Toxic effects of a modified montmorillonite clay on the human intestinal cell line Caco-2

J Appl Toxicol. 2014 Jun;34(6):714-25. doi: 10.1002/jat.2945. Epub 2013 Oct 9.


The incorporation of the natural mineral clay montmorillonite into polymeric systems enhances their barrier properties as well as their thermal and mechanical resistance, making them suitable for a wide range of industrial applications, e.g., in the food industry. Considering humans could easily be exposed to these clays due to migration into food, toxicological and health effects of clay exposure should be studied. In the present work, the cytotoxic effects induced by two different clays (the unmodified clay Cloisite(®) Na(+) , and the organically modified Cloisite(®) 30B) on Caco-2 cells were studied after 24 and 48 h of exposure. The basal cytotoxicity endpoints assessed were total protein content, neutral red uptake and a tetrazolium salt reduction. Our results showed that only Cloisite(®) 30B induced toxic effects. Therefore, the effects of subcytotoxic concentrations of this clay on the generation of intracellular reactive oxygen species, glutathione content and DNA damage (comet assay) were investigated. Results indicate that oxidative stress may be implicated in the toxicity induced by Closite(®) 30B, in regards of the increases in intracellular reactive oxygen species production and glutathione content at the highest concentration assayed, while no damage was observed in DNA. The most remarkable morphological alterations observed were dilated cisternae edge in the Golgi apparatus and nucleolar segregation, suggesting impairment in the secretory functions, which could be related to inhibition in the synthesis of proteins.

Keywords: Cloisite®; cell culture; clays; cytotoxicity; morphology; oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aluminum Silicates / toxicity*
  • Bentonite / toxicity*
  • Caco-2 Cells
  • Cell Nucleolus / metabolism
  • Cell Nucleolus / ultrastructure
  • Cell Shape / drug effects
  • Clay
  • Colon / drug effects*
  • Colon / metabolism
  • Colon / ultrastructure
  • DNA Damage
  • Dose-Response Relationship, Drug
  • Glutathione / metabolism
  • Golgi Apparatus / drug effects
  • Golgi Apparatus / metabolism
  • Golgi Apparatus / ultrastructure
  • Humans
  • Oxidative Stress / drug effects
  • Reactive Oxygen Species / metabolism
  • Time Factors


  • Aluminum Silicates
  • Reactive Oxygen Species
  • Bentonite
  • Glutathione
  • Clay