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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

aspiration

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.

[132]

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 18∼22 g

Less toxic induced by Tau-MWNTs than insoluble raw MWNTs

[168]