Cargando…

PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation

Over the past decade, gene therapies have attracted much attention for the development of treatments for various conditions, including cancer, neurodegenerative diseases, protein deficiencies, and autoimmune disorders. Despite the benefits of this approach, several challenges are yet to be solved to...

Descripción completa

Detalles Bibliográficos
Autores principales: Ramírez-Acosta, Carlos M., Cifuentes, Javier, Castellanos, Maria Claudia, Moreno, Rodolfo José, Muñoz-Camargo, Carolina, Cruz, Juan C., Reyes, Luis H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7356180/
https://www.ncbi.nlm.nih.gov/pubmed/32560390
http://dx.doi.org/10.3390/pharmaceutics12060561
_version_ 1783558440828796928
author Ramírez-Acosta, Carlos M.
Cifuentes, Javier
Castellanos, Maria Claudia
Moreno, Rodolfo José
Muñoz-Camargo, Carolina
Cruz, Juan C.
Reyes, Luis H.
author_facet Ramírez-Acosta, Carlos M.
Cifuentes, Javier
Castellanos, Maria Claudia
Moreno, Rodolfo José
Muñoz-Camargo, Carolina
Cruz, Juan C.
Reyes, Luis H.
author_sort Ramírez-Acosta, Carlos M.
collection PubMed
description Over the past decade, gene therapies have attracted much attention for the development of treatments for various conditions, including cancer, neurodegenerative diseases, protein deficiencies, and autoimmune disorders. Despite the benefits of this approach, several challenges are yet to be solved to reach clinical implementation. Some of these challenges include low transfection rates, limited stability under physiological conditions, and low specificity towards the target cells. An avenue to overcome such issues is to deliver the therapies with the aid of potent cell-penetrating vectors. Non-viral vectors, such as nanostructured materials, have been successfully tested in drug and gene delivery. Here, we propose the development and in vitro evaluation of a nanostructured cell-penetrating vehicle based on core/shell, magnetite/silver nanoparticles. A subsequent conjugation of a pH-responsive polymer was used to assure that the vehicle can carry and release circular DNA. Additionally, the translocating peptide Buforin II was conjugated with the aid of a polyether amine polymer to facilitate translocation and endosome escape. The obtained nanobioconjugates (magnetite/silver-pDMAEMA-PEA-BUFII) were characterized by UV-Vis spectrophotometry, dynamic light scattering (DLS), thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), scanning electron microscope equipped with energy dispersive spectroscopy (SEM+EDS), and transmission electron microscopy (TEM). They were also encapsulated in lecithin liposomes to form magnetoliposomes. The cell viability of Vero cells in the presence of the nanobioconjugates was above 95% and declined to 80% for the magnetoliposomes. The hemolytic tendency of nanobioconjugates and magnetoliposomes was below 10%, while the platelet aggregation approached that of the negative control (i.e., 35%). Cytoplasm coverage values of about 50% for both Vero and neuroblastoma cells confirmed significant cell penetration. Pearson’s correlation coefficients for both cell lines allowed us to estimate 20–40% colocalization of the nanobioconjugates with lysotracker green, which implied high levels of endosomal escape. The developed vehicles were also capable of loading around 16% of the added DNA and releasing such cargo with 8% efficiency. The developed nanoplatform holds a significant promise to enable highly efficient gene therapies as it overcomes some of the major issues associated with their eventual translation to the pre-clinical and clinical scale.
format Online
Article
Text
id pubmed-7356180
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-73561802020-07-31 PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation Ramírez-Acosta, Carlos M. Cifuentes, Javier Castellanos, Maria Claudia Moreno, Rodolfo José Muñoz-Camargo, Carolina Cruz, Juan C. Reyes, Luis H. Pharmaceutics Article Over the past decade, gene therapies have attracted much attention for the development of treatments for various conditions, including cancer, neurodegenerative diseases, protein deficiencies, and autoimmune disorders. Despite the benefits of this approach, several challenges are yet to be solved to reach clinical implementation. Some of these challenges include low transfection rates, limited stability under physiological conditions, and low specificity towards the target cells. An avenue to overcome such issues is to deliver the therapies with the aid of potent cell-penetrating vectors. Non-viral vectors, such as nanostructured materials, have been successfully tested in drug and gene delivery. Here, we propose the development and in vitro evaluation of a nanostructured cell-penetrating vehicle based on core/shell, magnetite/silver nanoparticles. A subsequent conjugation of a pH-responsive polymer was used to assure that the vehicle can carry and release circular DNA. Additionally, the translocating peptide Buforin II was conjugated with the aid of a polyether amine polymer to facilitate translocation and endosome escape. The obtained nanobioconjugates (magnetite/silver-pDMAEMA-PEA-BUFII) were characterized by UV-Vis spectrophotometry, dynamic light scattering (DLS), thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), scanning electron microscope equipped with energy dispersive spectroscopy (SEM+EDS), and transmission electron microscopy (TEM). They were also encapsulated in lecithin liposomes to form magnetoliposomes. The cell viability of Vero cells in the presence of the nanobioconjugates was above 95% and declined to 80% for the magnetoliposomes. The hemolytic tendency of nanobioconjugates and magnetoliposomes was below 10%, while the platelet aggregation approached that of the negative control (i.e., 35%). Cytoplasm coverage values of about 50% for both Vero and neuroblastoma cells confirmed significant cell penetration. Pearson’s correlation coefficients for both cell lines allowed us to estimate 20–40% colocalization of the nanobioconjugates with lysotracker green, which implied high levels of endosomal escape. The developed vehicles were also capable of loading around 16% of the added DNA and releasing such cargo with 8% efficiency. The developed nanoplatform holds a significant promise to enable highly efficient gene therapies as it overcomes some of the major issues associated with their eventual translation to the pre-clinical and clinical scale. MDPI 2020-06-17 /pmc/articles/PMC7356180/ /pubmed/32560390 http://dx.doi.org/10.3390/pharmaceutics12060561 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ramírez-Acosta, Carlos M.
Cifuentes, Javier
Castellanos, Maria Claudia
Moreno, Rodolfo José
Muñoz-Camargo, Carolina
Cruz, Juan C.
Reyes, Luis H.
PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation
title PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation
title_full PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation
title_fullStr PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation
title_full_unstemmed PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation
title_short PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation
title_sort ph-responsive, cell-penetrating, core/shell magnetite/silver nanoparticles for the delivery of plasmids: preparation, characterization, and preliminary in vitro evaluation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7356180/
https://www.ncbi.nlm.nih.gov/pubmed/32560390
http://dx.doi.org/10.3390/pharmaceutics12060561
work_keys_str_mv AT ramirezacostacarlosm phresponsivecellpenetratingcoreshellmagnetitesilvernanoparticlesforthedeliveryofplasmidspreparationcharacterizationandpreliminaryinvitroevaluation
AT cifuentesjavier phresponsivecellpenetratingcoreshellmagnetitesilvernanoparticlesforthedeliveryofplasmidspreparationcharacterizationandpreliminaryinvitroevaluation
AT castellanosmariaclaudia phresponsivecellpenetratingcoreshellmagnetitesilvernanoparticlesforthedeliveryofplasmidspreparationcharacterizationandpreliminaryinvitroevaluation
AT morenorodolfojose phresponsivecellpenetratingcoreshellmagnetitesilvernanoparticlesforthedeliveryofplasmidspreparationcharacterizationandpreliminaryinvitroevaluation
AT munozcamargocarolina phresponsivecellpenetratingcoreshellmagnetitesilvernanoparticlesforthedeliveryofplasmidspreparationcharacterizationandpreliminaryinvitroevaluation
AT cruzjuanc phresponsivecellpenetratingcoreshellmagnetitesilvernanoparticlesforthedeliveryofplasmidspreparationcharacterizationandpreliminaryinvitroevaluation
AT reyesluish phresponsivecellpenetratingcoreshellmagnetitesilvernanoparticlesforthedeliveryofplasmidspreparationcharacterizationandpreliminaryinvitroevaluation