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Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery
In recent years, lipid nanoparticles (LNPs) have gained considerable attention in numerous research fields ranging from gene therapy to cancer immunotherapy and DNA vaccination. While some RNA-encapsulating LNP formulations passed clinical trials, DNA-loaded LNPs have been only marginally explored s...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8400491/ https://www.ncbi.nlm.nih.gov/pubmed/34452253 http://dx.doi.org/10.3390/pharmaceutics13081292 |
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author | Quagliarini, Erica Renzi, Serena Digiacomo, Luca Giulimondi, Francesca Sartori, Barbara Amenitsch, Heinz Tassinari, Valentina Masuelli, Laura Bei, Roberto Cui, Lishan Wang, Junbiao Amici, Augusto Marchini, Cristina Pozzi, Daniela Caracciolo, Giulio |
author_facet | Quagliarini, Erica Renzi, Serena Digiacomo, Luca Giulimondi, Francesca Sartori, Barbara Amenitsch, Heinz Tassinari, Valentina Masuelli, Laura Bei, Roberto Cui, Lishan Wang, Junbiao Amici, Augusto Marchini, Cristina Pozzi, Daniela Caracciolo, Giulio |
author_sort | Quagliarini, Erica |
collection | PubMed |
description | In recent years, lipid nanoparticles (LNPs) have gained considerable attention in numerous research fields ranging from gene therapy to cancer immunotherapy and DNA vaccination. While some RNA-encapsulating LNP formulations passed clinical trials, DNA-loaded LNPs have been only marginally explored so far. To fulfil this gap, herein we investigated the effect of several factors influencing the microfluidic formulation and transfection behavior of DNA-loaded LNPs such as PEGylation, total flow rate (TFR), concentration and particle density at the cell surface. We show that PEGylation and post-synthesis sample concentration facilitated formulation of homogeneous and small size LNPs with high transfection efficiency and minor, if any, cytotoxicity on human Embryonic Kidney293 (HEK-293), spontaneously immortalized human keratinocytes (HaCaT), immortalized keratinocytes (N/TERT) generated from the transduction of human primary keratinocytes, and epidermoid cervical cancer (CaSki) cell lines. On the other side, increasing TFR had a detrimental effect both on the physicochemical properties and transfection properties of LNPs. Lastly, the effect of particle concentration at the cell surface on the transfection efficiency (TE) and cell viability was largely dependent on the cell line, suggesting that its case-by-case optimization would be necessary. Overall, we demonstrate that fine tuning formulation and microfluidic parameters is a vital step for the generation of highly efficient DNA-loaded LNPs. |
format | Online Article Text |
id | pubmed-8400491 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-84004912021-08-29 Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery Quagliarini, Erica Renzi, Serena Digiacomo, Luca Giulimondi, Francesca Sartori, Barbara Amenitsch, Heinz Tassinari, Valentina Masuelli, Laura Bei, Roberto Cui, Lishan Wang, Junbiao Amici, Augusto Marchini, Cristina Pozzi, Daniela Caracciolo, Giulio Pharmaceutics Article In recent years, lipid nanoparticles (LNPs) have gained considerable attention in numerous research fields ranging from gene therapy to cancer immunotherapy and DNA vaccination. While some RNA-encapsulating LNP formulations passed clinical trials, DNA-loaded LNPs have been only marginally explored so far. To fulfil this gap, herein we investigated the effect of several factors influencing the microfluidic formulation and transfection behavior of DNA-loaded LNPs such as PEGylation, total flow rate (TFR), concentration and particle density at the cell surface. We show that PEGylation and post-synthesis sample concentration facilitated formulation of homogeneous and small size LNPs with high transfection efficiency and minor, if any, cytotoxicity on human Embryonic Kidney293 (HEK-293), spontaneously immortalized human keratinocytes (HaCaT), immortalized keratinocytes (N/TERT) generated from the transduction of human primary keratinocytes, and epidermoid cervical cancer (CaSki) cell lines. On the other side, increasing TFR had a detrimental effect both on the physicochemical properties and transfection properties of LNPs. Lastly, the effect of particle concentration at the cell surface on the transfection efficiency (TE) and cell viability was largely dependent on the cell line, suggesting that its case-by-case optimization would be necessary. Overall, we demonstrate that fine tuning formulation and microfluidic parameters is a vital step for the generation of highly efficient DNA-loaded LNPs. MDPI 2021-08-19 /pmc/articles/PMC8400491/ /pubmed/34452253 http://dx.doi.org/10.3390/pharmaceutics13081292 Text en © 2021 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 Quagliarini, Erica Renzi, Serena Digiacomo, Luca Giulimondi, Francesca Sartori, Barbara Amenitsch, Heinz Tassinari, Valentina Masuelli, Laura Bei, Roberto Cui, Lishan Wang, Junbiao Amici, Augusto Marchini, Cristina Pozzi, Daniela Caracciolo, Giulio Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery |
title | Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery |
title_full | Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery |
title_fullStr | Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery |
title_full_unstemmed | Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery |
title_short | Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery |
title_sort | microfluidic formulation of dna-loaded multicomponent lipid nanoparticles for gene delivery |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8400491/ https://www.ncbi.nlm.nih.gov/pubmed/34452253 http://dx.doi.org/10.3390/pharmaceutics13081292 |
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