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...
Autores principales: | , , , , , , |
---|---|
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 |