Spectroscopic characterization of protein-wrapped single-wall carbon nanotubes and quantification of their cellular uptake in multiple cell generations

Abstract

We study the spectral characteristics of bovine serum albumin (BSA) protein conjugated single-wall carbon nanotubes (SWNTs), and quantify their uptake by macrophages. The binding of BSA onto the SWNT surface is found to change the protein structure and to increase the doping of the nanotubes. The G-band Raman intensity follows a well-defined power law for SWNT concentrations of up to 33 microg ml(-1) in aqueous solutions. Subsequently, in vitro experiments demonstrate that incubation of BSA-SWNT complexes with macrophages affects neither the cellular growth nor the cellular viability over multiple cell generations. Using wide spot Raman spectroscopy as a fast, non-destructive method for statistical quantification, we observe that macrophages effectively uptake BSA-SWNT complexes, with the average number of nanotubes internalized per cell remaining relatively constant over consecutive cell generations. The number of internalized SWNTs is found to be approximately 30 10(6) SWNTs/cell for a 60 mm(-2) seeding density and approximately 100 x 10(6) SWNTs/cell for a 200 mm(-2) seeding density. Our results show that BSA-functionalized SWNTs are an efficient molecular transport system with low cytotoxicity maintained over multiple cell generations.

High resolution and dynamic imaging of biopersistence and bioreactivity of extra and intracellular MWNTs exposed to microglial cells

“…The AO MWNT loadings reported in this work are lower than the ~0.6 vol% loading of protein-wrapped SWNTs within macrophage cells found by Bertulli et al . [24] using Raman spectroscopy for uptake quantification. Increased uptake of protein-SWNTs will be related to a range of factors including the larger population of individualized SWNT in these dispersions.…”

“…Furthermore, there are relatively few studies which quantify the amount of unlabelled MWNTs that come into contact with, or are internalised by, cells and which correlate this information to cell toxicity. Raman spectroscopy has been used to determine SWNT concentrations in washed macrophage cell samples [24] as well as in fibroblast cell lysates [25]; however, resonance effects make direct quantification challenging.…”

Long-term effects of carbon containing engineered nanomaterials and asbestos in the lung: one year postexposure comparisons

“…Assessments of SWCNTs retained in the lungs 1 yr postexposure were performed using Raman microscopy. This protocol has been successfully employed for the detection of SWCNTs in tissues and cells due to the presence of a characteristic tangential-mode G-band in their Raman spectra (4,7). The Raman spectra were measured using a Horiba Jobin-Yvon spectrometer connected to a CCD camera and a confocal microscope with an ϫ80 objective.…”

The developmental toxicity, bioaccumulation and distribution of oxidized single walled carbon nanotubes in Artemia salina

“…Raman spectroscopy has been verified to be a sensitive, non-destructive and reliable method for the quantification of CNTs because it has a low LOD in biological tissues (0.1-1 mg kg −1 ). [38][39][40] Previously, the technique has been well performed to quantify MWCNTs in Saccharomyces cerevisiae 26 and A. salina. 24 In the study, Raman spectroscopy was used to measure the O-SWCNT content based on the dry weight of A. salina.…”