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Toxicology and Cellular Effect of Manufactured Nanomaterials

a nanomaterial and toxicology technology, applied in the field of carbon nanomaterials, can solve the problems of limited aerosol release of mwcnos and mwcnts during manufacturing, inability to fully understand the molecular mechanisms of cytotoxicity of this class of nanomaterials, and the risk associated with exposure, so as to achieve the effect of reducing toxic effects and eliminating nanotoxicities

Active Publication Date: 2012-12-13
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

The patent text describes a method for measuring the toxic effects of nanomaterials on cells and tissues. The method involves using biomarkers that are changed in response to nanoparticle exposure. These biomarkers can be used to evaluate the cytotoxicity of nanomaterials and to develop instruments and methods for measuring exposure to particles. The method can also be used to select and develop protective coatings for nanomaterials. The gene expression patterns of the biomarkers can be used to predict the effects of nanomaterials on cells and to develop targeting mechanisms for delivering nanomaterials to cancer cells for therapeutic purposes. The invention provides a reliable and valid method for measuring the toxic effects of nanomaterials and differentiating them from other toxic substances.

Problems solved by technology

Although both fullerene and carbon nanotubes have been demonstrated to accumulate to cytotoxic levels within organs of various animal models and cell types, and carbon nanomaterials have been exploited for cancer therapies, the molecular and cellular mechanisms for cytotoxicity of this class of nanomaterial are not yet fully apparent.
Fortunately, aerosol release of the MWCNOs and MWCNTs during manufacturing is limited.
However, because of the increase in use, the risk associated with exposure and the molecular mechanisms of any cytotoxicity need to be well understood.

Method used

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  • Toxicology and Cellular Effect of Manufactured Nanomaterials
  • Toxicology and Cellular Effect of Manufactured Nanomaterials
  • Toxicology and Cellular Effect of Manufactured Nanomaterials

Examples

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example 1

Materials and Methods of Measuring Toxicology of MWCNOs and MWCNTs

[0163]The carbon MWCNOs used in this study were produced by using a modified direct-current electric-arc discharge method based on the methods described by Sano, N., Wang, H., Chhowalla, M., Alexandrou, I. & Amaratunga, G. A. J. Nanotechnology: Synthesis of carbon ‘onions’ in water. Nature (London) 414, 506-507 (2001), hereby incorporated by reference. (See FIG. 1 and infra). The multi-walled carbon nanotubes (MWCNTs) were synthesized by using a chemical vapor deposition (CVD) method as described in Andrews, R., Jacques, D., Qian, D. & Rantell, T. Multiwall carbon nanotubes: synthesis and application. Acc Chem Res 35, 1008-1017 (2002), hereby incorporated by reference. See FIG. 1. Cellomics-based High Content Image Analysis (HCA) has been used for phenotypical measurement of cell apoptosis, necrosis, cell numbers, proliferation, and cell cycle distribution. Apoptotic cells and necrotic cells were detected using DNA dy...

example 2

Evaluating Toxicity in Nanomaterial Exposed Cells

[0182]In this study, the goal was to assess if changes in gene expression in cells exposed to carbon-based nanomaterials shows a correlation to phenotypic observations. Reported here are two approaches to evaluate toxicity in nanomaterial exposed cells; (i) the measurement of phenotypic changes in large populations of cells by high content analysis and (ii) gene expression array analysis in exposed cells. Phenotypically, cells exposed to high concentrations of nanomaterials were observed to undergo apoptosis / necrosis with a concomitant reduction in proliferation indicative of an inflammation response. We found that carbon nanomaterials, generated mRNA level changes in exposed skin fibroblasts, including changes in mRNA levels from genes involved in metabolism, apoptosis, cell cycle, stress response, cellular transport, and inflammatory response. Of interest was our observation that many of the genes that increased in expression in nan...

example 3

Evaluating Apoptosis and Necrosis in Nanomaterial Exposed Cells

[0193]Cytotoxicity was evaluated by staining live cells for 30 minutes with YO-PRO1 (Invitrogen, Molecular Probes), propidium iodide (PI, Sigma) and Hoechst. Live cells are impermeable to YO-PRO1 and PI, both of which are intercalating DNA dyes. Apoptotic cells are permeable to YO-PRO1, whilst PI only stains necrotic cells. Stained culture plates were analyzed using the KSR and images were acquired at each appropriate fluorescence channel for Hoechst, YO-PRO1, and PI. The image analysis software establishes average and total intensity for each nucleus in all channels. In the experiments pictured in FIG. 2, wells were either exposed to the indicated concentration of nanomaterials or the same volume of ethanol solvent as a control, both kept less than 1% of the total volume. Ten replicates were done for each condition with intensities for both YO-PRO1 and PI averaged by well, to obtain the bars in FIGS. 2B and 2C. Student ...

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Abstract

The increasing use of nanotechnology in consumer products and medical applications underlies the importance of understanding its potential toxic effects to people and the environment. Herein are described methods and assays to predict and evaluate the cellular effects of nanomaterial exposure. Exposing cells to nanomaterials at cytotoxic doses induces cell cycle arrest and increases apoptosis / necrosis, activates genes involved in cellular transport, metabolism, cell cycle regulation, and stress response. Certain nanomaterials induce genes indicative of a strong immune and inflammatory response within skin fibroblasts. Furthermore, the described multiwall carbon nanoonions (MWCNOs) can be used as a therapeutic in the treatment of cancer due to its cytotoxicity.

Description

CROSS-REFERENCE TO RELATED APPLICATION[0001]This application relates to and claims priority to International Application PCT / US06 / 60369 filed on Oct. 30, 2006, which claims priority to U.S. Provisional Patent Application, 60 / 731,557, filed on Oct. 28, 2005, and U.S. Provisional Patent Application, 60 / 792,557, filed on Apr. 12, 2006, all of which are herein incorporated by reference in their entirety for all purposes.STATEMENT OF GOVERNMENTAL SUPPORT[0002]This invention was made during work supported by NIH Grant R1CA95393-01, DOD BCRP BC045345 Grant, DARPA, UCSF Prostate Cancer SPORE award (NIH Grant P50 CA89520), by a DOE LDRD grant, and by NIH P50 Grant CA112970. This work was performed under the auspices of the U.S. Department of Energy, at the University of California / Lawrence Livermore National Laboratory under Contract No. W-7405-Eng-48, and University of California / Lawrence Berkeley National Laboratory, under Contract No. DE-AC03-76SF00098, now Contract No. DE-AC05-CH11231. T...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K49/00A61K9/14C40B30/00C12Q1/68C12Q1/02
CPCA61K47/48584A61K47/48869B82Y5/00C12Q2600/142C12Q2600/158G01N33/5014C12Q2600/106C12Q1/6883A61K47/6855A61K47/6925
Inventor CHEN, FANQING
Owner RGT UNIV OF CALIFORNIA
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