A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system

The transport of DNA into eukaryotic cells is minimal because of the cell membrane barrier, and this limits the application of DNA vaccines, gene silencing, and gene therapy. Several available transfection reagents and techniques have been used to circumvent this problem. Alternatively, nonviral nan...

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Main Authors:	Eroglu, Erdal, Tiwari, Pooja M, Waffo, Alain B, Miller, Michael E, Vig, Komal, Dennis, Vida A, Singh, Shree R
Format:	Online
Language:	English
Published:	Dove Medical Press 2013
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3629879/

id	pubmed-3629879
recordtype	oai_dc
spelling	pubmed-36298792013-04-22 A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system Eroglu, Erdal Tiwari, Pooja M Waffo, Alain B Miller, Michael E Vig, Komal Dennis, Vida A Singh, Shree R Original Research The transport of DNA into eukaryotic cells is minimal because of the cell membrane barrier, and this limits the application of DNA vaccines, gene silencing, and gene therapy. Several available transfection reagents and techniques have been used to circumvent this problem. Alternatively, nonviral nanoscale vectors have been shown to bypass the eukaryotic cell membrane. In the present work, we developed a unique nanomaterial, pHEMA+chitosan nanospheres (PCNSs), which consisted of poly(2-hydroxyethyl methacrylate) nanospheres surrounded by a chitosan cationic shell, and we used this for encapsulation of a respiratory syncytial virus (RSV)-F gene construct (a model for a DNA vaccine). The new nanomaterial was capable of transfecting various eukaryotic cell lines without the use of a commercial transfection reagent. Using transmission electron microscopy, (TEM), fluorescence activated cell sorting (FACS), and immunofluorescence, we clearly demonstrated that the positively charged PCNSs were able to bind to the negatively charged cell membrane and were taken up by endocytosis, in Cos-7 cells. Using quantitative polymerase chain reaction (qPCR), we also evaluated the efficiency of transfection achieved with PCNSs and without the use of a liposomal-based transfection mediator, in Cos-7, HEp-2, and Vero cells. To assess the transfection efficiency of the PCNSs in vivo, these novel nanomaterials containing RSV-F gene were injected intramuscularly into BALB/c mice, resulting in high copy number of the transgene. In this study, we report, for the first time, the application of the PCNSs as a nanovehicle for gene delivery in vitro and in vivo. Dove Medical Press 2013 2013-04-11 /pmc/articles/PMC3629879/ /pubmed/23610520 http://dx.doi.org/10.2147/IJN.S43168 Text en © 2013 Eroglu et al, publisher and licensee Dove Medical Press Ltd. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Eroglu, Erdal Tiwari, Pooja M Waffo, Alain B Miller, Michael E Vig, Komal Dennis, Vida A Singh, Shree R
spellingShingle	Eroglu, Erdal Tiwari, Pooja M Waffo, Alain B Miller, Michael E Vig, Komal Dennis, Vida A Singh, Shree R A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system
author_facet	Eroglu, Erdal Tiwari, Pooja M Waffo, Alain B Miller, Michael E Vig, Komal Dennis, Vida A Singh, Shree R
author_sort	Eroglu, Erdal
title	A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system
title_short	A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system
title_full	A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system
title_fullStr	A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system
title_full_unstemmed	A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system
title_sort	nonviral phema+chitosan nanosphere-mediated high-efficiency gene delivery system
description	The transport of DNA into eukaryotic cells is minimal because of the cell membrane barrier, and this limits the application of DNA vaccines, gene silencing, and gene therapy. Several available transfection reagents and techniques have been used to circumvent this problem. Alternatively, nonviral nanoscale vectors have been shown to bypass the eukaryotic cell membrane. In the present work, we developed a unique nanomaterial, pHEMA+chitosan nanospheres (PCNSs), which consisted of poly(2-hydroxyethyl methacrylate) nanospheres surrounded by a chitosan cationic shell, and we used this for encapsulation of a respiratory syncytial virus (RSV)-F gene construct (a model for a DNA vaccine). The new nanomaterial was capable of transfecting various eukaryotic cell lines without the use of a commercial transfection reagent. Using transmission electron microscopy, (TEM), fluorescence activated cell sorting (FACS), and immunofluorescence, we clearly demonstrated that the positively charged PCNSs were able to bind to the negatively charged cell membrane and were taken up by endocytosis, in Cos-7 cells. Using quantitative polymerase chain reaction (qPCR), we also evaluated the efficiency of transfection achieved with PCNSs and without the use of a liposomal-based transfection mediator, in Cos-7, HEp-2, and Vero cells. To assess the transfection efficiency of the PCNSs in vivo, these novel nanomaterials containing RSV-F gene were injected intramuscularly into BALB/c mice, resulting in high copy number of the transgene. In this study, we report, for the first time, the application of the PCNSs as a nanovehicle for gene delivery in vitro and in vivo.
publisher	Dove Medical Press
publishDate	2013
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3629879/
_version_	1611971137084850176

A nonviral pHEMA+chitosan nanosphere-mediated high-efficiency gene delivery system

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