Curing the Toxicity of Multi-Walled Carbon Nanotubes through Native Small-molecule Drugs

With the development and application of nanotechnology, large amounts of nanoparticles will be potentially released to the environment and possibly cause many severe health problems. Although the toxicity of nanoparticles has been investigated, prevention and treatment of damages caused by nanoparticles have been rarely studied. Therefore, isotope tracing and improved CT imaging techniques were used to investigate the biodistribution influence between oMWCNTs(oxidized multi-walled carbon nanotubes) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC)/or simvastatin (TD) in vivo. What’s more, biochemical indices in plasma and tissue histology were measured to further study therapeutic effects on the damages of oMWCNTs in mice. Isotope tracing and improved CT imaging results showed that low dosages of DOPC and TD didn’t affect the distribution of oMWCNTs in mice; conversely, the distribution and metabolism of DOPC and TD were influenced by oMWCNTs. Moreover, DOPC and/or TD improved the biocompatibility of oMWCNTs in erythrocyte suspension in vitro. Biochemical index and histopathological results indicated that DOPC and TD didn’t prevent injuries caused by oMWCNTs effectively. But TD showed a good therapeutic effect for damages. This study is the first to investigate prevention and treatment effects of drugs on damages caused by oMWCNTs and provides new insights and breakthroughs for management of nanoparticles on health hazards.

The TEM characterization of Ag/oMWCNTs. Figure S2 The FT-IR characterization of Ag/oMWCNTs and oMWCNTs. Figure S3 The Raman spectrum of Ag/oMWCNTs and oMWCNTs. Figure S4 The labeling yields characterization of 131 I-compounds through paper chromatogram, the chromatographic solutions were normal saline and acetone. Note: In this section, A sample of the test solution (10 μL) was applied at 1 cm from the lower end of the strips (Whatman paper, Maidstone, Kent, UK). The strips were developed in normal saline and acetone until the solvent reached the top portion. The strips were dried and cut into 1-cm segments, and the distribution of radioactivity on the strip was determined using a gamma-ray counter. According to the solubility property of Na 131 I, DOPC/or TD and oMWCNTs in normal saline and acetone, the labeling yields of 131 I-oMWCNTs, 131 I-DOPC and 131 I-TD measured through normal saline were 82.82%, 70.26% and 72.21%, respectively; and the labeling yields of 131 I-DOPC and 131 I-TD measured through acetone were 74.28% and 78.57%, respectively, which were all over 60%, and so the radiotracing technique used in this paper could reflect the biodistribution and behavior of 131 I-compounds effectively in vivo.   single CNTs group; n=5-6, +sem). Figure S8 The morphological alterations in liver, lung, spleen, kidney and heart tissues after exposure different drugs (C for the tissues of oMWCNTs-model group mice, CM-D for the tissues of the tissues of oMWCNTs-model +DOPC group mice, CM-T for the tissues of oMWCNTs-model +TD group mice, DM-C for the tissues of DOPC-model +oMWCNTs group mice, TM-C for the tissues of TD-model +oMWCNTs group mice, D-C for the tissues of DOPC+oMWCNTs group mice, T-C for the tissues of TD+oMWCNTs group mice, respectively. * p<0.05 for groups vs. oMWCNTs-model group; n=5-6, +sem).
Note: In this experiment, the histology score of liver tissue was determined as Anita Patlolla et al [1] reported that 0= normal, 1 = mild cellular disruption in less than 25% of field area, 2 = moderate cellular disruption and hepato cellular vacuolation greater than 50% of field area, 3 = extensive cell disruption, hepato cellular vacuolation and condensed nuclei (pycknotic) of hepatocytes in greater than 50% of field area, 4 = extensive cell disruption, hepato cellular vacuolation, pycknotic and occasional central vein injury and 5= extensive cell disruption, multi central vein necrosis and degenerating of liver in more than 50% of field area. However, the assessment of histology scores of other tissues was determined as the following method: Histology score Present 0 Not present 1 This corresponds to a histologic change that may be barely noticeable to changes considered so minor, small, or infrequent as to warrant no more than the least assignable grade. For focal, multifocal or diffusely distributed lesions, this grade is used for processes where <10% of the tissue is involved. For hyperplastic/hypoplastic/atrophic lesions, this grade is used when the affected structure or tissue has undergone <10% increase or decrease in volume.
2 This corresponds to a histologic change that is a noticeable but not a prominent feature of the tissue. For focal, multifocal or diffusely distributed lesions, this grade is used for processes where between 10-39% of the tissue is involved. For hyperplastic/hypoplastic/atrophic lesions, this grade is used when the affected structure or tissue has undergone between an 10% and 39% increase or decrease in volume.
3 This corresponds to a histologic change that is a prominent feature of the tissue. For focal, multifocal or diffusely distributed lesions, this grade is used for processes where 40-79% of the tissue section is involved. For hyperplastic/hypoplastic/atrophic lesions, this grade is used when the affected structure or tissue has undergone between a 40% and 79% increase or decrease in volume.

4
This corresponds to a histologic change that is an overwhelming feature of the tissue. For focal, multifocal or diffusely distributed lesions, this grade is used for processes where 80-100% of the tissue section is involved. For hyperplastic/hypoplastic/atrophic lesions, this grade is used when the affected structure or tissue has undergone between a 80% and 100% increase or decrease in volume. Figure S9 The level changes of white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB), red blood cell specific volume (HCT), blood platelet (PLT) and plateletcrit (PCT) in blood after exposure 12.4 mg/kg.bw oMWCNTs with 6.2, 12.4, 18.6 mg/kg.bw TD (A) and DOPC (B) to mice (*p<0.05 for groups vs. control group mice; & p<0.05 for groups vs. oMWCNTs-model group; n=4, +sem).
Figure S10 Relative viabilities of 4T1 cells at 24h after incubation with various dosages of oMWCNTs with 2.5 μg/mL TD (*p<0.05 for groups vs. single oMWCNTs group mice, n=10, +sem). (Note: In this experiment, 4T1 cells were incubated with 0, 2, 4, 10, 20 and 40 μg/mL single oMWCNTs or mixture with 2.5 μg/mL TD for 24h, then the relative cell viabilities were measured by the MTT assay, through which to further prove the anti-inflammatory effect of TD on the damage of oMWCNTs in mice.)