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
N1 - Funding Information:
This work was supported by UNAB Regular Grant DI‑316‑13/R. VMM and FDGN thank CONICYT for PhD Scholarship, CONICYT + PAI “Concurso Nacional Tesis de Doctorado en la Empresa”2014 Folio 781413007, Fraunhofer Chile Research, Innova‑Chile CORFO (FCR‑CSB 09CEII‑6991) and Anillo Científico ACT1107. CV acknowledges support from BASAL Grant FB0807, FONDECYT Regular number 1161438, MECESUP PMI‑UAB1301, and UNAB Regular Grant DI‑695. C.O acknowledges to Proyecto Núcleo UNAB DI‑1419‑16/N.
Publisher Copyright:
© 2017 The Author(s).
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.
UR - http://www.scopus.com/inward/record.url?scp=85010209721&partnerID=8YFLogxK
U2 - 10.1186/s12951-016-0241-6
DO - 10.1186/s12951-016-0241-6
M3 - Article
C2 - 28049488
AN - SCOPUS:85010209721
SN - 1477-3155
VL - 15
JO - Journal of Nanobiotechnology
JF - Journal of Nanobiotechnology
IS - 1
M1 - 1
ER -