From Synthesis to Cellular Response: Physicochemical Characterization and in Vitro Toxicological Evaluations of Graphene Oxide- and ZnO-Based Nanocomposites

Abstract

Nanomaterials are increasingly used in food packaging due to their antimicrobial and barrier-enhancing properties; however, concerns persist about their potential biological impacts on human health. This study comparatively evaluated the cytotoxic and cytogenetic responses induced by zinc oxide (ZnO), graphene oxide (GO), reduced graphene oxide (rGO), and their nanocomposites (ZnO/GO and ZnO/rGO), which were synthesized in our laboratory, using complementary human cell models. The materials were synthesized and characterized by dynamic light scattering, Fourier transform infrared spectroscopy, and scanning electron microscopy with energy-dispersive X-ray analysis to confirm their composition and morphology. Cytotoxicity was assessed by the MTT assay in human colorectal adenocarcinoma (DLD-1) and normal colon fibroblast (CCD-18Co) cell lines, while the mitotic index (MI) was evaluated in cultured human peripheral lymphocytes. The results of this study revealed that ZnO, GO, rGO, and their corresponding nanocomposites caused no significant acute cytotoxic effects in both cancerous and normal human cell lines at the tested concentrations (0.25-1200 µg/mL) following 24 h exposure. However, the consistent, concentration-dependent decrease in mitotic index following 24- and 48-hour treatment (10-150 µg/mL) suggests that these nanomaterials may influence cell cycle progression, highlighting potential cytogenetic effects that warrant attention. Although such effects did not translate into immediate cell death, they underscore the need for careful interpretation when considering these materials for food-contact uses. Therefore, further investigations involving long-term exposure, additional genotoxic markers, and in vitro and in vivo models are recommended to more fully establish their safety profile.

Keywords

• Zinc oxide nanoparticles
• Graphene oxide
• Nanocomposites
• MTT assay
• Mitotic index

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