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High dispersity of carbon nanotubes diminishes immunotoxicity in spleen.

Lee S, Khang D, Kim SH - Int J Nanomedicine (2015)

Bottom Line: For cytotoxicity of swCNTs, MTT assay, reactive oxygen species production, superoxide dismutase activity, cellular uptake, and confocal microscopy were used in macrophages.In short, less-dispersed swCNTs caused cytotoxicity in macrophages and abnormalities in immune organs such as spleen, whereas highly dispersed swCNTs did not result in immunotoxicity.Our findings clarified the effective immunotoxicological factors of swCNTs by increasing dispersity of swCNTs and provided useful guidelines for the effective use of nanomaterials.

View Article: PubMed Central - PubMed

Affiliation: CMRI, Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.

ABSTRACT

Background: From the various physiochemical material properties, the chemical functionalization order of single-walled carbon nanotubes (swCNTs) has not been considered as a critical factor for modulating immunological responses and toxicological aspects in drug delivery applications. Although most nanomaterials, including carbon nanotubes, are specifically accumulated in spleen, few studies have focused on spleen immunotoxicity. For this reason, this study demonstrated that the dispersity of swCNTs significantly influenced immunotoxicity in vitro and in vivo.

Materials and methods: For cytotoxicity of swCNTs, MTT assay, reactive oxygen species production, superoxide dismutase activity, cellular uptake, and confocal microscopy were used in macrophages. In the in vivo study, female BALB/c mice were intravenously administered with 1 mg/kg/day of swCNTs for 2 weeks. The body weight, organ weight, hematological change, reverse-transcription polymerase chain reaction, and lymphocyte population were evaluated.

Results: Different orders of chemical functionalization of swCNTs controlled immunotoxicity. In short, less-dispersed swCNTs caused cytotoxicity in macrophages and abnormalities in immune organs such as spleen, whereas highly dispersed swCNTs did not result in immunotoxicity.

Conclusion: This study clarified that increasing carboxyl groups on swCNTs significantly mitigated immunotoxicity in vitro and in vivo. Our findings clarified the effective immunotoxicological factors of swCNTs by increasing dispersity of swCNTs and provided useful guidelines for the effective use of nanomaterials.

No MeSH data available.


Related in: MedlinePlus

Effects of dispersed single-walled carbon nanotubes on NO production and cytokine expression.Notes: (A) Cells (2×104 cells/well in 96-well plates) were treated with 1 μg/mL of differently dispersed swCNTs. After treatment for 48 hours, NO production in the culture media was assayed using Griess reagent. Lipopolysaccharide (30 ng/mL) was used as a positive control. (B, C) Cells (2×105 cells/well in 24-well plates) were treated with 1 μg/mL swCNTs for 24 hours. Gene expression of (B) IL-1β and (C) TNF-α was analyzed using real-time PCR. The results are presented as mean ± SE of three independent experiments. *P<0.05 significantly different from control.Abbreviations: CON, control; IL, interleukin; LPS, lipopolysaccharide; PCR, polymerase chain reaction; SE, standard error; swCNT, single-walled carbon nanotube; TNF, tumor necrosis factor.
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f4-ijn-10-2697: Effects of dispersed single-walled carbon nanotubes on NO production and cytokine expression.Notes: (A) Cells (2×104 cells/well in 96-well plates) were treated with 1 μg/mL of differently dispersed swCNTs. After treatment for 48 hours, NO production in the culture media was assayed using Griess reagent. Lipopolysaccharide (30 ng/mL) was used as a positive control. (B, C) Cells (2×105 cells/well in 24-well plates) were treated with 1 μg/mL swCNTs for 24 hours. Gene expression of (B) IL-1β and (C) TNF-α was analyzed using real-time PCR. The results are presented as mean ± SE of three independent experiments. *P<0.05 significantly different from control.Abbreviations: CON, control; IL, interleukin; LPS, lipopolysaccharide; PCR, polymerase chain reaction; SE, standard error; swCNT, single-walled carbon nanotube; TNF, tumor necrosis factor.

Mentions: Although each dispersed swCNT (COOH-low, -mid, and -high) was similarly translocated into the cytoplasm after cellular uptake (Figure 2C), COOH-max particles induced lower NO production and expression of proinflammatory cytokines such as IL-1β and TNF-α (Figure 4A–C). These results suggest that different degrees of swCNT dispersion cause different toxicity and inflammatory responses, although all degrees of swCNT dispersion resulted in similar cellular uptake.


High dispersity of carbon nanotubes diminishes immunotoxicity in spleen.

Lee S, Khang D, Kim SH - Int J Nanomedicine (2015)

Effects of dispersed single-walled carbon nanotubes on NO production and cytokine expression.Notes: (A) Cells (2×104 cells/well in 96-well plates) were treated with 1 μg/mL of differently dispersed swCNTs. After treatment for 48 hours, NO production in the culture media was assayed using Griess reagent. Lipopolysaccharide (30 ng/mL) was used as a positive control. (B, C) Cells (2×105 cells/well in 24-well plates) were treated with 1 μg/mL swCNTs for 24 hours. Gene expression of (B) IL-1β and (C) TNF-α was analyzed using real-time PCR. The results are presented as mean ± SE of three independent experiments. *P<0.05 significantly different from control.Abbreviations: CON, control; IL, interleukin; LPS, lipopolysaccharide; PCR, polymerase chain reaction; SE, standard error; swCNT, single-walled carbon nanotube; TNF, tumor necrosis factor.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4388092&req=5

f4-ijn-10-2697: Effects of dispersed single-walled carbon nanotubes on NO production and cytokine expression.Notes: (A) Cells (2×104 cells/well in 96-well plates) were treated with 1 μg/mL of differently dispersed swCNTs. After treatment for 48 hours, NO production in the culture media was assayed using Griess reagent. Lipopolysaccharide (30 ng/mL) was used as a positive control. (B, C) Cells (2×105 cells/well in 24-well plates) were treated with 1 μg/mL swCNTs for 24 hours. Gene expression of (B) IL-1β and (C) TNF-α was analyzed using real-time PCR. The results are presented as mean ± SE of three independent experiments. *P<0.05 significantly different from control.Abbreviations: CON, control; IL, interleukin; LPS, lipopolysaccharide; PCR, polymerase chain reaction; SE, standard error; swCNT, single-walled carbon nanotube; TNF, tumor necrosis factor.
Mentions: Although each dispersed swCNT (COOH-low, -mid, and -high) was similarly translocated into the cytoplasm after cellular uptake (Figure 2C), COOH-max particles induced lower NO production and expression of proinflammatory cytokines such as IL-1β and TNF-α (Figure 4A–C). These results suggest that different degrees of swCNT dispersion cause different toxicity and inflammatory responses, although all degrees of swCNT dispersion resulted in similar cellular uptake.

Bottom Line: For cytotoxicity of swCNTs, MTT assay, reactive oxygen species production, superoxide dismutase activity, cellular uptake, and confocal microscopy were used in macrophages.In short, less-dispersed swCNTs caused cytotoxicity in macrophages and abnormalities in immune organs such as spleen, whereas highly dispersed swCNTs did not result in immunotoxicity.Our findings clarified the effective immunotoxicological factors of swCNTs by increasing dispersity of swCNTs and provided useful guidelines for the effective use of nanomaterials.

View Article: PubMed Central - PubMed

Affiliation: CMRI, Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.

ABSTRACT

Background: From the various physiochemical material properties, the chemical functionalization order of single-walled carbon nanotubes (swCNTs) has not been considered as a critical factor for modulating immunological responses and toxicological aspects in drug delivery applications. Although most nanomaterials, including carbon nanotubes, are specifically accumulated in spleen, few studies have focused on spleen immunotoxicity. For this reason, this study demonstrated that the dispersity of swCNTs significantly influenced immunotoxicity in vitro and in vivo.

Materials and methods: For cytotoxicity of swCNTs, MTT assay, reactive oxygen species production, superoxide dismutase activity, cellular uptake, and confocal microscopy were used in macrophages. In the in vivo study, female BALB/c mice were intravenously administered with 1 mg/kg/day of swCNTs for 2 weeks. The body weight, organ weight, hematological change, reverse-transcription polymerase chain reaction, and lymphocyte population were evaluated.

Results: Different orders of chemical functionalization of swCNTs controlled immunotoxicity. In short, less-dispersed swCNTs caused cytotoxicity in macrophages and abnormalities in immune organs such as spleen, whereas highly dispersed swCNTs did not result in immunotoxicity.

Conclusion: This study clarified that increasing carboxyl groups on swCNTs significantly mitigated immunotoxicity in vitro and in vivo. Our findings clarified the effective immunotoxicological factors of swCNTs by increasing dispersity of swCNTs and provided useful guidelines for the effective use of nanomaterials.

No MeSH data available.


Related in: MedlinePlus