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In Vitro Neurotoxicity Resulting from Exposure of Cultured Neural Cells to Several Types of Nanoparticles

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ABSTRACT

Laboratory and industrial production of various nanoparticles, single-walled nanotubes (SWNTs), fullerene (C60), cadmium selenide (CdSe) quantum dots, carbon black (CB), and dye-doped silica nanospheres (NSs), has greatly increased in the past 15 years. However, little research has been done to analyze the toxicity of these materials. With recent studies showing that nano-substances can cross the blood–brain barrier, we examined the neurotoxicity of these manufactured nanoparticles. By employing the rat PC-12 neuronal-like cell line as the basis for our studies, we were able to evaluate the toxicity caused by these five nanoparticles. The level of toxicity was measured by testing for cell viability using the lactate dehydrogenase (LDH) cell viability assay, morphological analysis of changes in cellular structures, and Western blot analyses of αII-spectrin breakdown products (SBDP) as cell death indicators. Our results showed cytotoxicity in nondifferentiated PC-12 cells exposed to CB (10–100 μg/mL), SWNTs (10–100 μg/mL), C60 (100 μg/mL), CdSe (10 μg/mL), CB (500 μg/mL), and dye-doped silicon NSs (10 μg/mL). Exposure to higher concentrations (100 μg/mL) of SWNTs, CB, and C60 increased the formation of SBDP150/145, as well as cell membrane contraction and the formation of cytosolic vacuoles. The incorporations of the nanoparticles into cell cytoplasm were observed using the fluorescent dye-doped NSs in both nondifferentiated and nerve growth factor (NGF)-differentiated PC-12 cells. When PC-12 cells are differentiated, they appeared to be even more sensitive to cytotoxicity of nanoparticles such as CB 10 nm (10–100 μg/mL), CB 100 nm (10–100 μg/mL), and CdSe (1–10 μg/mL).

No MeSH data available.


Cytotoxic effects of various nanoparticles on NGF-differentiated PC-12 cells. LDH release levels after 24-hour incubation with carbon black (5 and 100 nm diameter) and cadmium selenide NPs were significantly higher (at both 1 and 10 μg/mL) than control NGF-differentiated PC-12 cells (Student’s t-test, *P < 0.01).
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f6-10.1177_1179670717694523: Cytotoxic effects of various nanoparticles on NGF-differentiated PC-12 cells. LDH release levels after 24-hour incubation with carbon black (5 and 100 nm diameter) and cadmium selenide NPs were significantly higher (at both 1 and 10 μg/mL) than control NGF-differentiated PC-12 cells (Student’s t-test, *P < 0.01).

Mentions: Finally, to examine the effects in the more neuronal-like NGF-differentiated PC-12 cells, the cells were exposed to a 24-hour incubation with CB (5 nm (1–10 μg/mL) and 100 nm diameter (1–10 μg/mL)) and CdSe NP (1–10 μg/mL). We found that at all concentrations, all three NPs appear to be cytotoxic to NGF-differentiated PC-12 cells, based on LDH release, and apparent dose responses were not observed (Figure 6).


In Vitro Neurotoxicity Resulting from Exposure of Cultured Neural Cells to Several Types of Nanoparticles
Cytotoxic effects of various nanoparticles on NGF-differentiated PC-12 cells. LDH release levels after 24-hour incubation with carbon black (5 and 100 nm diameter) and cadmium selenide NPs were significantly higher (at both 1 and 10 μg/mL) than control NGF-differentiated PC-12 cells (Student’s t-test, *P < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5392047&req=5

f6-10.1177_1179670717694523: Cytotoxic effects of various nanoparticles on NGF-differentiated PC-12 cells. LDH release levels after 24-hour incubation with carbon black (5 and 100 nm diameter) and cadmium selenide NPs were significantly higher (at both 1 and 10 μg/mL) than control NGF-differentiated PC-12 cells (Student’s t-test, *P < 0.01).
Mentions: Finally, to examine the effects in the more neuronal-like NGF-differentiated PC-12 cells, the cells were exposed to a 24-hour incubation with CB (5 nm (1–10 μg/mL) and 100 nm diameter (1–10 μg/mL)) and CdSe NP (1–10 μg/mL). We found that at all concentrations, all three NPs appear to be cytotoxic to NGF-differentiated PC-12 cells, based on LDH release, and apparent dose responses were not observed (Figure 6).

View Article: PubMed Central - PubMed

ABSTRACT

Laboratory and industrial production of various nanoparticles, single-walled nanotubes (SWNTs), fullerene (C60), cadmium selenide (CdSe) quantum dots, carbon black (CB), and dye-doped silica nanospheres (NSs), has greatly increased in the past 15 years. However, little research has been done to analyze the toxicity of these materials. With recent studies showing that nano-substances can cross the blood&ndash;brain barrier, we examined the neurotoxicity of these manufactured nanoparticles. By employing the rat PC-12 neuronal-like cell line as the basis for our studies, we were able to evaluate the toxicity caused by these five nanoparticles. The level of toxicity was measured by testing for cell viability using the lactate dehydrogenase (LDH) cell viability assay, morphological analysis of changes in cellular structures, and Western blot analyses of &alpha;II-spectrin breakdown products (SBDP) as cell death indicators. Our results showed cytotoxicity in nondifferentiated PC-12 cells exposed to CB (10&ndash;100 &mu;g/mL), SWNTs (10&ndash;100 &mu;g/mL), C60 (100 &mu;g/mL), CdSe (10 &mu;g/mL), CB (500 &mu;g/mL), and dye-doped silicon NSs (10 &mu;g/mL). Exposure to higher concentrations (100 &mu;g/mL) of SWNTs, CB, and C60 increased the formation of SBDP150/145, as well as cell membrane contraction and the formation of cytosolic vacuoles. The incorporations of the nanoparticles into cell cytoplasm were observed using the fluorescent dye-doped NSs in both nondifferentiated and nerve growth factor (NGF)-differentiated PC-12 cells. When PC-12 cells are differentiated, they appeared to be even more sensitive to cytotoxicity of nanoparticles such as CB 10 nm (10&ndash;100 &mu;g/mL), CB 100 nm (10&ndash;100 &mu;g/mL), and CdSe (1&ndash;10 &mu;g/mL).

No MeSH data available.