Hostname: page-component-586b7cd67f-2plfb Total loading time: 0 Render date: 2024-11-27T12:21:13.122Z Has data issue: false hasContentIssue false
  • English
  • Français

DAPI Method: A Novel Assay to Evaluate the In vitro Impact of Nanomaterials on Mammalian Cells

Published online by Cambridge University Press:  01 February 2011

Pavan M. V. Raja ,
Jennifer Connolley ,
Pulickel M. Ajayan ,
Omkaram Nalamasu  and
Deanna M. Thompson
Pavan M. V. Raja
Affiliation:
[email protected], Rensselaer Polytechnic Institute, Chemical and Biological Engineering, CII 9015 - CIE, 110 8th Street, Troy, NY, 12180, United States
Jennifer Connolley
Affiliation:
[email protected], Rensselaer Polytechnic Institute, Biomedical Engineering, Troy, NY, 12180, United States
Pulickel M. Ajayan
Affiliation:
[email protected], Rensselaer Polytechnic Institute, Materials Science and Engineering, Troy, NY, 12180, United States
Omkaram Nalamasu
Affiliation:
[email protected], Rensselaer Polytechnic Institute, Center for Integrated Electronics, Troy, NY, 12180, United States
Deanna M. Thompson
Affiliation:
[email protected], Rensselaer Polytechnic Institute, Biomedical Engineering, Troy, NY, 12180, United States
Get access

Abstract

The increasing importance of nanomaterial-related applications has given rise to concerns pertaining to their impact on human health. In vitro mammalian cell-based assays can provide a quick and simple estimate of the possible adverse effects of the nanomaterials. However, recent studies have questioned the efficacy of traditional assays such as the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, in evaluating cell-nanomaterial interactions, implying the need for alternate methods. We applied image analysis to enumerate the DAPI (2-[4-(Aminomethyl) phenyl]-1H-indole-6-carboximidamide, dihydrochloride) – stained cellular nuclei. Image analysis, being non-destructive, capable of automation, and applicable over a wide range of cell seeding densities, offers several advantages compared to older methods like the MTT assay and hemocytometry. Using image analysis, the impact of singlewalled carbon nanotubes (SWNT) on rat aortic smooth muscle cell (SMC) growth kinetics, were examined. Despite the carbon nanomaterial presence, the fluorescent signal from the nuclei was not noticeably impacted over the SWNT range examined (0.00-0.10 mg/ml). We anticipate that this method can also be applied to evaluate the biological impact of other nanomaterials.

Keywords

nanoscalebiologicalnanostructure
Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 951: Symposium E – Nanofunctional Materials, Nanostructures and Novel Devices for Biological and Chemical Detection , 2006 , 0951-E06-38
Copyright
Copyright © Materials Research Society 2007

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

1. Shvedova, A. A., et al, Exposure to carbon nanotube material: assessment of nanotube cytotoxicity using human keratinocyte Cells, J. Tox. & Env. Health. 66, pp.1909–1926 (2003).Google Scholar
2. Cui, D. et al, Effect of single wall carbon nanotubes on human HEK293 cells, Tox. Lett. 155, pp.73–85 (2005).Google Scholar
3. Nimmagadda, A. et al, Chemical modification of SWNT alters in vitro cell-SWNT interactions, J. Biomed. Mat. Res. 76, pp.614–625 (2006).Google Scholar
4. Sayes, C. M., et al, Functionalization density dependence of single-walled carbon nanotubes cytotoxicity in vitro, Tox. Lett. 161, pp.135–142 (2006).Google Scholar
5. Lam, C. W. et al, Pulmonary toxicity of single-wall carbon nanotubes in mice 7 and 90 dfays after intratracheal instillation, Tox. Sci. 77, pp.126–134 (2004).Google Scholar
6. Warheit, D. B. et al, Comparative pulmonary toxicity assessment of single-wall carbon nanotubes in rats, Tox. Sci. 77, pp.117–125 (2004).Google Scholar
7. Swartz, E. A. et al, Exposure of human vascular endothelial cells to sustained hydrostatic pressure stimulates proliferation: involvement of the alpha-v integrins, Circ. Res. 84, pp.315–322 (1999).Google Scholar