Title: Protein expression profiles of intestinal epithelial co-cultures: effect of functionalised carbon nanotube exposure

Authors: Xianyin Lai; Bonnie L. Blazer-Yost; James W. Clack; Sharry L. Fears; Somenath Mitra; Susana Addo Ntim; Heather N. Ringham; Frank A. Witzmann

Addresses: Department of Cellular and Integrative Physiology, Indiana University School of Medicine, 1345 West 16th Street, Indianapolis IN 46202, USA ' Department of Cellular and Integrative Physiology, Indiana University School of Medicine, 1345 West 16th Street, Indianapolis IN 46202, USA; Department of Biology, Indiana University-Purdue University Indianapolis, 723 West Michigan Street, Indianapolis IN 46202, USA ' Department of Biology, Indiana University-Purdue University Columbus, 4601 Central Avenue, Columbus IN 47203, USA ' Sensient Bio-Ingredients, 5600 West Raymond Street, Indianapolis IN 46241, USA ' Department of Chemistry and Environmental Science, New Jersey Institute of Technology, Newark NJ 07102, USA ' Department of Chemistry and Environmental Science, New Jersey Institute of Technology, Newark NJ 07102, USA ' Department of Cellular and Integrative Physiology, Indiana University School of Medicine, 1345 West 16th Street, Indianapolis IN 46202, USA ' Department of Cellular and Integrative Physiology, Indiana University School of Medicine, 1345 West 16th Street, Indianapolis IN 46202, USA

Abstract: To assess the biological effects of low level, water dispersible, functionalised carbon nanotube (f-CNT) exposure in an in vitro model simulating the digestive tract, cellular protein expression was quantified and compared using label-free quantitative mass spectrometry (LFQMS). Co-cultured cells were exposed to well-characterised SWCNT-COOH, MWCNT-COOH, and MWCNT-PVP. The relative expression of 2,282 unique proteins was compared across the dose groups. 428 proteins were found to be differentially expressed. At the high dose, the extent of differential protein expression was CNT-specific and directly related to CNT colloidal stability. Cells responded to low level MWCNT-PVP exposure with three-fold greater differential expression. Bioinformatic analysis indicated significant and f-CNT-specific effects on relevant molecular and cellular functions and canonical pathways, with little overlap across f-CNT type and in the absence of overt toxicity.

Keywords: carbon nanotubes; CNTs; colon; in vitro; intestinal cells; label-free quantitation; mass spectrometry; poly(4-vinylpyrrolidone); proteomics; protein expression profiles; nanotechnology; functionalised CNT exposure; digestive tract; simulation.

DOI: 10.1504/IJBNN.2013.054508

International Journal of Biomedical Nanoscience and Nanotechnology, 2013 Vol.3 No.1/2, pp.127 - 162

Published online: 12 Jul 2014 *