کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5861564 1133761 2015 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Non-specific interaction of carbon nanotubes with the resazurin assay reagent: Impact on in vitro assessment of nanoparticle cytotoxicity
ترجمه فارسی عنوان
تعامل غیر اختصاصی نانولولههای کربن با معرف معرفی رزازورین: تأثیر در ارزیابی درون سلولی سمیت سلولی نانوذرات
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم محیط زیست بهداشت، سم شناسی و جهش زایی
چکیده انگلیسی


- CNTs interfere with fluorescence-based resazurin cell viability assay.
- Physical quench can be circumvented by clarification of supernatants by centrifugation.
- Polar, oxidized CNTs but not pristine CNTs chemically quench resorufin fluorescence.
- Chemical interference can be easily quantified for elimination of the bias.

In vitro cytotoxicity assays are essential tools in the screening of engineered nanomaterials (NM) for cellular toxicity. The resazurin live cell assay is widely used because it is non-destructive and is well suited for high-throughput platforms. However, NMs, in particular carbon nanotubes (CNT) can interfere in assays through quenching of transmitted light or fluorescence. We show that using the resazurin assay with time-point reading of clarified supernatants resolves this problem. Human lung epithelial (A549) and murine macrophage (J774A.1) cell lines were exposed to NMs in 96-well plates in 200 μL of media/well. After 24 h incubation, 100 μL of supernatant was removed, replaced with resazurin reagent in culture media and aliquots at 10 min and 120 min were transferred to black-wall 96-well plates. The plates were quick-spun to sediment the residual CNTs and fluorescence was top-read (λEx = 540 nm, λEm = 600 nm). The procedure was validated for CNTs as well as silica nanoparticles (SiNP). There was no indication of reduction of resazurin by the CNTs. Stability of resorufin, the fluorescent product of the resazurin reduction was then assessed. We found that polar CNTs could decrease the fluorescence signal for resorufin, possibly through oxidation to resazurin or hyper-reduction to hydroxyresorufin. This effect can be easily quantified for elimination of the bias. We recommend that careful consideration must be given to fluorimetric/colorimetric in vitro toxicological assessments of optically/chemically active NMs in order to relieve any potential artifacts due to the NMs themselves.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Toxicology in Vitro - Volume 29, Issue 1, February 2015, Pages 142-147
نویسندگان
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