Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines

Juan P. Peñaloza, Valeria Márquez-Miranda, Mauricio Cabaña-Brunod, Rodrigo Reyes-Ramírez, Felipe M. Llancalahuen, Cristian Vilos, Fernanda Maldonado-Biermann, Luis A. Velásquez, Juan A. Fuentes, Fernando D. González-Nilo, Maité Rodríguez-Díaz, Carolina Otero

Resultado de la investigación: Article

  • 11 Citas

Resumen

Background: Nanotechnology is a science that involves imaging, measurement, modeling and a manipulation of matter at the nanometric scale. One application of this technology is drug delivery systems based on nanoparticles obtained from natural or synthetic sources. An example of these systems is synthetized from poly(3-hydroxybutyrate-co-3-hydroxyvalerate), which is a biodegradable, biocompatible and a low production cost polymer. The aim of this work was to investigate the uptake mechanism of PHBV nanoparticles in two different epithelial cell lines (HeLa and SKOV-3). Results: As a first step, we characterized size, shape and surface charge of nanoparticles using dynamic light scattering and transmission electron microscopy. Intracellular incorporation was evaluated through flow cytometry and fluorescence microscopy using intracellular markers. We concluded that cellular uptake mechanism is carried out in a time, concentration and energy dependent way. Our results showed that nanoparticle uptake displays a cell-specific pattern, since we have observed different colocalization in two different cell lines. In HeLa (Cervical cancer cells) this process may occur via classical endocytosis pathway and some internalization via caveolin-dependent was also observed, whereas in SKOV-3 (Ovarian cancer cells) these patterns were not observed. Rearrangement of actin filaments showed differential nanoparticle internalization patterns for HeLa and SKOV-3. Additionally, final fate of nanoparticles was also determined, showing that in both cell lines, nanoparticles ended up in lysosomes but at different times, where they are finally degraded, thereby releasing their contents. Conclusions: Our results, provide novel insight about PHBV nanoparticles internalization suggesting that for develop a proper drug delivery system is critical understand the uptake mechanism.

IdiomaEnglish
Número de artículo1
PublicaciónJournal of Nanobiotechnology
Volumen15
Número de edición1
DOI
EstadoPublished - 3 ene 2017

Huella dactilar

Nanoparticles
Epithelial Cells
Cell Line
Cells
Drug Delivery Systems
Caveolins
Nanotechnology
Flow cytometry
Fluorescence microscopy
Dynamic light scattering
Surface charge
Light transmission
Endocytosis
Lysosomes
Transmission Electron Microscopy
Actin Cytoskeleton
Fluorescence Microscopy
Uterine Cervical Neoplasms
Ovarian Neoplasms
Actins

ASJC Scopus subject areas

  • Bioengineering
  • Medicine (miscellaneous)
  • Molecular Medicine
  • Biomedical Engineering
  • Applied Microbiology and Biotechnology
  • Pharmaceutical Science

Citar esto

Peñaloza, Juan P. ; Márquez-Miranda, Valeria ; Cabaña-Brunod, Mauricio ; Reyes-Ramírez, Rodrigo ; Llancalahuen, Felipe M. ; Vilos, Cristian ; Maldonado-Biermann, Fernanda ; Velásquez, Luis A. ; Fuentes, Juan A. ; González-Nilo, Fernando D. ; Rodríguez-Díaz, Maité ; Otero, Carolina. / Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines. En: Journal of Nanobiotechnology. 2017 ; Vol. 15, N.º 1.
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abstract = "Background: Nanotechnology is a science that involves imaging, measurement, modeling and a manipulation of matter at the nanometric scale. One application of this technology is drug delivery systems based on nanoparticles obtained from natural or synthetic sources. An example of these systems is synthetized from poly(3-hydroxybutyrate-co-3-hydroxyvalerate), which is a biodegradable, biocompatible and a low production cost polymer. The aim of this work was to investigate the uptake mechanism of PHBV nanoparticles in two different epithelial cell lines (HeLa and SKOV-3). Results: As a first step, we characterized size, shape and surface charge of nanoparticles using dynamic light scattering and transmission electron microscopy. Intracellular incorporation was evaluated through flow cytometry and fluorescence microscopy using intracellular markers. We concluded that cellular uptake mechanism is carried out in a time, concentration and energy dependent way. Our results showed that nanoparticle uptake displays a cell-specific pattern, since we have observed different colocalization in two different cell lines. In HeLa (Cervical cancer cells) this process may occur via classical endocytosis pathway and some internalization via caveolin-dependent was also observed, whereas in SKOV-3 (Ovarian cancer cells) these patterns were not observed. Rearrangement of actin filaments showed differential nanoparticle internalization patterns for HeLa and SKOV-3. Additionally, final fate of nanoparticles was also determined, showing that in both cell lines, nanoparticles ended up in lysosomes but at different times, where they are finally degraded, thereby releasing their contents. Conclusions: Our results, provide novel insight about PHBV nanoparticles internalization suggesting that for develop a proper drug delivery system is critical understand the uptake mechanism.",
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Peñaloza, JP, Márquez-Miranda, V, Cabaña-Brunod, M, Reyes-Ramírez, R, Llancalahuen, FM, Vilos, C, Maldonado-Biermann, F, Velásquez, LA, Fuentes, JA, González-Nilo, FD, Rodríguez-Díaz, M & Otero, C 2017, 'Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines' Journal of Nanobiotechnology, vol. 15, n.º 1, 1. DOI: 10.1186/s12951-016-0241-6

Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines. / Peñaloza, Juan P.; Márquez-Miranda, Valeria; Cabaña-Brunod, Mauricio; Reyes-Ramírez, Rodrigo; Llancalahuen, Felipe M.; Vilos, Cristian; Maldonado-Biermann, Fernanda; Velásquez, Luis A.; Fuentes, Juan A.; González-Nilo, Fernando D.; Rodríguez-Díaz, Maité; Otero, Carolina.

En: Journal of Nanobiotechnology, Vol. 15, N.º 1, 1, 03.01.2017.

Resultado de la investigación: Article

TY - JOUR

T1 - Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines

AU - Peñaloza,Juan P.

AU - Márquez-Miranda,Valeria

AU - Cabaña-Brunod,Mauricio

AU - Reyes-Ramírez,Rodrigo

AU - Llancalahuen,Felipe M.

AU - Vilos,Cristian

AU - Maldonado-Biermann,Fernanda

AU - Velásquez,Luis A.

AU - Fuentes,Juan A.

AU - González-Nilo,Fernando D.

AU - Rodríguez-Díaz,Maité

AU - Otero,Carolina

PY - 2017/1/3

Y1 - 2017/1/3

N2 - Background: Nanotechnology is a science that involves imaging, measurement, modeling and a manipulation of matter at the nanometric scale. One application of this technology is drug delivery systems based on nanoparticles obtained from natural or synthetic sources. An example of these systems is synthetized from poly(3-hydroxybutyrate-co-3-hydroxyvalerate), which is a biodegradable, biocompatible and a low production cost polymer. The aim of this work was to investigate the uptake mechanism of PHBV nanoparticles in two different epithelial cell lines (HeLa and SKOV-3). Results: As a first step, we characterized size, shape and surface charge of nanoparticles using dynamic light scattering and transmission electron microscopy. Intracellular incorporation was evaluated through flow cytometry and fluorescence microscopy using intracellular markers. We concluded that cellular uptake mechanism is carried out in a time, concentration and energy dependent way. Our results showed that nanoparticle uptake displays a cell-specific pattern, since we have observed different colocalization in two different cell lines. In HeLa (Cervical cancer cells) this process may occur via classical endocytosis pathway and some internalization via caveolin-dependent was also observed, whereas in SKOV-3 (Ovarian cancer cells) these patterns were not observed. Rearrangement of actin filaments showed differential nanoparticle internalization patterns for HeLa and SKOV-3. Additionally, final fate of nanoparticles was also determined, showing that in both cell lines, nanoparticles ended up in lysosomes but at different times, where they are finally degraded, thereby releasing their contents. Conclusions: Our results, provide novel insight about PHBV nanoparticles internalization suggesting that for develop a proper drug delivery system is critical understand the uptake mechanism.

AB - Background: Nanotechnology is a science that involves imaging, measurement, modeling and a manipulation of matter at the nanometric scale. One application of this technology is drug delivery systems based on nanoparticles obtained from natural or synthetic sources. An example of these systems is synthetized from poly(3-hydroxybutyrate-co-3-hydroxyvalerate), which is a biodegradable, biocompatible and a low production cost polymer. The aim of this work was to investigate the uptake mechanism of PHBV nanoparticles in two different epithelial cell lines (HeLa and SKOV-3). Results: As a first step, we characterized size, shape and surface charge of nanoparticles using dynamic light scattering and transmission electron microscopy. Intracellular incorporation was evaluated through flow cytometry and fluorescence microscopy using intracellular markers. We concluded that cellular uptake mechanism is carried out in a time, concentration and energy dependent way. Our results showed that nanoparticle uptake displays a cell-specific pattern, since we have observed different colocalization in two different cell lines. In HeLa (Cervical cancer cells) this process may occur via classical endocytosis pathway and some internalization via caveolin-dependent was also observed, whereas in SKOV-3 (Ovarian cancer cells) these patterns were not observed. Rearrangement of actin filaments showed differential nanoparticle internalization patterns for HeLa and SKOV-3. Additionally, final fate of nanoparticles was also determined, showing that in both cell lines, nanoparticles ended up in lysosomes but at different times, where they are finally degraded, thereby releasing their contents. Conclusions: Our results, provide novel insight about PHBV nanoparticles internalization suggesting that for develop a proper drug delivery system is critical understand the uptake mechanism.

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Peñaloza JP, Márquez-Miranda V, Cabaña-Brunod M, Reyes-Ramírez R, Llancalahuen FM, Vilos C y otros. Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines. Journal of Nanobiotechnology. 2017 ene 3;15(1). 1. Disponible desde, DOI: 10.1186/s12951-016-0241-6