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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...

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Autores principales: 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
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
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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.
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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
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