Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin

Ehsan Hashemi,1,2 Hossein Mahdavi,3 Jafar Khezri,1 Farideh Razi,2,4 Mehdi Shamsara,1 Abbas Farmany5 1National Research Center for Transgenic Mouse & Animal Biotechnology Division, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran; 2Endocrinology and Metabolism Researc...

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Autores principales:	Hashemi E, Mahdavi H, Khezri J, Razi F, Shamsara M, Farmany A
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2019
Materias:	gene expression calcium doped magnetic nanograins cytotoxicity Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/f2445cc6cecf49a3b669d44f00551eed
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spelling	oai:doaj.org-article:f2445cc6cecf49a3b669d44f00551eed2021-12-02T08:51:56ZEnhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin1178-2013https://doaj.org/article/f2445cc6cecf49a3b669d44f00551eed2019-12-01T00:00:00Zhttps://www.dovepress.com/enhanced-gene-delivery-in-bacterial-and-mammalian-cells-using-pegylate-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Ehsan Hashemi,1,2 Hossein Mahdavi,3 Jafar Khezri,1 Farideh Razi,2,4 Mehdi Shamsara,1 Abbas Farmany5 1National Research Center for Transgenic Mouse & Animal Biotechnology Division, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran; 2Endocrinology and Metabolism Research Center, Endocrinology and Metabolism Clinical Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran; 3School of Chemistry, College of Science, University of Tehran, Tehran, Iran; 4Diabetes Research Center, Endocrinology and Metabolism Clinical Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran; 5Dental Implant Research Center, Hamadan University of Medical Sciences, Hamadan, IranCorrespondence: Abbas FarmanyDental Implant Research Center, Hamadan University of Medical Sciences, Hamadan, IranEmail a.farmany@ut.ac.irBackground: Beyond viral carriers which have been widely used in gene delivery, non-viral carriers can further improve the delivery process. However, the high cytotoxicity and low efficiency impedes the clinical application of non-viral systems. Therefore, in this work, we fabricated polyethylene glycol (PEG) coated, calcium doped magnetic nanograin (PEG/Ca(II)/Fe3O4) as a genome expression enhancer.Methods: Monodisperse magnetic nanograins (MNGs) with tunable size were synthesized by a solvothermal method. The citrate anions on the spherical surface of MNGs capture Ca2+ ions by an ion exchange process, which was followed by surface capping with PEG. The synthesized PEG/Ca(II)/Fe3O4 was characterized using Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) spectra, vibrating sample magnetometer (VSM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). MTT test was utilized to assess the toxicity of PEG/Ca(II)/Fe3O4. Real time qPCR was applied for quantification of gene expression.Results: DLS spectra and TEM images confirmed a thin layer of PEG on the nanocarrier surface. Shifting the zeta potential in the biological pH window from −23.9 mV (for Fe3O4) to ≈ +11 mV (for PEG/Ca(II)/Fe3O4) confirms the MNGs surface protonation. Cytotoxicity results show that cell viability and proliferation were not hindered in a wide range of nanocarrier concentrations and different incubation times.Conclusion: PEGylated calcium doped magnetic nanograin enhanced PUC19 plasmid expression into E. Coli and GFP protein expression in HEK-293 T cells compared to control. A polymerase chain reaction of the NeoR test shows that the transformed plasmids are of high quality.Keywords: gene expression, calcium doped magnetic nanograins, cytotoxicityHashemi EMahdavi HKhezri JRazi FShamsara MFarmany ADove Medical Pressarticlegene expressioncalcium doped magnetic nanograinscytotoxicityMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 14, Pp 9879-9891 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	gene expression calcium doped magnetic nanograins cytotoxicity Medicine (General) R5-920
spellingShingle	gene expression calcium doped magnetic nanograins cytotoxicity Medicine (General) R5-920 Hashemi E Mahdavi H Khezri J Razi F Shamsara M Farmany A Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin
description	Ehsan Hashemi,1,2 Hossein Mahdavi,3 Jafar Khezri,1 Farideh Razi,2,4 Mehdi Shamsara,1 Abbas Farmany5 1National Research Center for Transgenic Mouse & Animal Biotechnology Division, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran; 2Endocrinology and Metabolism Research Center, Endocrinology and Metabolism Clinical Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran; 3School of Chemistry, College of Science, University of Tehran, Tehran, Iran; 4Diabetes Research Center, Endocrinology and Metabolism Clinical Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran; 5Dental Implant Research Center, Hamadan University of Medical Sciences, Hamadan, IranCorrespondence: Abbas FarmanyDental Implant Research Center, Hamadan University of Medical Sciences, Hamadan, IranEmail a.farmany@ut.ac.irBackground: Beyond viral carriers which have been widely used in gene delivery, non-viral carriers can further improve the delivery process. However, the high cytotoxicity and low efficiency impedes the clinical application of non-viral systems. Therefore, in this work, we fabricated polyethylene glycol (PEG) coated, calcium doped magnetic nanograin (PEG/Ca(II)/Fe3O4) as a genome expression enhancer.Methods: Monodisperse magnetic nanograins (MNGs) with tunable size were synthesized by a solvothermal method. The citrate anions on the spherical surface of MNGs capture Ca2+ ions by an ion exchange process, which was followed by surface capping with PEG. The synthesized PEG/Ca(II)/Fe3O4 was characterized using Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) spectra, vibrating sample magnetometer (VSM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). MTT test was utilized to assess the toxicity of PEG/Ca(II)/Fe3O4. Real time qPCR was applied for quantification of gene expression.Results: DLS spectra and TEM images confirmed a thin layer of PEG on the nanocarrier surface. Shifting the zeta potential in the biological pH window from −23.9 mV (for Fe3O4) to ≈ +11 mV (for PEG/Ca(II)/Fe3O4) confirms the MNGs surface protonation. Cytotoxicity results show that cell viability and proliferation were not hindered in a wide range of nanocarrier concentrations and different incubation times.Conclusion: PEGylated calcium doped magnetic nanograin enhanced PUC19 plasmid expression into E. Coli and GFP protein expression in HEK-293 T cells compared to control. A polymerase chain reaction of the NeoR test shows that the transformed plasmids are of high quality.Keywords: gene expression, calcium doped magnetic nanograins, cytotoxicity
format	article
author	Hashemi E Mahdavi H Khezri J Razi F Shamsara M Farmany A
author_facet	Hashemi E Mahdavi H Khezri J Razi F Shamsara M Farmany A
author_sort	Hashemi E
title	Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin
title_short	Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin
title_full	Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin
title_fullStr	Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin
title_full_unstemmed	Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin
title_sort	enhanced gene delivery in bacterial and mammalian cells using pegylated calcium doped magnetic nanograin
publisher	Dove Medical Press
publishDate	2019
url	https://doaj.org/article/f2445cc6cecf49a3b669d44f00551eed
work_keys_str_mv	AT hashemie enhancedgenedeliveryinbacterialandmammaliancellsusingpegylatedcalciumdopedmagneticnanograin AT mahdavih enhancedgenedeliveryinbacterialandmammaliancellsusingpegylatedcalciumdopedmagneticnanograin AT khezrij enhancedgenedeliveryinbacterialandmammaliancellsusingpegylatedcalciumdopedmagneticnanograin AT razif enhancedgenedeliveryinbacterialandmammaliancellsusingpegylatedcalciumdopedmagneticnanograin AT shamsaram enhancedgenedeliveryinbacterialandmammaliancellsusingpegylatedcalciumdopedmagneticnanograin AT farmanya enhancedgenedeliveryinbacterialandmammaliancellsusingpegylatedcalciumdopedmagneticnanograin
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Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin

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