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Characterization of interaction of magnetic nanoparticles with breast cancer cells.

Calero M, Chiappi M, Lazaro-Carrillo A, Rodríguez MJ, Chichón FJ, Crosbie-Staunton K, Prina-Mello A, Volkov Y, Villanueva A, Carrascosa JL - J Nanobiotechnology (2015)

Bottom Line: Superparamagnetic iron oxide nanoparticles were internalized by energy dependent endocytosis and localized in endosomes.Transmission electron microscopy studies showed macropinocytosis uptake and clathrin-mediated internalization depending on the nanoparticles aggregate size.MCF-7 cells accumulated these nanoparticles without any significant effect on cell morphology, cytoskeleton organization, cell cycle distribution, reactive oxygen species generation and cell viability, showing a similar behavior to untreated control cells.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain. macarena.calero@uam.es.

ABSTRACT

Background: Different superparamagnetic iron oxide nanoparticles have been tested for their potential use in cancer treatment, as they enter into cells with high effectiveness, do not induce cytotoxicity, and are retained for relatively long periods of time inside the cells. We have analyzed the interaction, internalization and biocompatibility of dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles with an average diameter of 15 nm and negative surface charge in MCF-7 breast cancer cells.

Results: Cells were incubated with dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles for different time intervals, ranging from 0.5 to 72 h. These nanoparticles showed efficient internalization and relatively slow clearance. Time-dependent uptake studies demonstrated the maximum accumulation of dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles after 24 h of incubation, and afterwards they were slowly removed from cells. Superparamagnetic iron oxide nanoparticles were internalized by energy dependent endocytosis and localized in endosomes. Transmission electron microscopy studies showed macropinocytosis uptake and clathrin-mediated internalization depending on the nanoparticles aggregate size. MCF-7 cells accumulated these nanoparticles without any significant effect on cell morphology, cytoskeleton organization, cell cycle distribution, reactive oxygen species generation and cell viability, showing a similar behavior to untreated control cells.

Conclusions: All these findings indicate that dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles have excellent properties in terms of efficiency and biocompatibility for application to target breast cancer cells.

No MeSH data available.


Related in: MedlinePlus

Analysis of cytoskeleton. (A) Representative images of cells immunostained for α-tubulin (green) and DNA counterstained with Hoechst-33258 (blue). (a) Interphase control cells. (a’) Metaphase control cell. (b-b’) Interphase cells incubated for 24 h with DMSA-SPION and observed by fluorescence and bright-field microscopy, respectively; (c-c’) cells incubated for 48 h; (d-d’) cells incubated for 72 h. (e-e’) Mitotic spindle of cells incubated for 24 h, (f-f’) 48 h and (g-g’) 72 h. (B) Merged images of F-actin labeled with rhodamine-phalloidin (red), vinculin immunostaining (green) and DNA counterstained with Hoechst-33258 (blue). (a) Control untreated cells. (b-b’) Cells treated with DMSA-SPION for 24 h and observed by fluorescence and bright-field microscopy, respectively. (c-c’) Cells incubated for 48 h. (d-d’) Cells incubated for 72 h. Scale bar 10 μm.
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Fig6: Analysis of cytoskeleton. (A) Representative images of cells immunostained for α-tubulin (green) and DNA counterstained with Hoechst-33258 (blue). (a) Interphase control cells. (a’) Metaphase control cell. (b-b’) Interphase cells incubated for 24 h with DMSA-SPION and observed by fluorescence and bright-field microscopy, respectively; (c-c’) cells incubated for 48 h; (d-d’) cells incubated for 72 h. (e-e’) Mitotic spindle of cells incubated for 24 h, (f-f’) 48 h and (g-g’) 72 h. (B) Merged images of F-actin labeled with rhodamine-phalloidin (red), vinculin immunostaining (green) and DNA counterstained with Hoechst-33258 (blue). (a) Control untreated cells. (b-b’) Cells treated with DMSA-SPION for 24 h and observed by fluorescence and bright-field microscopy, respectively. (c-c’) Cells incubated for 48 h. (d-d’) Cells incubated for 72 h. Scale bar 10 μm.

Mentions: Two components of cytoskeleton were analyzed: microtubules (MTs) and actin filaments (F-actin). MTs are highly dynamic fibers of the cytoskeleton, with critical functions in eukaryotic cells including intracellular transport, organization of cell structural dynamics and cell division. We have evaluated the effects of nanoparticle internalization on MTs during interphase and mitosis by means of indirect immunofluorescence analysis to α-tubulin (DNA counterstained with Hoechst-33258). Figure 6A shows fluorescence images of MTs (green) and DNA (blue) for interphase and metaphase MCF-7 control cells. After 24, 48 or 72 h of incubation with nanoparticles, interphase microtubules maintain their normal morphology and distribution. In the same samples, DMSA-SPION were visualized inside the cells by bright field microscopy. Distributions of mitotic spindles and chromosomes were also similar to metaphase control cells up to 72 h after incubation.Figure 6


Characterization of interaction of magnetic nanoparticles with breast cancer cells.

Calero M, Chiappi M, Lazaro-Carrillo A, Rodríguez MJ, Chichón FJ, Crosbie-Staunton K, Prina-Mello A, Volkov Y, Villanueva A, Carrascosa JL - J Nanobiotechnology (2015)

Analysis of cytoskeleton. (A) Representative images of cells immunostained for α-tubulin (green) and DNA counterstained with Hoechst-33258 (blue). (a) Interphase control cells. (a’) Metaphase control cell. (b-b’) Interphase cells incubated for 24 h with DMSA-SPION and observed by fluorescence and bright-field microscopy, respectively; (c-c’) cells incubated for 48 h; (d-d’) cells incubated for 72 h. (e-e’) Mitotic spindle of cells incubated for 24 h, (f-f’) 48 h and (g-g’) 72 h. (B) Merged images of F-actin labeled with rhodamine-phalloidin (red), vinculin immunostaining (green) and DNA counterstained with Hoechst-33258 (blue). (a) Control untreated cells. (b-b’) Cells treated with DMSA-SPION for 24 h and observed by fluorescence and bright-field microscopy, respectively. (c-c’) Cells incubated for 48 h. (d-d’) Cells incubated for 72 h. Scale bar 10 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Fig6: Analysis of cytoskeleton. (A) Representative images of cells immunostained for α-tubulin (green) and DNA counterstained with Hoechst-33258 (blue). (a) Interphase control cells. (a’) Metaphase control cell. (b-b’) Interphase cells incubated for 24 h with DMSA-SPION and observed by fluorescence and bright-field microscopy, respectively; (c-c’) cells incubated for 48 h; (d-d’) cells incubated for 72 h. (e-e’) Mitotic spindle of cells incubated for 24 h, (f-f’) 48 h and (g-g’) 72 h. (B) Merged images of F-actin labeled with rhodamine-phalloidin (red), vinculin immunostaining (green) and DNA counterstained with Hoechst-33258 (blue). (a) Control untreated cells. (b-b’) Cells treated with DMSA-SPION for 24 h and observed by fluorescence and bright-field microscopy, respectively. (c-c’) Cells incubated for 48 h. (d-d’) Cells incubated for 72 h. Scale bar 10 μm.
Mentions: Two components of cytoskeleton were analyzed: microtubules (MTs) and actin filaments (F-actin). MTs are highly dynamic fibers of the cytoskeleton, with critical functions in eukaryotic cells including intracellular transport, organization of cell structural dynamics and cell division. We have evaluated the effects of nanoparticle internalization on MTs during interphase and mitosis by means of indirect immunofluorescence analysis to α-tubulin (DNA counterstained with Hoechst-33258). Figure 6A shows fluorescence images of MTs (green) and DNA (blue) for interphase and metaphase MCF-7 control cells. After 24, 48 or 72 h of incubation with nanoparticles, interphase microtubules maintain their normal morphology and distribution. In the same samples, DMSA-SPION were visualized inside the cells by bright field microscopy. Distributions of mitotic spindles and chromosomes were also similar to metaphase control cells up to 72 h after incubation.Figure 6

Bottom Line: Superparamagnetic iron oxide nanoparticles were internalized by energy dependent endocytosis and localized in endosomes.Transmission electron microscopy studies showed macropinocytosis uptake and clathrin-mediated internalization depending on the nanoparticles aggregate size.MCF-7 cells accumulated these nanoparticles without any significant effect on cell morphology, cytoskeleton organization, cell cycle distribution, reactive oxygen species generation and cell viability, showing a similar behavior to untreated control cells.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain. macarena.calero@uam.es.

ABSTRACT

Background: Different superparamagnetic iron oxide nanoparticles have been tested for their potential use in cancer treatment, as they enter into cells with high effectiveness, do not induce cytotoxicity, and are retained for relatively long periods of time inside the cells. We have analyzed the interaction, internalization and biocompatibility of dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles with an average diameter of 15 nm and negative surface charge in MCF-7 breast cancer cells.

Results: Cells were incubated with dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles for different time intervals, ranging from 0.5 to 72 h. These nanoparticles showed efficient internalization and relatively slow clearance. Time-dependent uptake studies demonstrated the maximum accumulation of dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles after 24 h of incubation, and afterwards they were slowly removed from cells. Superparamagnetic iron oxide nanoparticles were internalized by energy dependent endocytosis and localized in endosomes. Transmission electron microscopy studies showed macropinocytosis uptake and clathrin-mediated internalization depending on the nanoparticles aggregate size. MCF-7 cells accumulated these nanoparticles without any significant effect on cell morphology, cytoskeleton organization, cell cycle distribution, reactive oxygen species generation and cell viability, showing a similar behavior to untreated control cells.

Conclusions: All these findings indicate that dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles have excellent properties in terms of efficiency and biocompatibility for application to target breast cancer cells.

No MeSH data available.


Related in: MedlinePlus