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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
Affiliation:
rajap@rpi.edu, Rensselaer Polytechnic Institute, Chemical and Biological Engineering, CII 9015 - CIE, 110 8th Street, Troy, NY, 12180, United States
Jennifer Connolley
Affiliation:
connoj@rpi.edu, Rensselaer Polytechnic Institute, Biomedical Engineering, Troy, NY, 12180, United States
Pulickel M. Ajayan
Affiliation:
ajayan@rpi.edu, Rensselaer Polytechnic Institute, Materials Science and Engineering, Troy, NY, 12180, United States
Omkaram Nalamasu
Affiliation:
nalamasu@rpi.edu, Rensselaer Polytechnic Institute, Center for Integrated Electronics, Troy, NY, 12180, United States
Deanna M. Thompson
Affiliation:
thompd4@rpi.ed, Rensselaer Polytechnic Institute, Biomedical Engineering, Troy, NY, 12180, United States
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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.

Type
Research Article
Copyright
Copyright © Materials Research Society 2007

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References

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