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Table 2 In vivo studies on immunological properties of carbon-based nanomaterials

From: Cellular Toxicity and Immunological Effects of Carbon-based Nanomaterials

Test materials Exposure pathways Exposure doses Suspension buffer Time points of sacrifice Test animals Main findings Refe-rence
MWCNTs Pharyngeal aspiration 40 μg/mouse PBS with 0.6 mg/mL serum albumin and 0.01 mg/mL 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DSPC) 24 h, 7 days, and 28 days C57BL/6 mice aged 10 weeks An early increase in serum cytokines and inflammatory gene expression in serum; consistently increased eosinophils in blood and BALF. [34]
Biodegraded and non-biodegraded carbon nanotubes Pharyngeal aspiration 40 μg/mouse PBS for non-biodegraded CNTs; PBS with hMPO and H2O2 for biodegrated CNTs 7 days C57BL/6 mice No pro-inflammatory pulmonary response observed in mice treated with biodegraded nanotubes. [73]
Carbon black nanoparticles Intratracheal instillation (single) 0.018, 0.054 or 0.162 mg/mouse 0.9% NaCl
MilliQ water with 10% (v/v) acellular BAL from
C57BL/6 mice.
1, 3 and 28 days Female C57BL/6 mice aged 5-6 weeks Strongest lung inflammation on day 1 and day 3 post exposure, elevating for the two highest doses (0.054 and 0.162 mg) 28 days post-exposure. [114]
High-purity WMCNTs Orotracheal aspiration 4 mg/kg body weight PBS with 1% Pluronic F127 1 h, 6 h, 12 h, 18 h, 24 h, 48 h and 72 h CF-1 Non-Swiss Albino mice aged 6 weeks Time-dependent neutrophil influx and pro-inflammatory mediator (TNF-α and IL-6) release in bronchoalveolar lavage (BAL) fluid; attenuated influx of neutrophils in AM-depleted mice. [115]
MWCNTs Intratracheal instillation; inhalation 0.2 mg or 1 mg/mouse; 0.37mg/m3 aerosols (6 h/day, 5 days/week) for 4 weeks Distilled water with 0.05% Triton X 3 days, 1 week, 1 month, 3 months, and 6 months;
3 days, 1 month, and 3 months
Male Wistar rats aged 9 weeks Less amounts of MWCNTs delivered into the lungs, and therefore less pulmonary inflammation responses in the group of inhalation exposure compared to intratracheal instillation. [116]
SWCNTs Intratracheal instillation 0.04, 0.2, 1 mg/kg body weight PBS with 10 mg/mL Tween 80 3 days, 1 week, 1 month, 3 months and 6 months Male Crl: CD (SD) rats aged seven weeks Increased inflammatory cells in BALF in a dose-dependent manner from 3 days post-exposure up to 3 months; alveolar macrophage accumulation and inflammatory cell infiltration in the lung sections. [117]
Long tangled MWCNTs and long rod-like MWCNTs Pharyngeal aspiration 10 or 40 μg/mouse PBS with 0.6 mg/ml BSA 4 and 16 h or 7, 14, and 28 days Female C57BL/6 mice aged 7–8 weeks Attenuated inflammatory reactions caused by CNTs in IL-1R-/-mice and antagonist-treated (etanercept and anakinra) mice [119]
MWCNTs Pharyngeal aspiration 5, 20, or
40 μg/mouse
Ca2+ and Mg2+-free PBS with 0.6 mg/ml mouse serum albumin and 0.01 mg/ml DPPC 1, 3, 7 and 14 days Male C57BL/6J mice aged 8 weeks Rapid and prominent fibrosis formation remarkably near where the particles were deposited in the lungs; pronounced infiltration of neutrophils and macrophages alongside fibrosis. [122]
SWCNTs and fullerenes Intratracheal instillation Low dose: 0.0003, 0.0015, 0.003, 0.015, and 0.3 mg/mouse; high dose: 0.1 and 0.5 mg/mouse Pluronic F-68 7 days Male ICR mice aged 5-6 weeks Airway hyperreactivity and airflow obstruction; upregulation of cathepsin K, MMP-12, CCL2 and CCL3, and macrophage receptors such as Toll-like receptor 2 and macrophage scavenger receptor 1. [123]
SWCNTs, MWCNTs and ultrafine carbon black particles Subcutaneous injection into the footpad together with OVA;
intranasal administration together with OVA.
For the injection model, ~200 μg /single dose (max dose with fourfold dilutions) for 3 doses; for the intranasal model, 133 μg/day for 3consecutive days Hank’s balanced salt
solution with 10% serum from BALB/cA mouse
An OVA booster on days 21, sacrificed on days 26; An OVA booster on days 21,22 and 23, sacrificed on days 26. Female inbred BALB/cAnNCrl mice aged 6–7 weeks Increased serum levels of OVA-specific IgE, number of eosinophils in BALF, and secretion of Th2-associated cytokines in the mediastinal lymph node in the group of CNTs together with OVA; increased IgG2a levels, neutrophil cell numbers, and levels of TNF-α and MCP-1 in BALF in the group of MWCNT and ufCB with OVA. [124]
MWCNTs Intratracheal instillation 50 μg/mouse once a week for 6 weeks PBS with 0.05% Tween 80 24 h after the final intratracheal administration Male ICR mice aged 6-7 weeks Aggravated airway inflammation observed in the group of OVA+MWCNTs, characterized by infiltration of immune cells and production of cytokines; increased levels of serum immunoglobulin (allergen-specific IgG1 and IgE) compared with OVA alone. [125]
Carbon nanoparticles Pharyngeal
2.5 mg/kg body weight PBS 12 h, 24 h, 48 h Female Balb/cJRj mice aged 8 weeks Increased allergic airway inflammation and specific Th2 response in the lymph nodes to OVA in the presence of carbon nanoparticles. [126]
Carbon nanoparticles Intratracheal instillation 20 μg/mouse Pyrogene-free
distilled water
3, 6, 12, 18, 24 h, and 3 or 7 days Female C57BL/6 J mice aged 8–10 weeks Prominent culmination of neutrophil granulocytes 12 to 24 h after instillation; BAL concentrations and increased levels of neutrophil chemoattractants (CXCL1, -2 and-5) from alveolar epithelial type II cells. [127]
MWCNTs Intranasal instillation administered on days 0, 7, and 14 (75 μg/dose) A synthetic lung surfactant On the day 23 Male BALB/cByJ mice aged 9 weeks Aggravated airway inflammation and the generation of epithelium-derived innate cytokines caused by the coexposure of MWCNTs and HDM compared to HDM alone. [128]
Rod-like MWCNTs and tangled MWCNTs Inhalation 6.2-8.2 mg/m3 for rCNT and 17.5-18.5 mg/m3 for tCNT (4 hours at a time once or on four consecutive days) 4 h, 24 h Female C57BL/6 and BALB/c mice aged 7–8 weeks Allergic-like airway inflammation and the upregulation of innate immunity-relevant genes and cytokine/chemokine pathways caused by rod-like CNTs after 4-hour exposure. [129]
MWCNTs Oropharyngeal aspiration 4 mg/kg body weight Saline with 10% surfactant 1 or 30 days C57BL/6J and B6.Cg-kit (W-sh) mast cell deficient mice aged 4–10 weeks Involvement of mast cells and the
IL-33/ST2 axis in the pulmonary and cardiovascular responses to MWCNT exposure; no toxicological effects observed in mast cell-deficient mice or in mice without response to IL-33.
MWCNTs Intratracheal instillation 5, 20, and 50 mg/kg body weight PBS 1,3, 7,14 days Male ICR mice Increased pro-inflammatory cytokines in BALF and in blood in a dose-dependent manner, peaking at day 1 postexposure; more pronounced elevation of Th2-type cytokines than that of Th1-type cytokines; considerably enhanced number of B cells in the spleen and blood. [133]
MWCNTs Intravenous injection 100 μg/mouse Ca2+ and Mg2+-free PBS 24 h, 7 days and 15 days Female C57BL/6 mice aged 6-8 weeks Proliferative response of T lymphocytes to a nonspecific mitogen and to ovalbumin (OVA); an increase in expression of proinflammatory cytokines such as TNF-α and IL-6 and IFN-γ, and a decrease in TGF-β and IL-10; increased antibody production to OVA in the treated group. [134]
nonPEGylated SWCNTs Intravenous injection 1.2 mg/kg body weight PBS 6 min and
60 min
Male Wistar rats of body weight 250–280 g A significant rise in plasma thromboxane B2 levels. [145]
MWCNTs Inhalation 0.3, 1, or 5 mg/m3 (6 h/day) for 7 or 14 days 7 or 14 days Male C57BL/6 mice aged 10 weeks An absence of severe inflammation and tissue injury; decreased NK cell function and T cell-dependent antibody response. [148]
MWCNTs Inhalation 0, 0.3 or 1 mg/m3 (6 h/day) for 14 consecutive days Male C57Bl/6 mice aged approximately 8 weeks A dose-dependent decrease in antibody formation in response to antigen, not altering lymphocyte subpopulations; activation of the cyclooxygenase pathway in the spleen by MWCNTs through TGF-ß release in the lung, leading to T-cell dysfunction and decreased T-cell-dependent antibody formation. [149]
MWCNTs Pharyngeal aspiration 50 μg/mouse PBS with 0.6 mg/ml mouse serum albumin and 0.01 mg/ml DPPC day 7, 28 Wild-type C57BL/6 and IL-1R-/- mice aged 2 months Severe acute pulmonary inflammation, and increases of TNF-α, IL-6, IL-1β and MCP-1 protein levels in BALF in wild-type mice; reduced pulmonary inflammatory response in IL-1R-/- mice exposed to MWCNT. [157]
Four samples of MWCNTs: NTtang1 (diameter ~14.84 nm, length 1-5 μm), NTtang2 (diameter ~10.40 nm, length 5-20 μm), NTlong1 (diameter ~84.89 nm, mean length 13 μm) and NTlong2 (diameter ~165.02 nm, max length 56 μm) Intraperitoneal injection 50 μg/mouse Saline with 0.5% BSA 24 h and 7 days Female C57BL/6 mice aged 8 weeks Significant polymorphonuclear leukocyte (PMN) or protein exudation and granulomas formation on the peritoneal side of the diaphragm in the group of long-fiber-containing samples. [160]
Nano-sized carbon black Inhalation For a 3-day experiment, 13.08 ± 3.18 mg/m3 of non-sonicated carbon black (group N) and 13.67 ± 3.54 mg/m3 of sonicated carbon black (group S); for a 2-week experiment, 9.83 ±3.42 mg/m3 of non-sonicated carbon black and 9.08 ± 4.49 mg/m3 of sonicated carbon black. (exposed 6 h/day, 5 days/week for 3 days or 2 weeks) Distilled water Male Sprague-Dawley (SD) rats aged 5 weeks More carbon black particles-laden macrophages observed in BALF and more carbon black deposited in the lungs exposed to sonicated carbon black; no significant difference in the levels of inflammatory cytokines or damage-indicating proteins between the two groups in the 3-day experiment, whereas 2-week exposure induced increased number of total cells, macrophages, and PMNs in the group S. [163]
SWCNTs, MWCNTs (diameter 10-15 nm), MWCNTs (diameter 20-50 nm) and MWCNTs (diameter 3-10 nm) Intratracheal instillation 100 mg/rat NaCl solution with or without 0.5 mg/ml BSA 24 h Male Sprague Dawley rats of body weight 180-220 g Remarkably accentuated inflammatory cell infiltration in BALF, and increased the number of CNT-loaded alveolar macrophages, caused by CNTs dispersed in BSA, but not in NaCl solution, indicating the potential importance of CNT dispersion for the toxicological studies. [164]
Several types of covalently functionalized MWCNTs: COOH-MWCNTs, sw-NH2- MWCNTs, NH2-MWCNTs, PEG-MWCNTs and PEI-MWCNTs Oropharyngeal aspiration 2 mg/kg body weight PBS with or without 0.6 mg/mL BSA
and 0.01 mg/mL DPPC
40 h or 21 days Male C57BL/6 mice aged 8 weeks Obvious lung fibrosis induced by cationic PEI-MWCNTs, whereas reduced pulmonary fibrosis observed in the group of MWCNT-COOH. [167]
Tau-MWCNTs and raw MWCNTs Intratracheal instillation 0.125, 0.25, 0.5 or 1 mg/kg body weight PBS 1, 7, 14 or 28 days Male CD-1 (ICR) mice of body weight 1822 g Less toxic induced by Tau-MWNTs than insoluble raw MWNTs [168]