Nanomaterials in foods and the standardized static in vitro digestion method: contributing to the study of the potential toxic effects

Martins, Carla; Assunção, Ricardo; Gramacho, A.; Alvito, Paula; Silva, Maria João; Louro, H.

http://hdl.handle.net/10400.18/7118

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The use of nanoparticles (e.g. titanium dioxide) in commercial food products to modify some properties, such as brightness and whiteness, increased in the last years and is nowadays widespread. Despite the inhalation of nanoparticles is already a topic of concern, the potential adverse health effects due to ingestion still presents gaps of knowledge. In fact, gastrointestinal tract is the first interface between the body and the external environment and consequently could represent a target organ for compounds present in food that could exert toxic effects. The in vitro digestion models used to simulate the human digestion may contribute to fill these gaps. A standardized in vitro digestion model (IVD) was developed within the INFOGEST COST Action. This method considers a three-compartment model, simulating the digestion in the mouth, stomach and small intestine, applying standardized parameters such as pH, enzymatic activity and incubation periods. Within the scope of INGESTnano project (PTDC/SAU-PUB/29841/2017), and under physiological conditions, three TiO2 nanomaterials (NMs) were selected to setup a workflow to address the nanosafety regarding ingested NMs. The IVD was used to simulate the human gastrointestinal digestion of the NMs and the final product was used to test and ascertain NMs toxicity, using cell lines as intestinal models. The use of this standardized IVD model presented some challenges such as the high level of toxicity of the final digestion product for the bioassays. Several modifications to the initial protocol were investigated to overcome this issue. The results suggested that the addition of bile salts accounted for the majority of the observed toxicity. The applicability of the harmonized in vitro digestion method will be discussed in view of its potential use as a tool for addressing the toxicity of ingested NMs or other food contaminants, mimicking the physiological processes.