Comparative evaluation of cytotoxicity and inflammatory responses induced by free and eugenol-loaded titanium dioxide nanoparticles following intraperitoneal injection in mouse

Articles in Press

Document Type : Research Paper

Authors

1 Department of Genetics, Faculty of Basic Sciences, Shahrekord University, Shahrekord, Iran

2 Department of Pathobiology, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran

3 Department of Pathobiology, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran

10.22038/nmj.2025.90454.2286

Abstract

Objective(s): Titanium dioxide nanoparticles (TiO₂ NPs), which are widely used in food and consumer products, have been associated with oxidative stress and inflammatory toxicity. Eugenol, a naturally occurring phenolic compound with well-established anti-inflammatory and antioxidant properties, may exert protective effects when delivered through nanocarriers.
Materials and Methods: TiO₂ nanoparticles were synthesized via a co-precipitation method and subsequently functionalized with eugenol (TiO₂@eugenol). FTIR, XRD, DLS, zeta potential analysis, FE-SEM, and TEM were used to characterize the nanoparticles. Thirty-six BALB/cJ mice were randomly assigned to six groups (n = 6 per group). They received intraperitoneal injections of free eugenol, TiO₂ nanoparticles, or TiO₂@eugenol at low (50 mg/kg) or high (200 mg/kg) doses for 14 days. Following the treatment period, serum concentrations of IL-1β, IL-6, and TNF-α were measured using ELISA; hepatic caspase-3/7 activity was assessed; and histological examinations of the liver, kidney, and spleen were performed. Gene expression of antioxidant markers (SOD3, GR, GPx) in liver tissue was evaluated by qRT-PCR.
Results: TiO₂ NPs significantly increased pro-inflammatory cytokines and hepatic caspase-3/7 activity. They also induced necrosis and inflammatory alterations in the liver, kidney, and spleen. In contrast, TiO₂@eugenol markedly suppressed cytokine release and apoptotic activity while preserving tissue architecture. qRT-PCR analysis showed that TiO₂ NPs downregulated antioxidant-related genes, whereas TiO₂@eugenol significantly upregulated their expression, indicating improved redox homeostasis.
Conclusion: Eugenol functionalization improved the biocompatibility profile of TiO₂ NPs and provided substantial protection against TiO₂-induced toxicity by attenuating inflammation, apoptosis, and oxidative stress while restoring antioxidant defenses. These findings highlight the therapeutic potential of eugenol-loaded TiO₂ nanoparticles and support further investigation in extended exposure models and disease-specific applications.

Keywords

Main Subjects

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Available Online from 24 December 2025

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