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Cytotoxic effect of poly-dispersed single walled carbon nanotubes on erythrocytes in vitro and in vivo.

Sachar S, Saxena RK - PLoS ONE (2011)

Bottom Line: Single wall Carbon Nanotubes (SWCNTs) are hydrophobic and do not disperse in aqueous solvents.Administration of AF-SWCNTs through intravenous route resulted in a transient anemia as seen by a sharp decline in blood erythrocyte count accompanied with a significant drop in blood haemoglobin level.Administration of AF-SWCNTs through intratracheal administration also showed significant decline in RBC count while administration through other routes (gavage and intra-peritoneal) was not effective.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.

ABSTRACT
Single wall Carbon Nanotubes (SWCNTs) are hydrophobic and do not disperse in aqueous solvents. Acid functionalization of SWCNTs results in attachment of carboxy and sulfonate groups to carbon atoms and the resulting acid functionalized product (AF-SWCNTs) is negatively charged and disperses easily in water and buffers. In the present study, effect of AF-SWCNTs on blood erythrocytes was examined. Incubation of mouse erythrocytes with AF-SWCNTs and not with control SWCNTs, resulted in a dose and time dependent lysis of erythrocyte. Using fluorescence tagged AF-SWCNTs, binding of AF-SWCNTs with erythrocytes could be demonstrated. Confocal microscopy results indicated that AF-SWCNTs could enter the erythrocytes. Treatment with AF-SWCNTs resulted in exposure of hydrophobic patches on erythrocyte membrane that is indicative of membrane damage. A time and dose dependent increase in externalization of phosphatidylserine on erythrocyte membrane bilayer was also found. Administration of AF-SWCNTs through intravenous route resulted in a transient anemia as seen by a sharp decline in blood erythrocyte count accompanied with a significant drop in blood haemoglobin level. Administration of AF-SWCNTs through intratracheal administration also showed significant decline in RBC count while administration through other routes (gavage and intra-peritoneal) was not effective. By using a recently developed technique of a two step in vivo biotinylation of erythrocytes that enables simultaneous enumeration of young (age <10 days) and old (age>40 days) erythrocytes in mouse blood, it was found that the in vivo toxic effect of AF-SWCNTs was more pronounced on older subpopulation of erythrocytes. Subpopulation of old erythrocytes fell after treatment with AF-SWCNTs but recovered by third day after the intravenous administration of AF-SWCNTs. Taken together our results indicate that treatment with AF-SWCNTs results in acute membrane damage and eventual lysis of erythrocytes. Intravenous administration of AF-SWCNTs resulted in a transient anemia in which older erythrocytes are preferably lysed.

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Effect of Control and AF-SWCNT's on erythrocytes in vitro.Erythrocytes isolated from Swiss mice were cultured in RPMI+10%FBS with different concentrations of control and AF- SWCNT preparations. After different time intervals, residual erythrocyte count was determined. Each point represents Mean±SEM. *p<0.01, **p<0.001 as compared to control untreated groups.
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pone-0022032-g001: Effect of Control and AF-SWCNT's on erythrocytes in vitro.Erythrocytes isolated from Swiss mice were cultured in RPMI+10%FBS with different concentrations of control and AF- SWCNT preparations. After different time intervals, residual erythrocyte count was determined. Each point represents Mean±SEM. *p<0.01, **p<0.001 as compared to control untreated groups.

Mentions: The effect of control and acid-functionalized SWCNTs was examined on murine erythrocytes. For this purpose, blood erythrocytes derived from female Swiss or C57BL/6 mice were cultured with control and acid-functionalized SWCNTs. To facilitate the interactions between carbon nanotubes and erythrocytes, the culture tubes were gently rotated (17 RPM) throughout the culture duration. Erythrocyte recoveries at different time points are shown in Figure 1. Treatment with control SWCNTs had no significant effect on the recovery of erythrocytes. A dose and time dependent decline (70 to 90%) in erythrocyte recovery was however seen in cultures treated with AF-SWCNTs. At 50 µg/ml concentration, a significant decline in erythrocyte recovery was seen even at the earliest 4 h time point. At the 24 h time point, erythrocyte recovery in AF-SWCNT treated cultures fell by 80-90%.


Cytotoxic effect of poly-dispersed single walled carbon nanotubes on erythrocytes in vitro and in vivo.

Sachar S, Saxena RK - PLoS ONE (2011)

Effect of Control and AF-SWCNT's on erythrocytes in vitro.Erythrocytes isolated from Swiss mice were cultured in RPMI+10%FBS with different concentrations of control and AF- SWCNT preparations. After different time intervals, residual erythrocyte count was determined. Each point represents Mean±SEM. *p<0.01, **p<0.001 as compared to control untreated groups.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3139600&req=5

pone-0022032-g001: Effect of Control and AF-SWCNT's on erythrocytes in vitro.Erythrocytes isolated from Swiss mice were cultured in RPMI+10%FBS with different concentrations of control and AF- SWCNT preparations. After different time intervals, residual erythrocyte count was determined. Each point represents Mean±SEM. *p<0.01, **p<0.001 as compared to control untreated groups.
Mentions: The effect of control and acid-functionalized SWCNTs was examined on murine erythrocytes. For this purpose, blood erythrocytes derived from female Swiss or C57BL/6 mice were cultured with control and acid-functionalized SWCNTs. To facilitate the interactions between carbon nanotubes and erythrocytes, the culture tubes were gently rotated (17 RPM) throughout the culture duration. Erythrocyte recoveries at different time points are shown in Figure 1. Treatment with control SWCNTs had no significant effect on the recovery of erythrocytes. A dose and time dependent decline (70 to 90%) in erythrocyte recovery was however seen in cultures treated with AF-SWCNTs. At 50 µg/ml concentration, a significant decline in erythrocyte recovery was seen even at the earliest 4 h time point. At the 24 h time point, erythrocyte recovery in AF-SWCNT treated cultures fell by 80-90%.

Bottom Line: Single wall Carbon Nanotubes (SWCNTs) are hydrophobic and do not disperse in aqueous solvents.Administration of AF-SWCNTs through intravenous route resulted in a transient anemia as seen by a sharp decline in blood erythrocyte count accompanied with a significant drop in blood haemoglobin level.Administration of AF-SWCNTs through intratracheal administration also showed significant decline in RBC count while administration through other routes (gavage and intra-peritoneal) was not effective.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.

ABSTRACT
Single wall Carbon Nanotubes (SWCNTs) are hydrophobic and do not disperse in aqueous solvents. Acid functionalization of SWCNTs results in attachment of carboxy and sulfonate groups to carbon atoms and the resulting acid functionalized product (AF-SWCNTs) is negatively charged and disperses easily in water and buffers. In the present study, effect of AF-SWCNTs on blood erythrocytes was examined. Incubation of mouse erythrocytes with AF-SWCNTs and not with control SWCNTs, resulted in a dose and time dependent lysis of erythrocyte. Using fluorescence tagged AF-SWCNTs, binding of AF-SWCNTs with erythrocytes could be demonstrated. Confocal microscopy results indicated that AF-SWCNTs could enter the erythrocytes. Treatment with AF-SWCNTs resulted in exposure of hydrophobic patches on erythrocyte membrane that is indicative of membrane damage. A time and dose dependent increase in externalization of phosphatidylserine on erythrocyte membrane bilayer was also found. Administration of AF-SWCNTs through intravenous route resulted in a transient anemia as seen by a sharp decline in blood erythrocyte count accompanied with a significant drop in blood haemoglobin level. Administration of AF-SWCNTs through intratracheal administration also showed significant decline in RBC count while administration through other routes (gavage and intra-peritoneal) was not effective. By using a recently developed technique of a two step in vivo biotinylation of erythrocytes that enables simultaneous enumeration of young (age <10 days) and old (age>40 days) erythrocytes in mouse blood, it was found that the in vivo toxic effect of AF-SWCNTs was more pronounced on older subpopulation of erythrocytes. Subpopulation of old erythrocytes fell after treatment with AF-SWCNTs but recovered by third day after the intravenous administration of AF-SWCNTs. Taken together our results indicate that treatment with AF-SWCNTs results in acute membrane damage and eventual lysis of erythrocytes. Intravenous administration of AF-SWCNTs resulted in a transient anemia in which older erythrocytes are preferably lysed.

Show MeSH
Related in: MedlinePlus