Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles

Wen Wen Deng*, Xia Cao*, Miao Wang*, Rui Qu, Wei Yan Su, Yan Yang, Ya Wei Wei, Xi Ming Xu, Jiang Nan YuDepartment of Pharmaceutics, School of Pharmacy and Center for Nano Drug/Gene Delivery and Tissue Engineering, Jiangsu University, Zhenjiang, People’s Republic of China*These authors...

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Autores principales: Deng WW, Cao X, Wang M, Qu R, Su WY, Yang Y, Wei YW, Xu XM, Yu JN
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Publicado: Dove Medical Press 2012
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spelling oai:doaj.org-article:4ddcfa4c29e84f7c96ec688bccc06bce2021-12-02T02:49:00ZDelivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles1176-91141178-2013https://doaj.org/article/4ddcfa4c29e84f7c96ec688bccc06bce2012-03-01T00:00:00Zhttp://www.dovepress.com/delivery-of-a-transforming-growth-factor-beta-1-plasmid-to-mesenchymal-a9482https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Wen Wen Deng*, Xia Cao*, Miao Wang*, Rui Qu, Wei Yan Su, Yan Yang, Ya Wei Wei, Xi Ming Xu, Jiang Nan YuDepartment of Pharmaceutics, School of Pharmacy and Center for Nano Drug/Gene Delivery and Tissue Engineering, Jiangsu University, Zhenjiang, People’s Republic of China*These authors contributed equally to this workAbstract: 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 LipofectamineTM 2000 (P = 0.002, Student’st-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.Keywords: nonviral gene vector, transfection, plasmid DNA, spermineDeng WWCao XWang MQu RSu WYYang YWei YWXu XMYu JNDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2012, Iss default, Pp 1297-1311 (2012)
institution DOAJ
collection DOAJ
language EN
topic Medicine (General)
R5-920
spellingShingle Medicine (General)
R5-920
Deng WW
Cao X
Wang M
Qu R
Su WY
Yang Y
Wei YW
Xu XM
Yu JN
Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles
description Wen Wen Deng*, Xia Cao*, Miao Wang*, Rui Qu, Wei Yan Su, Yan Yang, Ya Wei Wei, Xi Ming Xu, Jiang Nan YuDepartment of Pharmaceutics, School of Pharmacy and Center for Nano Drug/Gene Delivery and Tissue Engineering, Jiangsu University, Zhenjiang, People’s Republic of China*These authors contributed equally to this workAbstract: 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 LipofectamineTM 2000 (P = 0.002, Student’st-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.Keywords: nonviral gene vector, transfection, plasmid DNA, spermine
format article
author Deng WW
Cao X
Wang M
Qu R
Su WY
Yang Y
Wei YW
Xu XM
Yu JN
author_facet Deng WW
Cao X
Wang M
Qu R
Su WY
Yang Y
Wei YW
Xu XM
Yu JN
author_sort Deng WW
title Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles
title_short Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles
title_full Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles
title_fullStr Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles
title_full_unstemmed Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles
title_sort delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized pleurotus eryngii polysaccharide nanoparticles
publisher Dove Medical Press
publishDate 2012
url https://doaj.org/article/4ddcfa4c29e84f7c96ec688bccc06bce
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