Nanotechnology applications are widely incorporated into the food sector. Due to their smaller size and increased volume-specific surface area, nanoparticles may possess unique properties compared to their bulk counterparts, making them useful for applications such as food coloring and plant protection products. Manufacturers bringing food-based nanotechnology applications to the European market must comply with the European Union's regulatory framework, including a nanospecific risk assessment [1,2]. Currently, hazard and risk assessment still heavily relies on conventional animal testing.  Use of new approach methodologies (NAMs) can fill certain data gaps and complement the available safety studies, thereby avoiding the need for conducting additional in vivo studies. Although many NAMs are available, experience in using them to support risk assessment is scarce and the majority of the NAMs have not yet been validated. The integration of NAMs for the physicochemical characterization and genotoxicity assessment of inorganic nanomaterials and materials containing a fraction of nanoparticles, applied in the food chain, is particularly promising as illustrated in case studies examining iron (hydr)oxides applied as food additive, and copper oxides applied as plant protection product and feed additive [3].

A detailed physicochemical characterization of the test materials is an essential first step in the risk assessment. It includes measuring the particle size distribution, surface charge, specific surface area, shape, solubility and dissolution rate, agglomeration state, chemical composition and crystal structure. Data obtained by electron microscopy based methods on the physicochemical characterization of iron (hydr)oxide and copper oxide NMs applied in the food chain will be presented.

Moreover, an overview will be provided of the NAMs that will be used to investigate the genotoxic potential of the characterized iron (hydr)oxides and copper oxides. These cover both the in vitro genotoxicity tests that are currently recommended by EFSA which are adapted for nanomaterials and materials containing a fraction of nanoparticles [4], as well as new innovative NAMs such as the transcriptomics-based biomarker, called GENOMARK. This NAM uses prediction models to classify analyzed materials as genotoxic or non-genotoxic based on gene expression data. In addition, a high-content method to simultaneously assess several genotoxicity parameters (e.g. ɣH2AX, PH3,…) with the Cytek-Amnis ImageStream technique will be developed.

This research is performed under the NAMS4NANO action via funding from the European Union through a grant of the European Food Safety Authority (agreement GP/EFSA/MESE/2022/01). This communication reflects only the author’s view and EFSA is not responsible for any use that may be made of the information it contains.

References

[1]          EFSA Scientific Committee et al., Guidance on technical requirements for regulated food and feed product applications to establish the presence of small particles including nanoparticles, EFS2 19, (2021).

[2]          EFSA Scientific Committee et al., Guidance on risk assessment of nanomaterials to be applied in the food and feed chain: human and animal health, EFS2 19, (2021).

[3]          REGULATION  (EU)  2015/  2283  OF  THE  EUROPEAN  PARLIAMENT  AND  OF  THE  COUNCIL  -  of  25  November  2015  -  on  novel  foods,  amending  Regulation  (EU)  No  1169/  2011  of  the  European  Parliament  and  of  the  Council  and  repealing  Regulation  (EC)  No  258/  97  of  the  European  Parliament  and  of  the  Council  and  Commission  Regulation  (EC)  No  1852/  2001, (n.d.).[4]          EFSA Scientific Committee, Scientific opinion on genotoxicity testing strategies applicable to food and feed safety assessment, EFS2 9, (2011).

[4]          EFSA Scientific Committee, Scientific opinion on genotoxicity testing strategies applicable to food and feed safety assessment, EFS2 9, (2011).