Durability and inflammogenic impact of carbon nanotubes compared with asbestos fibres

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Abstract

Background: It has been suggested that carbon nanotubes might conform to the fibre pathogenicity paradigm that explains the toxicities of asbestos and other fibres on a continuum based on length, aspect ratio and biopersistence. Some types of carbon nanotubes satisfy the first two aspects of the fibre paradigm but only recently has their biopersistence begun to be investigated. Biopersistence is complex and requires in vivo testing and analysis. However durability, the chemical mimicking of the process of fibre dissolution using in vitro treatment, is closely related to biopersistence and more readily determined. Here, we describe an experimental process to determine the durability of four types of carbon nanotubes in simulated biological fluid (Gambles solution), and their subsequent pathogenicity in vivo using a mouse model sensitive to inflammogenic effects of fibres. The in vitro and in vivo results were compared with well-characterised glass wool and asbestos fibre controls.Results: After incubation for up to 24 weeks in Gambles solution, our control fibres were recovered at percentages consistent with their known in vitro durabilities and/or in vivo persistence, and three out of the four types of carbon nanotubes tested (single-walled (CNTSW) and multi-walled (CNTTANG2, CNTSPIN)) showed no, or minimal, loss of mass or change in fibre length or morphology when examined by electron microscopy. However, the fourth type [multi-walled (CNTLONG1)] lost 30% of its original mass within the first three weeks of incubation, after which there was no further loss. Electron microscopy of CNTLONG1samples incubated for 10 weeks confirmed that the proportion of long fibres had decreased compared to samples briefly exposed to the Gambles solution. This loss of mass and fibre shortening was accompanied by a loss of pathogenicity when injected into the peritoneal cavities of C57Bl/6 mice compared to fibres incubated briefly. CNTSWdid not elicit an inflammogenic effect in the peritoneal cavity assay used here.Conclusions: These results support the view that carbon nanotubes are generally durable but may be subject to bio-modification in a sample-specific manner. They also suggest that pristine carbon nanotubes, either individually or in rope-like aggregates of sufficient length and aspect ratio, can induce asbestos-like responses in mice, but that the effect may be mitigated for certain types that are less durable in biological systems. Results indicate that durable carbon nanotubes that are either short or form tightly bundled aggregates with no isolated long fibres are less inflammogenic in fibre-specific assays. © 2011 Osmond-McLeod et al; licensee BioMed Central Ltd.

Figures

  • Figure 1 Paradigm for the role of length and biopersistence in the pathogenic effects of fibres. Schematic outlining the possible fates for fibres in vivo based on length and biopersistence (from Donaldson et al. 2006). Long, narrow (not depicted) and biopersistent fibres that deposit in the distal lung regions may cause inflammation, fibrosis and/or cancer.
  • Table 1 Fibres investigated in this study and rationale for their use
  • Figure 2 Durabilities of CNT samples and controls in simulated biological fluid. The weights of quadruplicate samples of glass wool fibre (X607), asbestos fibres (LFA, LFC) and CNT test samples (CNTSW, CNTSPIN, CNTLONG1 and CNTTANG2), recovered from Gambles solution following incubation for various times are expressed relative to the weight of sample recovered at 0 weeks.
  • Table 2 Weights of samples recovered from Gambles solution at 0 weeks and various incubation times to 24 weeks
  • Table 3 Average widths and lengths, and length distributions, of samples incubated in Gambles solution for 0 weeks or 10 weeks, as determined by TEM measurements
  • Figure 3 Effect of incubation in Gambles solution on fibre widths and lengths. Boxplots showing the distribution of fibre widths and lengths (nm) in samples that had been incubated in Gambles solution for 0 weeks or 10 weeks. The line in the box represents the median value of measurements from TEM images, and the edges of the box represent the lower and upper quartiles. The ends of the whiskers represent minimum and maximum values.
  • Figure 4 Appearance of fibres before and after incubation in Gambles solution. Representative SEM images of samples after 0 weeks or 10 weeks incubation in Gambles solution are shown at 5.0 K magnification in the two panels on the left. TEM images of equivalent samples, at indicated magnifications, are on the right.
  • Figure 5 Inflammatory responses to test samples injected into the peritoneal cavities of mice. Samples of vehicle only, X607, LFA, LFC, CNTSW and CNTLONG1, incubated for either 0 weeks or 10 weeks in Gambles solution, were injected into the peritoneal cavities of mice, and inflammatory responses were assessed at 24 h (panels on left) and 7 d (panels on right) post-injection. The inflammatory response was assayed by total (A) and differential (B) cell counts to identify infiltration of immune cells into the peritoneal cavity, total protein (C) to indicate increased permeability in the peritoneal cavity, IL-6 (D) as a measure of inflammatory cytokines, LDH (E) to indicate damage to cellular membranes, and the development of fibrotic plaques at 7 d (G). Note that the scales of the vertical axes vary. Differences between mice treated with the same fibre samples incubated for 0 weeks or 10 weeks were assessed by unpaired t-tests, with statistical significance set at p < 0.05 (denoted by an asterisk). Differences between mice treated with fibres that had been incubated for 0 weeks or 10 weeks compared to mice treated with vehicle only were assessed by one-way ANOVA with Tukey’s Multiple Comparison post-test, with statistical significance set at p < 0.05 (denoted by an unshaded triangle). For the differential cell counts, statistically significant differences between the numbers of macrophages (MAC) and polymorphonuclear leukocytes (PMNs) in treated mice compared to control mice are denoted by an unshaded triangle or a black square, respectively.

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CITATION STYLE

APA

Osmond-McLeod, M. J., Poland, C. A., Murphy, F., Waddington, L., Morris, H., Hawkins, S. C., … Donaldson, K. (2011). Durability and inflammogenic impact of carbon nanotubes compared with asbestos fibres. Particle and Fibre Toxicology, 8. https://doi.org/10.1186/1743-8977-8-15

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