In vitro toxicity of nanoparticles in BRL 3A rat liver cells
Introduction
Nanotechnology involves the creation and manipulation of materials at nanoscale levels to create products that exhibit novel properties. Recently, nanomaterials such as nanotubes, nanowires, fullerene derivatives (buckyballs) and quantum dots have received enormous attention to create new types of analytical tools for biotechnology and life sciences (Bruchez et al., 1998, Taton et al., 2000, Cui et al., 2001). Nanomaterials, which range in size from 1 to 100 nm, have been used to create unique devices at the nanoscale level possessing novel physical and chemical functional properties (Colvin, 2003, Oberdörster, 2004). Although nanomaterials are currently being widely used in modern technology, there is a serious lack of information concerning the human health and environmental implications of manufactured nanomaterials. The major toxicological concern is the fact that some of the manufactured nanomaterials are redox active (Colvin, 2003), and some particles transport across cell membranes and especially into mitochondria (Foley et al., 2002). One of the few relevant studies was with single-wall carbon nanotubes in mice (Lam et al., 2004). Lam et al. (2004) demonstrated that carbon nanotube products induced dose-dependent epithelioid granulomas in mice and, in some cases, interstitial inflammation in the animals of the 7-day post-exposure groups. The recent study by Oberdörster (2004) indicated that nanomaterials (Fullerences C60) induced oxidative stress in a fish model. Although limited studies have been conducted on the toxicity of nanoparticles, there are no reports on the use of in vitro models to evaluate potential toxicity screening of nanomaterials. The BRL 3A immortal rat liver cell line was selected in the present study as a convenient in vitro model to assess nanocellular toxicity. This cell line has been well characterized for its relevance to toxicity models (Boess et al., 2003). In vivo exposure to nanoparticles is likely to have potential impact on the liver since exposure to these particles is likely to occur through ingestion and clearance by the liver (Jani et al., 1990). The toxicity end points (MTT, LDH, ROS and GSH) that were selected in the current study represent vital biological functions of the mammalian system as well as provide a general sense of toxicity in a relatively short time. The results described in this paper provide a range of doses that were toxic to these cultured cells and data pointing to a general mechanism of nanoparticle toxicity. Since little information is available on nanomaterial toxicity, simple in vitro toxicity models and general toxicity end points are likely to assist in mechanistic events after exposure and subsequent toxicity risk assessment of nanomaterials.
Section snippets
Chemicals
The test materials silver (Ag; 15, 100 nm), molybdenum (MoO3; 30, 150 nm), aluminum (Al; 30, 103 nm), iron oxide (Fe3O4; 30, 47 nm), manganese oxide (MnO2; 1–2 μm), and tungsten (W; 27 μm) were received from Air Force Research Laboratory, Brooks AFB, TX. Cadmium oxide (CdO-1000 nm) and titanium oxide (TiO2-40 nm) were purchased from Fluka Chemicals and Altair, Nanomaterials Inc., respectively. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), β-nicotinamide-adenine
Results
The results demonstrated that exposure to Ag nanoparticles for 24 h resulted in concentration-dependent increase in LDH leakage and exhibited a significant (p < 0.05) cytotoxicity at 10–50 μg/ml (Fig. 1B). It was noted that there is a statistically significant difference between different silver particle sizes of 100 and 15 nm, where the 100 nm particles showed higher toxicity at 25 and 50 μg/ml. The results for LDH leakage for MoO3, Al, Fe3O4, MnO2, W, nanoparticles exposure did not produce
Discussion
The purpose of this investigation was to evaluate potential toxicity and the general mechanism involved in nanoparticle toxicity. To date there are very few studies directly or indirectly investigating the toxic effects of nanomaterials and no clear guidelines are presently available to quantify these effects. Recently, Lam et al. (2004) reported that nanotubes induced lung tissue damage in mice resulting in granulomas. Another report by Warheit et al. (2004) investigated acute lung toxicity
Acknowledgements
This work was supported by the Air Force Office of Scientific Research (AFOSR) Project (JON# 2312A211) and performed in conjunction with U.S. Air Force Contract F41624-96-C-9010 (ManTech/Geo-Centers Joint Venture). We are thankful to our Division Chief Col Riddle for his strong support and encouragement. The authors are thankful to Dr. Marie Claude-Hofmann, University of Dayton for helping fluorescence microscopic work.
References (18)
- et al.
Cellular localisation of a water-soluble fullerene derivative
Biochemical and Biophysical Research Communications
(2002) - et al.
In vitro assessment of high energy chemicals in rat hepatocytes
The Science of the Total Environment
(2001) - et al.
Pulmonary response of rats exposed to titanium dioxide (TiO2) by inhalation for two years
Toxicology and Applied Pharmacology
(1985) Glutathione and its role in cellular functions
Free Radical Biology and Medicine
(1999)- et al.
Quantitating cellular oxidative stress by dicholorofluorescein assay using microplate reader
Free Radical Biology and Medicine
(1999) - et al.
Relationships and cytotoxicity in isolated rat hepatocytes
Archives of Biochemistry and Biophysics
(1990) - et al.
Gene expression in two hepatic cell lines, cultured primary hepatocytes, and liver slices compared to the in vivo liver gene expression in rats: possible implications for toxicogenomics use of in vitro systems
Toxicological Sciences
(2003) - et al.
Semiconductor nanocrystals as fluorescent biological labels
Science
(1998) - et al.
Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing
Cancer Research
(1987)
Cited by (1852)
Design rules applied to silver nanoparticles synthesis: A practical example of machine learning application.
2024, Computational and Structural Biotechnology JournalIron oxide nanoparticles for inflammatory bowel disease: Recent advances in diagnosis and targeted drug therapy
2024, Applied Surface Science AdvancesBlue light photobiomodulation induced apoptosis by increasing ROS level and regulating SOCS3 and PTEN/PI3K/AKT pathway in osteosarcoma cells
2023, Journal of Photochemistry and Photobiology B: BiologyGreen nanomaterials: Synthesis and applications in wastewater treatment
2023, Inorganic Chemistry CommunicationsTransactivator of transcription peptide conjugated copper oxide nanoparticles: A nano-warrior against breast cancer - Insights from biosynthesis, characterization, and cellular studies
2023, Journal of Drug Delivery Science and Technology