Internalization of Titanium Dioxide Nanoparticles Is Mediated by Actin-Dependent Reorganization and Clathrin- and Dynamin-Mediated Endocytosis in H9c2 Rat Cardiomyoblasts

Elizabeth Huerta-García, María Del Pilar Ramos-Godinez, Alejandro López-Saavedra, Ernesto Alfaro-Moreno, Nancy Patricia Gómez-Crisóstomo, Zaira Colín-Val, Helen Sánchez-Barrera, Rebeca López-Marure

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Titanium dioxide nanoparticles (TiO 2 NPs) are widely used for industrial and commercial applications. Once inside the body, they translocate into the bloodstream and reach different areas of the cardiovascular system including the heart, increasing the risk of developing cardiovascular diseases; consequently, the investigation of their interaction with cardiac cells is required. We previously showed that TiO 2 NPs are internalized by H9c2 rat cardiomyoblasts, and here, we examined the molecular mechanisms underlying this process. TiO 2 NPs internalization was evaluated by transmission electron microscopy, time-lapse microscopy, and flow cytometry. Changes in the actin cytoskeleton were studied by phalloidin staining. Endocytic uptake mechanisms for nanoparticles were probed with chemical inhibitors, whereas clathrin and dynamin expression was measured by Western blot. Cellular uptake of TiO 2 NPs occurred early after 30 min exposure, and large aggregates were observed after 1 h. Actin cytoskeleton reorganization included cell elongation plus lower density and stability of actin fibers. Cytochalasin-D inhibited TiO 2 NPs uptake, indicating actin-mediated internalization. Dynamin and clathrin levels increased early after TiO 2 NPs exposure, and their inhibition reduced nanoparticle uptake. Therefore, TiO 2 NPs internalization by H9c2 rat cardiomyoblasts involves actin cytoskeleton reorganization and clathrin/dynamin-mediated endocytosis.

Original languageEnglish (US)
Pages (from-to)578-588
Number of pages11
JournalChemical Research in Toxicology
Volume32
Issue number4
DOIs
StatePublished - Apr 15 2019

Fingerprint

Dynamins
Clathrin
Endocytosis
Actin Cytoskeleton
Nanoparticles
Rats
Actins
Phalloidine
Cytochalasin D
Cardiovascular System
Transmission Electron Microscopy
Microscopy
Flow Cytometry
Cardiovascular Diseases
Cardiovascular system
Western Blotting
Flow cytometry
Staining and Labeling
Elongation
Microscopic examination

ASJC Scopus subject areas

  • Toxicology

Cite this

Internalization of Titanium Dioxide Nanoparticles Is Mediated by Actin-Dependent Reorganization and Clathrin- and Dynamin-Mediated Endocytosis in H9c2 Rat Cardiomyoblasts. / Huerta-García, Elizabeth; Ramos-Godinez, María Del Pilar; López-Saavedra, Alejandro; Alfaro-Moreno, Ernesto; Gómez-Crisóstomo, Nancy Patricia; Colín-Val, Zaira; Sánchez-Barrera, Helen; López-Marure, Rebeca.

In: Chemical Research in Toxicology, Vol. 32, No. 4, 15.04.2019, p. 578-588.

Research output: Contribution to journalArticle

Huerta-García, Elizabeth ; Ramos-Godinez, María Del Pilar ; López-Saavedra, Alejandro ; Alfaro-Moreno, Ernesto ; Gómez-Crisóstomo, Nancy Patricia ; Colín-Val, Zaira ; Sánchez-Barrera, Helen ; López-Marure, Rebeca. / Internalization of Titanium Dioxide Nanoparticles Is Mediated by Actin-Dependent Reorganization and Clathrin- and Dynamin-Mediated Endocytosis in H9c2 Rat Cardiomyoblasts. In: Chemical Research in Toxicology. 2019 ; Vol. 32, No. 4. pp. 578-588.
@article{c3425f46e39148e0b2c9b0ec1b7e340d,
title = "Internalization of Titanium Dioxide Nanoparticles Is Mediated by Actin-Dependent Reorganization and Clathrin- and Dynamin-Mediated Endocytosis in H9c2 Rat Cardiomyoblasts",
abstract = "Titanium dioxide nanoparticles (TiO 2 NPs) are widely used for industrial and commercial applications. Once inside the body, they translocate into the bloodstream and reach different areas of the cardiovascular system including the heart, increasing the risk of developing cardiovascular diseases; consequently, the investigation of their interaction with cardiac cells is required. We previously showed that TiO 2 NPs are internalized by H9c2 rat cardiomyoblasts, and here, we examined the molecular mechanisms underlying this process. TiO 2 NPs internalization was evaluated by transmission electron microscopy, time-lapse microscopy, and flow cytometry. Changes in the actin cytoskeleton were studied by phalloidin staining. Endocytic uptake mechanisms for nanoparticles were probed with chemical inhibitors, whereas clathrin and dynamin expression was measured by Western blot. Cellular uptake of TiO 2 NPs occurred early after 30 min exposure, and large aggregates were observed after 1 h. Actin cytoskeleton reorganization included cell elongation plus lower density and stability of actin fibers. Cytochalasin-D inhibited TiO 2 NPs uptake, indicating actin-mediated internalization. Dynamin and clathrin levels increased early after TiO 2 NPs exposure, and their inhibition reduced nanoparticle uptake. Therefore, TiO 2 NPs internalization by H9c2 rat cardiomyoblasts involves actin cytoskeleton reorganization and clathrin/dynamin-mediated endocytosis.",
author = "Elizabeth Huerta-Garc{\'i}a and Ramos-Godinez, {Mar{\'i}a Del Pilar} and Alejandro L{\'o}pez-Saavedra and Ernesto Alfaro-Moreno and G{\'o}mez-Cris{\'o}stomo, {Nancy Patricia} and Zaira Col{\'i}n-Val and Helen S{\'a}nchez-Barrera and Rebeca L{\'o}pez-Marure",
year = "2019",
month = "4",
day = "15",
doi = "10.1021/acs.chemrestox.8b00284",
language = "English (US)",
volume = "32",
pages = "578--588",
journal = "Chemical Research in Toxicology",
issn = "0893-228X",
publisher = "American Chemical Society",
number = "4",

}

TY - JOUR

T1 - Internalization of Titanium Dioxide Nanoparticles Is Mediated by Actin-Dependent Reorganization and Clathrin- and Dynamin-Mediated Endocytosis in H9c2 Rat Cardiomyoblasts

AU - Huerta-García, Elizabeth

AU - Ramos-Godinez, María Del Pilar

AU - López-Saavedra, Alejandro

AU - Alfaro-Moreno, Ernesto

AU - Gómez-Crisóstomo, Nancy Patricia

AU - Colín-Val, Zaira

AU - Sánchez-Barrera, Helen

AU - López-Marure, Rebeca

PY - 2019/4/15

Y1 - 2019/4/15

N2 - Titanium dioxide nanoparticles (TiO 2 NPs) are widely used for industrial and commercial applications. Once inside the body, they translocate into the bloodstream and reach different areas of the cardiovascular system including the heart, increasing the risk of developing cardiovascular diseases; consequently, the investigation of their interaction with cardiac cells is required. We previously showed that TiO 2 NPs are internalized by H9c2 rat cardiomyoblasts, and here, we examined the molecular mechanisms underlying this process. TiO 2 NPs internalization was evaluated by transmission electron microscopy, time-lapse microscopy, and flow cytometry. Changes in the actin cytoskeleton were studied by phalloidin staining. Endocytic uptake mechanisms for nanoparticles were probed with chemical inhibitors, whereas clathrin and dynamin expression was measured by Western blot. Cellular uptake of TiO 2 NPs occurred early after 30 min exposure, and large aggregates were observed after 1 h. Actin cytoskeleton reorganization included cell elongation plus lower density and stability of actin fibers. Cytochalasin-D inhibited TiO 2 NPs uptake, indicating actin-mediated internalization. Dynamin and clathrin levels increased early after TiO 2 NPs exposure, and their inhibition reduced nanoparticle uptake. Therefore, TiO 2 NPs internalization by H9c2 rat cardiomyoblasts involves actin cytoskeleton reorganization and clathrin/dynamin-mediated endocytosis.

AB - Titanium dioxide nanoparticles (TiO 2 NPs) are widely used for industrial and commercial applications. Once inside the body, they translocate into the bloodstream and reach different areas of the cardiovascular system including the heart, increasing the risk of developing cardiovascular diseases; consequently, the investigation of their interaction with cardiac cells is required. We previously showed that TiO 2 NPs are internalized by H9c2 rat cardiomyoblasts, and here, we examined the molecular mechanisms underlying this process. TiO 2 NPs internalization was evaluated by transmission electron microscopy, time-lapse microscopy, and flow cytometry. Changes in the actin cytoskeleton were studied by phalloidin staining. Endocytic uptake mechanisms for nanoparticles were probed with chemical inhibitors, whereas clathrin and dynamin expression was measured by Western blot. Cellular uptake of TiO 2 NPs occurred early after 30 min exposure, and large aggregates were observed after 1 h. Actin cytoskeleton reorganization included cell elongation plus lower density and stability of actin fibers. Cytochalasin-D inhibited TiO 2 NPs uptake, indicating actin-mediated internalization. Dynamin and clathrin levels increased early after TiO 2 NPs exposure, and their inhibition reduced nanoparticle uptake. Therefore, TiO 2 NPs internalization by H9c2 rat cardiomyoblasts involves actin cytoskeleton reorganization and clathrin/dynamin-mediated endocytosis.

UR - http://www.scopus.com/inward/record.url?scp=85062568137&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85062568137&partnerID=8YFLogxK

U2 - 10.1021/acs.chemrestox.8b00284

DO - 10.1021/acs.chemrestox.8b00284

M3 - Article

C2 - 30730135

AN - SCOPUS:85062568137

VL - 32

SP - 578

EP - 588

JO - Chemical Research in Toxicology

JF - Chemical Research in Toxicology

SN - 0893-228X

IS - 4

ER -