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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

Electron microscopy study of SPION interaction and uptake. (A) Electron microscopy images of thin sections of cells interacting with DMSA-SPION by clathrin mediated uptake (<200 nm in diameter aggregates). Scale bar represents 200 nm. (B) Two images by electron microscopy of thin sections of cells showing typical images of macropinocytosis for DMSA-SPION uptake (>200 nm in diameter aggregates). Scale bars represent 200 nm. (C) Electron microscopy images of different types of endosomes containing SPION aggregates: (a) Early endosome. (b) Multivesicular body containing intraluminal vesicles. (c) Late endosome characterized by a multilamellar morphology. (d) Late endosomes and lysosomes with multivesicular structure and large electron-dense areas. Scale bar represents 200 nm.
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Fig4: Electron microscopy study of SPION interaction and uptake. (A) Electron microscopy images of thin sections of cells interacting with DMSA-SPION by clathrin mediated uptake (<200 nm in diameter aggregates). Scale bar represents 200 nm. (B) Two images by electron microscopy of thin sections of cells showing typical images of macropinocytosis for DMSA-SPION uptake (>200 nm in diameter aggregates). Scale bars represent 200 nm. (C) Electron microscopy images of different types of endosomes containing SPION aggregates: (a) Early endosome. (b) Multivesicular body containing intraluminal vesicles. (c) Late endosome characterized by a multilamellar morphology. (d) Late endosomes and lysosomes with multivesicular structure and large electron-dense areas. Scale bar represents 200 nm.

Mentions: To identify the precise mechanism of endocytosis (phagocytosis, pinocytosis, macropinocytosis, clathrin- mediated endocytosis, or caveolae-mediated endocytosis), we performed transmission electron microscopy (TEM) studies. The high contrast of the magnetic particles allowed for their clear identification (Figure 4). Small groups of particles were seen near cell membranes. Actually, SPION incubated in culture media present a relatively wide size distribution (ranging between 50 to more than 400 nm, see Additional file 2). Although we did not make an attempt to sort the SPION by size, we found significant differences in the way the SPION were incorporated in the cells according to the aggregate size. Smaller aggregates were seen adjacent to distinct clathrin-coated patches (Figure 4A). Closed clathrin vesicles containing small DMSA-SPION aggregates (smaller than 200 nm) were seen in the cytoplasm, near membrane. Larger DMSA-SPION aggregates were seen near cell periphery, in most cases engulfed by cell membrane extensions, indicating the existence of a macropinocytic DMSA-SPION uptake process (Figure 4B a, b). Other studies have also proposed a macropinocytic process for cationic iron oxide nanoparticles internalization [30], as well as for other nanoparticles [31].Figure 4


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)

Electron microscopy study of SPION interaction and uptake. (A) Electron microscopy images of thin sections of cells interacting with DMSA-SPION by clathrin mediated uptake (<200 nm in diameter aggregates). Scale bar represents 200 nm. (B) Two images by electron microscopy of thin sections of cells showing typical images of macropinocytosis for DMSA-SPION uptake (>200 nm in diameter aggregates). Scale bars represent 200 nm. (C) Electron microscopy images of different types of endosomes containing SPION aggregates: (a) Early endosome. (b) Multivesicular body containing intraluminal vesicles. (c) Late endosome characterized by a multilamellar morphology. (d) Late endosomes and lysosomes with multivesicular structure and large electron-dense areas. Scale bar represents 200 nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4403785&req=5

Fig4: Electron microscopy study of SPION interaction and uptake. (A) Electron microscopy images of thin sections of cells interacting with DMSA-SPION by clathrin mediated uptake (<200 nm in diameter aggregates). Scale bar represents 200 nm. (B) Two images by electron microscopy of thin sections of cells showing typical images of macropinocytosis for DMSA-SPION uptake (>200 nm in diameter aggregates). Scale bars represent 200 nm. (C) Electron microscopy images of different types of endosomes containing SPION aggregates: (a) Early endosome. (b) Multivesicular body containing intraluminal vesicles. (c) Late endosome characterized by a multilamellar morphology. (d) Late endosomes and lysosomes with multivesicular structure and large electron-dense areas. Scale bar represents 200 nm.
Mentions: To identify the precise mechanism of endocytosis (phagocytosis, pinocytosis, macropinocytosis, clathrin- mediated endocytosis, or caveolae-mediated endocytosis), we performed transmission electron microscopy (TEM) studies. The high contrast of the magnetic particles allowed for their clear identification (Figure 4). Small groups of particles were seen near cell membranes. Actually, SPION incubated in culture media present a relatively wide size distribution (ranging between 50 to more than 400 nm, see Additional file 2). Although we did not make an attempt to sort the SPION by size, we found significant differences in the way the SPION were incorporated in the cells according to the aggregate size. Smaller aggregates were seen adjacent to distinct clathrin-coated patches (Figure 4A). Closed clathrin vesicles containing small DMSA-SPION aggregates (smaller than 200 nm) were seen in the cytoplasm, near membrane. Larger DMSA-SPION aggregates were seen near cell periphery, in most cases engulfed by cell membrane extensions, indicating the existence of a macropinocytic DMSA-SPION uptake process (Figure 4B a, b). Other studies have also proposed a macropinocytic process for cationic iron oxide nanoparticles internalization [30], as well as for other nanoparticles [31].Figure 4

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