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Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres
One of the first methods to encapsulate drugs within polymer nanospheres was developed by Fessi and coworkers in 1989 and consisted of one-step nanoprecipitation based on solvent displacement. However, proteins are poorly encapsulated within polymer nanoparticles using this method because of their l...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3541529/ https://www.ncbi.nlm.nih.gov/pubmed/23316451 http://dx.doi.org/10.1016/j.rinphs.2012.11.001 |
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author | Morales-Cruz, Moraima Flores-Fernández, Giselle M. Morales-Cruz, Myreisa Orellano, Elsie A. Rodriguez-Martinez, José A. Ruiz, Mercedes Griebenow, Kai |
author_facet | Morales-Cruz, Moraima Flores-Fernández, Giselle M. Morales-Cruz, Myreisa Orellano, Elsie A. Rodriguez-Martinez, José A. Ruiz, Mercedes Griebenow, Kai |
author_sort | Morales-Cruz, Moraima |
collection | PubMed |
description | One of the first methods to encapsulate drugs within polymer nanospheres was developed by Fessi and coworkers in 1989 and consisted of one-step nanoprecipitation based on solvent displacement. However, proteins are poorly encapsulated within polymer nanoparticles using this method because of their limited solubility in organic solvents. To overcome this limitation, we developed a two-step nanoprecipitation method and encapsulated various proteins with high efficiency into poly(lactic-co-glycolic)acid (PLGA) nanospheres (NP). In this method, a protein nanoprecipitation step is used first followed by a second polymer nanoprecipitation step. Two model enzymes, lysozyme and α-chymotrypsin, were used for the optimization of the method. We obtained encapsulation efficiencies of >70%, an amount of buffer-insoluble protein aggregates of typically <2%, and a high residual activity of typically >90%. The optimum conditions identified for lysozyme were used to successfully encapsulate cytochrome c(Cyt-c), an apoptosis-initiating basic protein of similar size, to verify reproducibility of the encapsulation procedure. The size of the Cyt-c loaded-PLGA nanospheres was around 300–400 nm indicating the potential of the delivery system to passively target tumors. Cell viability studies, using a human cervical cancer cell line (HeLa), demonstrate excellent biocompatibility of the PLGA nanoparticles. PLGA nanoparticles carrying encapsulated Cyt-c were not efficient in causing apoptosis presumably because PLGA nanoparticles are not efficiently taken up by the cells. Future systems will have to be optimized to ascertain efficient cellular uptake of the nanoparticles by, e.g., surface modification with receptor ligands. |
format | Online Article Text |
id | pubmed-3541529 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-35415292013-01-10 Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres Morales-Cruz, Moraima Flores-Fernández, Giselle M. Morales-Cruz, Myreisa Orellano, Elsie A. Rodriguez-Martinez, José A. Ruiz, Mercedes Griebenow, Kai Results Pharma Sci Article One of the first methods to encapsulate drugs within polymer nanospheres was developed by Fessi and coworkers in 1989 and consisted of one-step nanoprecipitation based on solvent displacement. However, proteins are poorly encapsulated within polymer nanoparticles using this method because of their limited solubility in organic solvents. To overcome this limitation, we developed a two-step nanoprecipitation method and encapsulated various proteins with high efficiency into poly(lactic-co-glycolic)acid (PLGA) nanospheres (NP). In this method, a protein nanoprecipitation step is used first followed by a second polymer nanoprecipitation step. Two model enzymes, lysozyme and α-chymotrypsin, were used for the optimization of the method. We obtained encapsulation efficiencies of >70%, an amount of buffer-insoluble protein aggregates of typically <2%, and a high residual activity of typically >90%. The optimum conditions identified for lysozyme were used to successfully encapsulate cytochrome c(Cyt-c), an apoptosis-initiating basic protein of similar size, to verify reproducibility of the encapsulation procedure. The size of the Cyt-c loaded-PLGA nanospheres was around 300–400 nm indicating the potential of the delivery system to passively target tumors. Cell viability studies, using a human cervical cancer cell line (HeLa), demonstrate excellent biocompatibility of the PLGA nanoparticles. PLGA nanoparticles carrying encapsulated Cyt-c were not efficient in causing apoptosis presumably because PLGA nanoparticles are not efficiently taken up by the cells. Future systems will have to be optimized to ascertain efficient cellular uptake of the nanoparticles by, e.g., surface modification with receptor ligands. Elsevier 2012-11-24 /pmc/articles/PMC3541529/ /pubmed/23316451 http://dx.doi.org/10.1016/j.rinphs.2012.11.001 Text en © 2012 Elsevier B.V. All rights reserved. http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). |
spellingShingle | Article Morales-Cruz, Moraima Flores-Fernández, Giselle M. Morales-Cruz, Myreisa Orellano, Elsie A. Rodriguez-Martinez, José A. Ruiz, Mercedes Griebenow, Kai Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres |
title | Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres |
title_full | Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres |
title_fullStr | Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres |
title_full_unstemmed | Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres |
title_short | Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres |
title_sort | two-step nanoprecipitation for the production of protein-loaded plga nanospheres |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3541529/ https://www.ncbi.nlm.nih.gov/pubmed/23316451 http://dx.doi.org/10.1016/j.rinphs.2012.11.001 |
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