Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model

Michelle Fraga,1,2 Talita Giacomet de Carvalho,2,3 Juliana Bidone,1 Roselena Silvestri Schuh,1,2 Ursula Matte,2,3 Helder Ferreira Teixeira1 1Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Cl&ia...

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Main Authors: Fraga M, de Carvalho TG, Bidone J, Schuh RS, Matte U, Teixeira HF
Format: Article
Language:English
Published: Dove Medical Press 2017-03-01
Series:International Journal of Nanomedicine
Subjects:
Online Access:https://www.dovepress.com/factors-influencing-transfection-efficiency-of-piduananoemulsion-compl-peer-reviewed-article-IJN
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spelling doaj-56d3c5511bd34b378d8f2e6e8732a0852020-11-25T01:06:07ZengDove Medical PressInternational Journal of Nanomedicine1178-20132017-03-01Volume 122061206731877Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine modelFraga Mde Carvalho TGBidone JSchuh RSMatte UTeixeira HFMichelle Fraga,1,2 Talita Giacomet de Carvalho,2,3 Juliana Bidone,1 Roselena Silvestri Schuh,1,2 Ursula Matte,2,3 Helder Ferreira Teixeira1 1Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre, 3Genetics and Molecular Biology Graduate Program, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil Abstract: Mucopolysaccharidosis type I (MPS I) is an autosomal disease caused by alpha-L-iduronidase (IDUA) deficiency. This study used IDUA knockout mice as a model to evaluate whether parameters such as dose of plasmid and time of treatment could influence the transfection efficiency of complexes formed with PEGylated cationic nanoemulsions and plasmid (pIDUA), which contains the gene that encodes for IDUA. Formulations were composed of medium chain triglycerides, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(amino[polyethylene glycol]-2000), 1,2-dioleoyl-sn-glycero-3-trimethylammonium propane (DOTAP), glycerol, and water and were prepared by the adsorption or encapsulation of preformed pIDUA–DOTAP complexes by high-pressure homogenization. A progressive increase in IDUA expression was observed with an increase in the dose and time of transfection for mice treated with both complexes (adsorbed and encapsulated), especially in the liver. Regardless of the complex administered, a significant increase in IDUA activity was detected in lungs and liver compared with nontreated MPS I when a dose of 60 µg was administered and IDUA activity was measured 7 days postadministration. Tissue sections of major organs showed no presence of cell necrosis, inflammatory infiltrate, or an increase in apoptosis. Furthermore, immunohistochemistry for CD68 showed no difference in the number of macrophage cells in treated and nontreated animals, indicating the absence of inflammatory reaction caused by the treatment. The data set obtained in this study allowed establishing that factors such as dose and time can influence transfection efficiency in different degrees and that these complexes did not lead to any lethal effect in the MPS I murine model used. Keywords: cationic nanoemulsions, DSPE-PEG, MPS I, plasmid, pIDUAhttps://www.dovepress.com/factors-influencing-transfection-efficiency-of-piduananoemulsion-compl-peer-reviewed-article-IJNCationic nanoemulsionsDSPE-PEGMPS IplasmidpIDUA.
collection DOAJ
language English
format Article
sources DOAJ
author Fraga M
de Carvalho TG
Bidone J
Schuh RS
Matte U
Teixeira HF
spellingShingle Fraga M
de Carvalho TG
Bidone J
Schuh RS
Matte U
Teixeira HF
Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
International Journal of Nanomedicine
Cationic nanoemulsions
DSPE-PEG
MPS I
plasmid
pIDUA.
author_facet Fraga M
de Carvalho TG
Bidone J
Schuh RS
Matte U
Teixeira HF
author_sort Fraga M
title Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_short Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_full Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_fullStr Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_full_unstemmed Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_sort factors influencing transfection efficiency of pidua/nanoemulsion complexes in a mucopolysaccharidosis type i murine model
publisher Dove Medical Press
series International Journal of Nanomedicine
issn 1178-2013
publishDate 2017-03-01
description Michelle Fraga,1,2 Talita Giacomet de Carvalho,2,3 Juliana Bidone,1 Roselena Silvestri Schuh,1,2 Ursula Matte,2,3 Helder Ferreira Teixeira1 1Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre, 3Genetics and Molecular Biology Graduate Program, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil Abstract: Mucopolysaccharidosis type I (MPS I) is an autosomal disease caused by alpha-L-iduronidase (IDUA) deficiency. This study used IDUA knockout mice as a model to evaluate whether parameters such as dose of plasmid and time of treatment could influence the transfection efficiency of complexes formed with PEGylated cationic nanoemulsions and plasmid (pIDUA), which contains the gene that encodes for IDUA. Formulations were composed of medium chain triglycerides, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(amino[polyethylene glycol]-2000), 1,2-dioleoyl-sn-glycero-3-trimethylammonium propane (DOTAP), glycerol, and water and were prepared by the adsorption or encapsulation of preformed pIDUA–DOTAP complexes by high-pressure homogenization. A progressive increase in IDUA expression was observed with an increase in the dose and time of transfection for mice treated with both complexes (adsorbed and encapsulated), especially in the liver. Regardless of the complex administered, a significant increase in IDUA activity was detected in lungs and liver compared with nontreated MPS I when a dose of 60 µg was administered and IDUA activity was measured 7 days postadministration. Tissue sections of major organs showed no presence of cell necrosis, inflammatory infiltrate, or an increase in apoptosis. Furthermore, immunohistochemistry for CD68 showed no difference in the number of macrophage cells in treated and nontreated animals, indicating the absence of inflammatory reaction caused by the treatment. The data set obtained in this study allowed establishing that factors such as dose and time can influence transfection efficiency in different degrees and that these complexes did not lead to any lethal effect in the MPS I murine model used. Keywords: cationic nanoemulsions, DSPE-PEG, MPS I, plasmid, pIDUA
topic Cationic nanoemulsions
DSPE-PEG
MPS I
plasmid
pIDUA.
url https://www.dovepress.com/factors-influencing-transfection-efficiency-of-piduananoemulsion-compl-peer-reviewed-article-IJN
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