Toxicity of poly-dispersed single-walled carbon nanotubes on bone marrow derived Hematopoietic Stem and Progenitor Cells.

This study has explored the effect of acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) on Hematopoietic Stem and Progenitor Cell (HSPCs) in mouse bone marrow. Administration of AF-SWCNTs induced a significant decline in the live-cell recovery from bone marrow. Lin-negative Stem cell enriched HSPCs internalized AF-SWCNTs that remained localized in cytoplasmic areas. Incubation of HSPCs with AF-SWCNTs resulted in induction of cell death, inhibition of cell cycle, and induction of reactive oxygen species (ROS) as well as the expression of Caspase 3, 7 and 9 enzymes. In vitro culture with a cytokine cocktail (SCF, GM-CSF, IL3, IL6, IL7) induced differentiation of HSPCs into lymphocytes and myeloid cells, that was inhibited in presence of AF-SWCNTs. Relative recoveries of lymphocytes specifically B lymphocytes, was significantly reduced by AF-SWCNT-treatment, whereas the relative recovery of myeloid cells remained unaltered. These results suggest that AF-SWCNTs have significant toxic effects on HSPCs and differentially suppress the ontogeny of lymphoid and myeloid cells.

Poly dispersed acid-functionalized single walled carbon nanotubes target activated T and B cells to suppress acute and chronic GVHD in mouse model.

Graft versus host disease (GVHD) results from hyper-activation of transplanted lymphocytes against the host antigens. Bone marrow transplantation in humans as well as some cases of blood transfusion and organ transplantation are associated with a strong GVH reaction resulting in GVHD that in many cases may be fatal. We had previously shown that poly-dispersed acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) specifically target activated T and B lymphocytes and kill them. In the present study, efficacy of AF-SWCNTs to suppress the GVH reaction was tested in the mouse model. Acute GVHD was induced in mice by administering intravenously 30 or 60 million spleen cells from a parental strain (C57bl/6 mouse, MHC haplotype H-2b) to host (C57bl/6 x Balb/c) F1 mice (MHC haplotype H-2b/d)and waiting for 8-10 days. Chronic GVHD was similarly induced by administration of 30 million parent spleen cells to F1 mice and waiting for a period of 60 days. Our results demonstrate a marked decline in splenomegaly and recovery of spleen T (both CD4 and CD8) and B cells in GVHD mice treated with AF-SWCNTs. AF-SWCNTs treatment also limited T and B cell proliferation by restricting S-phage of cell cycle. Generation of anti-host cytotoxic T cells (CTLs) was also markedly suppressed by AF-SWCNT treatment of acute GVHD mice, and a significant reduction in the generation of anti-host antibodies could also be demonstrated. Taken together, our results suggest that the AF-SWCNTs can be considered as a potential therapeutic agent for treating GVHD.

Acid-functionalized single-walled carbon nanotubes alter epithelial tight junctions and enhance paracellular permeability.

Due to their unique properties, carbon nanotubes (CNTs) are being widely explored for industrial and medical applications. This has necessitated a thorough assessment of the effect of CNTs on human and animal physiology and health. Impact of CNTs on epithelial tight junctions has not been evaluated in the context of their toxic effects in many biological systems. In the present study, we examined the effect of acid functionalized single-walled carbon nanotubes (AF-SWCNTs) on the function and expression of two tight junction proteins (ZO-1 and occludin) in the Madin-Darby canine kidney (MDCK) cell line. Treatment of MDCK cells with AF-SWCNT resulted in a downregulation of tight junction proteins, decreased trans-epithelial electrical resistance (TER), increased paracellular permeability, and disruption of tight junctions. Taken together, our data demonstrate that AF-SWCNT disrupts tight junction barrier by downregulating tight junction proteins in MDCK epithelial cells.

Elevated internalization and cytotoxicity of polydispersed single-walled carbon nanotubes in activated B cells can be basis for preferential depletion of activated B cells in vivo.

Uptake of polydispersed acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) in resting and LPS-activated B cells was studied using fluorescence-tagged AF-SWCNTs (FAF-SWCNTs). Activated B cells internalized substantially higher amounts of FAF-SWCNTs [76.5% AF-SWCNT+ B cells, mean fluorescence intensity (MFI) 720.6] as compared to the resting B cells [39.5% AF-SWCNT+ B cells, MFI 198.5]. B cells in S and G2/M phases were found to have significantly higher uptake of FAF-SWCNTs as compared to cells in G0/G1 phase. Confocal microscopy indicated that AF-SWCNTs were essentially localized on cell membrane in resting B cells, whereas in activated B cells, AF-SWCNTs were distributed throughout the cytoplasm. Targeting of AF-SWCNTs specifically to activated B cells in vivo was examined by first administering intravenously LPS-activated B cells tagged with fluorescence tracer (CFSE) in mice, followed by FAF-SWCNTs through the same route. It was found that FAF-SWCNTs were specifically taken up by CFSE+CD19+-activated B cells (95% FAF-SWCNT+ B cells, MFI 3725) as compared to CFSE- CD19+ resting B cells (31.1% FAF-SWCNT+ B cells, MFI 428). Administration (i.v.) of LPS resulted in a significant increase in the proportion of B cell in mouse spleen that was reduced by 68% by administering AF-SWCNTs. In control mice, the corresponding decrease in B cell proportion was 49%, which was significantly lower (p < 0.005) than the decline in LPS-treated mice. These results indicate that AF-SWCNTs may have the potential as an agent for depleting activated B cells in vivo.