In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles

Fahima M Hashem,1 Mohamed Nasr,1 Ahmed Khairy,2 Abdulmalik Alqurshi31Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Helwan University, Cairo, Egypt; 2Department of Pharmaceutics, National Organization for Drug Control and Research (NODCAR), Cairo, Egypt; 3Department of Pha...

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Autores principales:	Hashem FM, Nasr M, Khairy A, Alqurshi A
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2019
Materias:	p53 gene Chitosan-sodium deoxycholate nanoparticles in vitro cytotoxicity gene transfection Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/8559e034b2b94b1e88042f8b3c63c520
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spelling	oai:doaj.org-article:8559e034b2b94b1e88042f8b3c63c5202021-12-02T02:10:49ZIn vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles1178-2013https://doaj.org/article/8559e034b2b94b1e88042f8b3c63c5202019-06-01T00:00:00Zhttps://www.dovepress.com/in-vitro-cytotoxicity-and-transfection-efficiency-of-pdna-encoded-p53--peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Fahima M Hashem,1 Mohamed Nasr,1 Ahmed Khairy,2 Abdulmalik Alqurshi31Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Helwan University, Cairo, Egypt; 2Department of Pharmaceutics, National Organization for Drug Control and Research (NODCAR), Cairo, Egypt; 3Department of Pharmaceutics and Pharmaceutical Technology, College of Pharmacy, Taibah University, Medina, KSAPurpose: The objective of this work was to formulate a delivery system of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate (CS-DS) nanoparticles, and to evaluate their influence on in vitro cytotoxicity and transfection efficiency of p53 gene.Methods: The prepared pDNA-loaded CS-DS nanoparticles were evaluated for morphology, particle size, zeta potential, entrapment efficiency %, in vitro release, in vitro cytotoxicity, and transfection efficiency.Results: The mean particle size ranged from from 96.5 ± 11.31 to 405 ± 46.39 nm. All nanoparticles had good positive zeta potential values. Entrapment efficiency % ranged from 38.25 ± 3.25 to 94.89 ± 5.67. The agarose gel electrophoresis confirmed the strong binding between plasmid and CS. The in vitro pDNA release from nanoparticles exhibited an initial burst effect followed by a sustained drug release over a period of 6 days. In vitro cytotoxicity against human Caco-2 cells showed low cell cytotoxicity of plain CS-DS nanoparticles, while pDNA-loaded CS-DS nanoparticles showed a cytotoxic effect with increasing nanoparticles’ concentration. Gene transfection, analyzed by PCR and ELISA, showed a direct correlation between gene expression and concentration of pDNA. The highest expression of the gene was achieved with pDNA concentration of 9 μg/mL with 5.7 times increase compared to naked pDNA of the same concentration.Conclusion: The obtained results were very encouraging and offer an alternative approach to enhancing the transfection efficiency of genetic material-loaded chitosan-based delivery systems.Keywords: p53 gene, chitosan-sodium deoxycholate nanoparticles, in vitro cytotoxicity, gene transfectionHashem FMNasr MKhairy AAlqurshi ADove Medical Pressarticlep53 geneChitosan-sodium deoxycholate nanoparticlesin vitro cytotoxicitygene transfectionMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 14, Pp 4123-4131 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	p53 gene Chitosan-sodium deoxycholate nanoparticles in vitro cytotoxicity gene transfection Medicine (General) R5-920
spellingShingle	p53 gene Chitosan-sodium deoxycholate nanoparticles in vitro cytotoxicity gene transfection Medicine (General) R5-920 Hashem FM Nasr M Khairy A Alqurshi A In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles
description	Fahima M Hashem,1 Mohamed Nasr,1 Ahmed Khairy,2 Abdulmalik Alqurshi31Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Helwan University, Cairo, Egypt; 2Department of Pharmaceutics, National Organization for Drug Control and Research (NODCAR), Cairo, Egypt; 3Department of Pharmaceutics and Pharmaceutical Technology, College of Pharmacy, Taibah University, Medina, KSAPurpose: The objective of this work was to formulate a delivery system of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate (CS-DS) nanoparticles, and to evaluate their influence on in vitro cytotoxicity and transfection efficiency of p53 gene.Methods: The prepared pDNA-loaded CS-DS nanoparticles were evaluated for morphology, particle size, zeta potential, entrapment efficiency %, in vitro release, in vitro cytotoxicity, and transfection efficiency.Results: The mean particle size ranged from from 96.5 ± 11.31 to 405 ± 46.39 nm. All nanoparticles had good positive zeta potential values. Entrapment efficiency % ranged from 38.25 ± 3.25 to 94.89 ± 5.67. The agarose gel electrophoresis confirmed the strong binding between plasmid and CS. The in vitro pDNA release from nanoparticles exhibited an initial burst effect followed by a sustained drug release over a period of 6 days. In vitro cytotoxicity against human Caco-2 cells showed low cell cytotoxicity of plain CS-DS nanoparticles, while pDNA-loaded CS-DS nanoparticles showed a cytotoxic effect with increasing nanoparticles’ concentration. Gene transfection, analyzed by PCR and ELISA, showed a direct correlation between gene expression and concentration of pDNA. The highest expression of the gene was achieved with pDNA concentration of 9 μg/mL with 5.7 times increase compared to naked pDNA of the same concentration.Conclusion: The obtained results were very encouraging and offer an alternative approach to enhancing the transfection efficiency of genetic material-loaded chitosan-based delivery systems.Keywords: p53 gene, chitosan-sodium deoxycholate nanoparticles, in vitro cytotoxicity, gene transfection
format	article
author	Hashem FM Nasr M Khairy A Alqurshi A
author_facet	Hashem FM Nasr M Khairy A Alqurshi A
author_sort	Hashem FM
title	In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles
title_short	In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles
title_full	In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles
title_fullStr	In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles
title_full_unstemmed	In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles
title_sort	in vitro cytotoxicity and transfection efficiency of pdna encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles
publisher	Dove Medical Press
publishDate	2019
url	https://doaj.org/article/8559e034b2b94b1e88042f8b3c63c520
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In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles

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