Cargando…
Choosing an Optimal Solvent Is Crucial for Obtaining Cell-Penetrating Peptide Nanoparticles with Desired Properties and High Activity in Nucleic Acid Delivery
Cell-penetrating peptides (CPPs) are highly promising transfection agents that can deliver various compounds into living cells, including nucleic acids (NAs). Positively charged CPPs can form non-covalent complexes with negatively charged NAs, enabling simple and time-efficient nanoparticle preparat...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9963036/ https://www.ncbi.nlm.nih.gov/pubmed/36839718 http://dx.doi.org/10.3390/pharmaceutics15020396 |
_version_ | 1784896151400480768 |
---|---|
author | Biswas, Abhijit Maloverjan, Maria Padari, Kärt Abroi, Aare Rätsep, Margus Wärmländer, Sebastian K. T. S. Jarvet, Jüri Gräslund, Astrid Kisand, Vambola Lõhmus, Rünno Pooga, Margus |
author_facet | Biswas, Abhijit Maloverjan, Maria Padari, Kärt Abroi, Aare Rätsep, Margus Wärmländer, Sebastian K. T. S. Jarvet, Jüri Gräslund, Astrid Kisand, Vambola Lõhmus, Rünno Pooga, Margus |
author_sort | Biswas, Abhijit |
collection | PubMed |
description | Cell-penetrating peptides (CPPs) are highly promising transfection agents that can deliver various compounds into living cells, including nucleic acids (NAs). Positively charged CPPs can form non-covalent complexes with negatively charged NAs, enabling simple and time-efficient nanoparticle preparation. However, as CPPs have substantially different chemical and physical properties, their complexation with the cargo and characteristics of the resulting nanoparticles largely depends on the properties of the surrounding environment, i.e., solution. Here, we show that the solvent used for the initial dissolving of a CPP determines the properties of the resulting CPP particles formed in an aqueous solution, including the activity and toxicity of the CPP–NA complexes. Using different biophysical methods such as dynamic light scattering (DLS), atomic force microscopy (AFM), transmission and scanning electron microscopy (TEM and SEM), we show that PepFect14 (PF14), a cationic amphipathic CPP, forms spherical particles of uniform size when dissolved in organic solvents, such as ethanol and DMSO. Water-dissolved PF14, however, tends to form micelles and non-uniform aggregates. When dissolved in organic solvents, PF14 retains its α-helical conformation and biological activity in cell culture conditions without any increase in cytotoxicity. Altogether, our results indicate that by using a solvent that matches the chemical nature of the CPP, the properties of the peptide–cargo particles can be tuned in the desired way. This can be of critical importance for in vivo applications, where CPP particles that are too large, non-uniform, or prone to aggregation may induce severe consequences. |
format | Online Article Text |
id | pubmed-9963036 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-99630362023-02-26 Choosing an Optimal Solvent Is Crucial for Obtaining Cell-Penetrating Peptide Nanoparticles with Desired Properties and High Activity in Nucleic Acid Delivery Biswas, Abhijit Maloverjan, Maria Padari, Kärt Abroi, Aare Rätsep, Margus Wärmländer, Sebastian K. T. S. Jarvet, Jüri Gräslund, Astrid Kisand, Vambola Lõhmus, Rünno Pooga, Margus Pharmaceutics Article Cell-penetrating peptides (CPPs) are highly promising transfection agents that can deliver various compounds into living cells, including nucleic acids (NAs). Positively charged CPPs can form non-covalent complexes with negatively charged NAs, enabling simple and time-efficient nanoparticle preparation. However, as CPPs have substantially different chemical and physical properties, their complexation with the cargo and characteristics of the resulting nanoparticles largely depends on the properties of the surrounding environment, i.e., solution. Here, we show that the solvent used for the initial dissolving of a CPP determines the properties of the resulting CPP particles formed in an aqueous solution, including the activity and toxicity of the CPP–NA complexes. Using different biophysical methods such as dynamic light scattering (DLS), atomic force microscopy (AFM), transmission and scanning electron microscopy (TEM and SEM), we show that PepFect14 (PF14), a cationic amphipathic CPP, forms spherical particles of uniform size when dissolved in organic solvents, such as ethanol and DMSO. Water-dissolved PF14, however, tends to form micelles and non-uniform aggregates. When dissolved in organic solvents, PF14 retains its α-helical conformation and biological activity in cell culture conditions without any increase in cytotoxicity. Altogether, our results indicate that by using a solvent that matches the chemical nature of the CPP, the properties of the peptide–cargo particles can be tuned in the desired way. This can be of critical importance for in vivo applications, where CPP particles that are too large, non-uniform, or prone to aggregation may induce severe consequences. MDPI 2023-01-24 /pmc/articles/PMC9963036/ /pubmed/36839718 http://dx.doi.org/10.3390/pharmaceutics15020396 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Biswas, Abhijit Maloverjan, Maria Padari, Kärt Abroi, Aare Rätsep, Margus Wärmländer, Sebastian K. T. S. Jarvet, Jüri Gräslund, Astrid Kisand, Vambola Lõhmus, Rünno Pooga, Margus Choosing an Optimal Solvent Is Crucial for Obtaining Cell-Penetrating Peptide Nanoparticles with Desired Properties and High Activity in Nucleic Acid Delivery |
title | Choosing an Optimal Solvent Is Crucial for Obtaining Cell-Penetrating Peptide Nanoparticles with Desired Properties and High Activity in Nucleic Acid Delivery |
title_full | Choosing an Optimal Solvent Is Crucial for Obtaining Cell-Penetrating Peptide Nanoparticles with Desired Properties and High Activity in Nucleic Acid Delivery |
title_fullStr | Choosing an Optimal Solvent Is Crucial for Obtaining Cell-Penetrating Peptide Nanoparticles with Desired Properties and High Activity in Nucleic Acid Delivery |
title_full_unstemmed | Choosing an Optimal Solvent Is Crucial for Obtaining Cell-Penetrating Peptide Nanoparticles with Desired Properties and High Activity in Nucleic Acid Delivery |
title_short | Choosing an Optimal Solvent Is Crucial for Obtaining Cell-Penetrating Peptide Nanoparticles with Desired Properties and High Activity in Nucleic Acid Delivery |
title_sort | choosing an optimal solvent is crucial for obtaining cell-penetrating peptide nanoparticles with desired properties and high activity in nucleic acid delivery |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9963036/ https://www.ncbi.nlm.nih.gov/pubmed/36839718 http://dx.doi.org/10.3390/pharmaceutics15020396 |
work_keys_str_mv | AT biswasabhijit choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT maloverjanmaria choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT padarikart choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT abroiaare choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT ratsepmargus choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT warmlandersebastiankts choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT jarvetjuri choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT graslundastrid choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT kisandvambola choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT lohmusrunno choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery AT poogamargus choosinganoptimalsolventiscrucialforobtainingcellpenetratingpeptidenanoparticleswithdesiredpropertiesandhighactivityinnucleicaciddelivery |