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

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 = ۶ per group). They received intraperitoneal injections of free eugenol, TiO₂ nanoparticles, or TiO₂@eugenol at low (۵۰ mg/kg) or high (۲۰۰ mg/kg) doses for ۱۴ days. Following the treatment period, serum concentrations of IL-۱β, IL-۶, and TNF-α were measured using ELISA; hepatic caspase-۳/۷ activity was assessed; and histological examinations of the liver, kidney, and spleen were performed. Gene expression of antioxidant markers (SOD۳, GR, GPx) in liver tissue was evaluated by qRT-PCR.Results: TiO₂ NPs significantly increased pro-inflammatory cytokines and hepatic caspase-۳/۷ 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.

کلیدواژه ها:

Titanium dioxide ، NPs ، Eugenol ، Inflammation ، Cytotoxicity