Evidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications

Elsa Patricia Rondon,1 Houda Abir Benabdoun,1 Francis Vallières,1 Maicon Segalla Petrônio,2 Marcio José Tiera,2 Mohamed Benderdour,1 Julio Cesar Fernandes1 1Orthopedic Research Laboratory, Hôpital Du Sacré-C&oelig;ur De Montréal, Un...

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Autores principales:	Rondon EP, Benabdoun HA, Vallières F, Segalla Petrônio M, Tiera MJ, Benderdour M, Fernandes JC
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2020
Materias:	chitosan nanoparticles sirna biocompatibility assays gene therapy toxicity. Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/e9b69568a5d14c55956474ae15e42d0e
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spelling	oai:doaj.org-article:e9b69568a5d14c55956474ae15e42d0e2021-12-02T08:28:17ZEvidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications1178-2013https://doaj.org/article/e9b69568a5d14c55956474ae15e42d0e2020-08-01T00:00:00Zhttps://www.dovepress.com/evidence-supporting-the-safety-of-pegylated-diethylaminoethyl-chitosan-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Elsa Patricia Rondon,1 Houda Abir Benabdoun,1 Francis Vallières,1 Maicon Segalla Petrônio,2 Marcio José Tiera,2 Mohamed Benderdour,1 Julio Cesar Fernandes1 1Orthopedic Research Laboratory, Hôpital Du Sacré-C&oelig;ur De Montréal, Université De Montréal, Montréal, Québec, Canada; 2Institute of Biosciences, Humanities and Exact Sciences, Department of Chemistry and Environmental Sciences, UNESP-São Paulo State University, São José Do Rio Preto, São Paulo State, BrazilCorrespondence: Julio Cesar Fernandes Tel +1 514-338-2222 Ext. 2489Email julio.c.fernandes@umontreal.caPurpose: Diethylaminoethyl-chitosan (DEAE-CH) is a derivative with excellent potential as a delivery vector for gene therapy applications. The aim of this study is to evaluate its toxicological profile for potential future clinical applications.Methods: An endotoxin-free chitosan (CH) modified with DEAE, folic acid (FA) and polyethylene glycol (PEG) was used to complex small interfering RNA (siRNA) and form nanoparticles (DEAE12-CH-PEG-FA2/siRNA). Based on the guidelines from the International Organization for Standardization (ISO), the American Society for Testing and Materials (ASTM), and the Nanotechnology Characterization Laboratory (NCL), we evaluated the effects of the interaction between these nanoparticles and blood components. In vitro screening assays such as hemolysis, hemagglutination, complement activation, platelet aggregation, coagulation times, cytokine production, and reactive species, such as nitric oxide (NO) and reactive oxygen species (ROS), were performed on erythrocytes, plasma, platelets, peripheral blood mononuclear cells (PBMC) and Raw 264.7 macrophages. Moreover, MTS and LDH assays on Raw 264.7 macrophages, PBMC and MG-63 cells were performed.Results: Our results show that a targeted theoretical plasma concentration (TPC) of DEAE12-CH-PEG-FA2/siRNA nanoparticles falls within the guidelines’ thresholds: < 1% hemolysis, 2.9% platelet aggregation, no complement activation, and no effect on coagulation times. ROS and NO production levels were comparable to controls. Cytokine secretion (TNF-α, IL-6, IL-4, and IL-10) was not affected by nanoparticles except for IL-1β and IL-8. Nanoparticles showed a slight agglutination. Cell viability was > 70% for TPC in all cell types, although LDH levels were statistically significant in Raw 264.7 macrophages and PBMC after 24 and 48 h of incubation.Conclusion: These DEAE12-CH-PEG-FA2/siRNA nanoparticles fulfill the existing ISO, ASTM and NCL guidelines’ threshold criteria, and their low toxicity and blood biocompatibility warrant further investigation for potential clinical applications.Keywords: chitosan, nanoparticles, siRNA, biocompatibility assays, gene therapy, toxicityRondon EPBenabdoun HAVallières FSegalla Petrônio MTiera MJBenderdour MFernandes JCDove Medical Pressarticlechitosannanoparticlessirnabiocompatibility assaysgene therapytoxicity.Medicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 15, Pp 6183-6200 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	chitosan nanoparticles sirna biocompatibility assays gene therapy toxicity. Medicine (General) R5-920
spellingShingle	chitosan nanoparticles sirna biocompatibility assays gene therapy toxicity. Medicine (General) R5-920 Rondon EP Benabdoun HA Vallières F Segalla Petrônio M Tiera MJ Benderdour M Fernandes JC Evidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications
description	Elsa Patricia Rondon,1 Houda Abir Benabdoun,1 Francis Vallières,1 Maicon Segalla Petrônio,2 Marcio José Tiera,2 Mohamed Benderdour,1 Julio Cesar Fernandes1 1Orthopedic Research Laboratory, Hôpital Du Sacré-C&oelig;ur De Montréal, Université De Montréal, Montréal, Québec, Canada; 2Institute of Biosciences, Humanities and Exact Sciences, Department of Chemistry and Environmental Sciences, UNESP-São Paulo State University, São José Do Rio Preto, São Paulo State, BrazilCorrespondence: Julio Cesar Fernandes Tel +1 514-338-2222 Ext. 2489Email julio.c.fernandes@umontreal.caPurpose: Diethylaminoethyl-chitosan (DEAE-CH) is a derivative with excellent potential as a delivery vector for gene therapy applications. The aim of this study is to evaluate its toxicological profile for potential future clinical applications.Methods: An endotoxin-free chitosan (CH) modified with DEAE, folic acid (FA) and polyethylene glycol (PEG) was used to complex small interfering RNA (siRNA) and form nanoparticles (DEAE12-CH-PEG-FA2/siRNA). Based on the guidelines from the International Organization for Standardization (ISO), the American Society for Testing and Materials (ASTM), and the Nanotechnology Characterization Laboratory (NCL), we evaluated the effects of the interaction between these nanoparticles and blood components. In vitro screening assays such as hemolysis, hemagglutination, complement activation, platelet aggregation, coagulation times, cytokine production, and reactive species, such as nitric oxide (NO) and reactive oxygen species (ROS), were performed on erythrocytes, plasma, platelets, peripheral blood mononuclear cells (PBMC) and Raw 264.7 macrophages. Moreover, MTS and LDH assays on Raw 264.7 macrophages, PBMC and MG-63 cells were performed.Results: Our results show that a targeted theoretical plasma concentration (TPC) of DEAE12-CH-PEG-FA2/siRNA nanoparticles falls within the guidelines’ thresholds: < 1% hemolysis, 2.9% platelet aggregation, no complement activation, and no effect on coagulation times. ROS and NO production levels were comparable to controls. Cytokine secretion (TNF-α, IL-6, IL-4, and IL-10) was not affected by nanoparticles except for IL-1β and IL-8. Nanoparticles showed a slight agglutination. Cell viability was > 70% for TPC in all cell types, although LDH levels were statistically significant in Raw 264.7 macrophages and PBMC after 24 and 48 h of incubation.Conclusion: These DEAE12-CH-PEG-FA2/siRNA nanoparticles fulfill the existing ISO, ASTM and NCL guidelines’ threshold criteria, and their low toxicity and blood biocompatibility warrant further investigation for potential clinical applications.Keywords: chitosan, nanoparticles, siRNA, biocompatibility assays, gene therapy, toxicity
format	article
author	Rondon EP Benabdoun HA Vallières F Segalla Petrônio M Tiera MJ Benderdour M Fernandes JC
author_facet	Rondon EP Benabdoun HA Vallières F Segalla Petrônio M Tiera MJ Benderdour M Fernandes JC
author_sort	Rondon EP
title	Evidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications
title_short	Evidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications
title_full	Evidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications
title_fullStr	Evidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications
title_full_unstemmed	Evidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications
title_sort	evidence supporting the safety of pegylated diethylaminoethyl-chitosan polymer as a nanovector for gene therapy applications
publisher	Dove Medical Press
publishDate	2020
url	https://doaj.org/article/e9b69568a5d14c55956474ae15e42d0e
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Evidence Supporting the Safety of Pegylated Diethylaminoethyl-Chitosan Polymer as a Nanovector for Gene Therapy Applications

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