Fullerenol cytotoxicity in kidney cells is associated with cytoskeleton disruption, autophagic vacuole accumulation, and mitochondrial dysfunction

Journal Article · Mon Nov 01 00:00:00 EDT 2010 · Toxicology and Applied Pharmacology

DOI:https://doi.org/10.1016/j.taap.2010.08.008· OSTI ID:21460230

Johnson-Lyles, Denise N ^[1]; Peifley, Kimberly; Lockett, Stephen ^[2]; Neun, Barry W; Hansen, Matthew; Clogston, Jeffrey ^[1]; McNeil, Scott E ^[1]

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1. Nanotechnology Characterization Lab (NCL), Advanced Technology Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, MD 21702 (United States)
2. Optical Microscopy Analysis Lab (OMAL), Advanced Technology Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, MD 21702 (United States)

Water soluble fullerenes, such as the hydroxylated fullerene, fullerenol (C{sub 60}OH{sub x}), are currently under development for diagnostic and therapeutic biomedical applications in the field of nanotechnology. These molecules have been shown to undergo urinary clearance, yet there is limited data available on their renal biocompatibility. Here we examine the biological responses of renal proximal tubule cells (LLC-PK1) exposed to fullerenol. Fullerenol was found to be cytotoxic in the millimolar range, with viability assessed by the sulforhodamine B and trypan blue assays. Fullerenol-induced cell death was associated with cytoskeleton disruption and autophagic vacuole accumulation. Interaction with the autophagy pathway was evaluated in vitro by Lysotracker Red dye uptake, LC3-II marker expression and TEM. Fullerenol treatment also resulted in coincident loss of cellular mitochondrial membrane potential and ATP depletion, as measured by the Mitotracker Red dye and the luciferin-luciferase assays, respectively. Fullerenol-induced ATP depletion and loss of mitochondrial potential were partially ameliorated by co-treatment with the autophagy inhibitor, 3-methyladenine. In vitro fullerenol treatment did not result in appreciable oxidative stress, as measured by lipid peroxide and glutathione content. Based on these data, it is hypothesized that cytoskeleton disruption may be an initiating event in fullerenol cytotoxicity, leading to subsequent autophagy dysfunction and loss of mitochondrial capacity. As nanoparticle-induced cytoskeleton disruption, autophagic vacuole accumulation and mitochondrial dysfunction are commonly reported in the literature, the proposed mechanism may be relevant for a variety of nanomaterials.

• OSTI ID: 21460230
• Journal Information: Toxicology and Applied Pharmacology, Vol. 248, Issue 3; Other Information: DOI: 10.1016/j.taap.2010.08.008; PII: S0041-008X(10)00281-4; Copyright (c) 2010 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; ISSN 0041-008X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
APOPTOSIS
ATP
FULLERENES
GLUTATHIONE
LUCIFERASE
LUCIFERIN
MICROTUBULES
MITOCHONDRIA
NANOSTRUCTURES
TOXICITY
TUBULES
ALBUMINS
BODY
CARBON
CELL CONSTITUENTS
DRUGS
ELEMENTS
ENZYMES
KIDNEYS
NONMETALS
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANS
OXIDASES
OXIDOREDUCTASES
PEPTIDES
POLYPEPTIDES
PROTEINS
RADIOPROTECTIVE SUBSTANCES
RESPONSE MODIFYING FACTORS