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Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles.
- Source :
-
International journal of nanomedicine [Int J Nanomedicine] 2012; Vol. 7, pp. 1297-311. Date of Electronic Publication: 2012 Mar 14. - Publication Year :
- 2012
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Abstract
- The objective of this study was to investigate the use of cationized Pleurotus eryngii polysaccharide (CPEPS) as a nonviral gene delivery vehicle to transfer plasmid DNA encoding transforming growth factor beta-1 (pTGF-β1) into mesenchymal stem cells (MSCs) in vitro. Crude P. eryngii polysaccharide was purified, and then cationized by grafting spermine onto the backbone of the polysaccharide. Agarose gel electrophoresis, transmission electron microscopy, and a Nano Sense Zetasizer (Malvern Instruments, Malvern, UK) were used to characterize the CPEPS-pTGF-β1 nanoparticles. The findings of cytotoxicity analysis showed that when the nanoparticles were formulated with a CPEPS/pTGF-β1 weight ratio ≥ 10:1, a greater gel retardation effect was observed during agarose gel electrophoresis. The CPEPS-pTGF-β1 nanoparticles with a weight ratio of 20:1, respectively, possessed an average particle size of 80.8 nm in diameter and a zeta potential of +17.4 ± 0.1 mV. Significantly, these CPEPS-pTGF-β1 nanoparticles showed lower cytotoxicity and higher transfection efficiency than both polyethylenimine (25 kDa) (P = 0.006, Student's t-test) and Lipofectamine(TM) 2000 (P = 0.002, Student's t-test). Additionally, the messenger RNA expression level of TGF-β1 in MSCs transfected with CPEPS-pTGF-β1 nanoparticles was significantly higher than that of free plasmid DNA-transfected MSCs and slightly elevated compared with that of Lipofectamine 2000-transfected MSCs. Flow cytometry analysis demonstrated that 92.38% of MSCs were arrested in the G1 phase after being transfected with CPEPS-pTGF-β1 nanoparticles, indicating a tendency toward differentiation. In summary, the findings of this study suggest that the CPEPS-pTGF-β1 nanoparticles prepared in this work exhibited excellent transfection efficiency and low toxicity. Therefore, they could be developed into a promising nonviral vector for gene delivery in vitro.
- Subjects :
- Analysis of Variance
Cations chemistry
Cell Survival drug effects
DNA administration & dosage
DNA chemistry
DNA genetics
Flow Cytometry
Genetic Vectors genetics
Lipids
Particle Size
Polysaccharides chemistry
RNA, Messenger analysis
RNA, Messenger genetics
Spermine chemistry
Transforming Growth Factor beta1 metabolism
Mesenchymal Stem Cells physiology
Nanoparticles chemistry
Plasmids administration & dosage
Plasmids genetics
Pleurotus chemistry
Polysaccharides administration & dosage
Transfection methods
Transforming Growth Factor beta1 genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1178-2013
- Volume :
- 7
- Database :
- MEDLINE
- Journal :
- International journal of nanomedicine
- Publication Type :
- Academic Journal
- Accession number :
- 22457592
- Full Text :
- https://doi.org/10.2147/IJN.S28010