellular uptake study of FITC-labeled Alb-CS-DNA nanoparticles after 4 h of incubation a. Nucleus of cells has been identified through Hoechst staining b. Fluorescent image of HeLa cells c. (a) and (b) merged
See this image and copyright information in PMC

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