pH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein

Dan Xu,1 Fei Wu,1 Yinghui Chen,2,* Liangming Wei,3,* Weien Yuan1,* 1School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 2Department of Neurology, Jinshan Hospital, Fudan University, Shanghai, 3Key Laboratory for Thin Film and Microfabrication of the Ministry of Education, Institute of Micro...

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Autores principales:	Xu D, Wu F, Chen Y, Wei L, Yuan W
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2013
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/1690dd5a53f44bb681fe71f2ee0c2552
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spelling	oai:doaj.org-article:1690dd5a53f44bb681fe71f2ee0c25522021-12-02T02:31:36ZpH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein1176-91141178-2013https://doaj.org/article/1690dd5a53f44bb681fe71f2ee0c25522013-09-01T00:00:00Zhttp://www.dovepress.com/ph-sensitive-degradable-nanoparticles-for-highly-efficient-intracellul-a14242https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Dan Xu,1 Fei Wu,1 Yinghui Chen,2,* Liangming Wei,3,* Weien Yuan1,* 1School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 2Department of Neurology, Jinshan Hospital, Fudan University, Shanghai, 3Key Laboratory for Thin Film and Microfabrication of the Ministry of Education, Institute of Micro/Nano Science and Technology, Shanghai Jiao Tong University, Shanghai, People's Republic of China*These authors contributed equally to this workBackground: Encapsulating exogenous proteins into a nanosized particulate system for delivery into cells is a great challenge. To address this issue, we developed a novel nanoparticle delivery method that differs from the nanoparticles reported to date because its core was composed of cross-linked dextran glassy nanoparticles which had pH in endosome-responsive environment and the protein was loaded in the core of cross-linked dextran glassy nanoparticles.Methods: In this study, dextran in a poly(ethylene glycol) aqueous two-phase system created a different chemical environment in which proteins were encapsulated very efficiently (84.3% and 89.6% for enhanced green fluorescent protein and bovine serum albumin, respectively) by thermodynamically favored partition. The structures of the nanoparticles were confirmed by confocal laser scanning microscopy and scanning electron microscopy.Results: The nanoparticles had a normal size distribution and a mean diameter of 186 nm. MTT assays showed that the nanoparticles were nontoxic up to a concentration of 2000 µg/mL in human hepatocarcinoma cell line SMMC-7721, HeLa, and BRL-3A cells. Of note, confocal laser scanning microscopy studies showed that nanoparticles loaded with fluorescein isothiocyanate-bovine serum albumin were efficiently delivered and released proteins into the cytoplasm of HeLa cells. Flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling assays showed that nanoparticles with a functional protein (apoptin) efficiently induced significant tumor cell apoptosis, which was confirmed by DAPI staining.Conclusion: Our findings indicate that these nanoparticles meet the high demands for delivering protein medicines and have great potential in protein therapy.Keywords: cellular uptake, protein delivery, nanoparticles, apoptosisXu DWu FChen YWei LYuan WDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2013, Iss default, Pp 3405-3414 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Xu D Wu F Chen Y Wei L Yuan W pH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein
description	Dan Xu,1 Fei Wu,1 Yinghui Chen,2,* Liangming Wei,3,* Weien Yuan1,* 1School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 2Department of Neurology, Jinshan Hospital, Fudan University, Shanghai, 3Key Laboratory for Thin Film and Microfabrication of the Ministry of Education, Institute of Micro/Nano Science and Technology, Shanghai Jiao Tong University, Shanghai, People's Republic of China*These authors contributed equally to this workBackground: Encapsulating exogenous proteins into a nanosized particulate system for delivery into cells is a great challenge. To address this issue, we developed a novel nanoparticle delivery method that differs from the nanoparticles reported to date because its core was composed of cross-linked dextran glassy nanoparticles which had pH in endosome-responsive environment and the protein was loaded in the core of cross-linked dextran glassy nanoparticles.Methods: In this study, dextran in a poly(ethylene glycol) aqueous two-phase system created a different chemical environment in which proteins were encapsulated very efficiently (84.3% and 89.6% for enhanced green fluorescent protein and bovine serum albumin, respectively) by thermodynamically favored partition. The structures of the nanoparticles were confirmed by confocal laser scanning microscopy and scanning electron microscopy.Results: The nanoparticles had a normal size distribution and a mean diameter of 186 nm. MTT assays showed that the nanoparticles were nontoxic up to a concentration of 2000 µg/mL in human hepatocarcinoma cell line SMMC-7721, HeLa, and BRL-3A cells. Of note, confocal laser scanning microscopy studies showed that nanoparticles loaded with fluorescein isothiocyanate-bovine serum albumin were efficiently delivered and released proteins into the cytoplasm of HeLa cells. Flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling assays showed that nanoparticles with a functional protein (apoptin) efficiently induced significant tumor cell apoptosis, which was confirmed by DAPI staining.Conclusion: Our findings indicate that these nanoparticles meet the high demands for delivering protein medicines and have great potential in protein therapy.Keywords: cellular uptake, protein delivery, nanoparticles, apoptosis
format	article
author	Xu D Wu F Chen Y Wei L Yuan W
author_facet	Xu D Wu F Chen Y Wei L Yuan W
author_sort	Xu D
title	pH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein
title_short	pH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein
title_full	pH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein
title_fullStr	pH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein
title_full_unstemmed	pH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein
title_sort	ph-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein
publisher	Dove Medical Press
publishDate	2013
url	https://doaj.org/article/1690dd5a53f44bb681fe71f2ee0c2552
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pH-sensitive degradable nanoparticles for highly efficient intracellular delivery of exogenous protein

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