Phyto-Fabrication Tribulus terrestris Mediated Iron Oxide Nanoparticles: A Promising Approach of Antioxidant and Anticancer Activities via in vitro and in silico Studies

Abstract

Background: Plant-mediated iron nanoparticles are increasingly utilized in biomedical and health applications due to their biocompatibility and nontoxicity. The therapeutic characteristics of these nanoparticles are extensively diverse.

Methods: In this study, iron nanoparticles synthesized from Tribulus terrestris were characterized using various techniques, including Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), UV-visible spectroscopy, vibrating sample magnetometry (VSM), and X-ray diffraction (XRD) analysis. Antioxidant properties were assessed using the hydrogen peroxide (H₂O₂) and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assays. Anti-inflammatory activity was evaluated through protein denaturation studies. Antimicrobial activity was tested against wound pathogens. The effects of anticancer and wound healing were investigated using HCT-116 (colon cancer) and MG-63 (osteosarcoma) cells. Molecular docking studies were performed to assess the binding affinity of Tribulus terrestris bioactive compounds with proteins involved in the Adenomatous polyposis coli (APC) pathway of colon cancer.

Results: The Tribulus terrestris-mediated Fe₃O₄ nanoparticles exhibited a peak at 290 nm using UV-visible spectroscopy. SEM and TEM analyses revealed that the nanoparticles were aggregated with an average size of 29 ± 0.24 nm. XRD analysis indicated a cubic crystalline structure. FTIR spectroscopy identified the biomolecules involved in the synthesis, and VSM confirmed a magnetic saturation of 14.75 emu/g. The antioxidant activity was demonstrated with DPPH (65.5%) and hydrogen peroxide (65.56%) assays at a dosage of 50 μg/mL, demonstrating a significant inhibition. The protein denaturation assay revealed a maximum inhibition of 54.57%. Lactobacillus had the strongest antibacterial activity at a concentration of 100 μg/mL, with an inhibitory zone of 35 mm. The anticancer assays showed IC₅₀ values of 25.95 μg/mL for colon cancer (HCT-116) and 35.36 μg/mL for osteosarcoma (MG-63), indicating significant cytotoxicity, particularly against colon cancer cells. The nanoparticles also demonstrated effective regulation of cell migration at 50 μg/mL. Molecular docking studies revealed strong binding affinities between Tribulus terrestris compounds and APC pathway proteins relevant to colon cancer.

Conclusion: This research underscores the potential of Tribulus terrestris-mediated iron nanoparticles as a sustainable and eco-friendly approach with significant antioxidant and anticancer properties, especially in combating colon cancer. The findings highlight their effectiveness in reducing oxidative stress, inhibiting cancer cell proliferation, and enhancing wound healing.

Keywords: Tribulus terrestris; antibacterial; anticancer; antioxidant; colon cancer; iron nanoparticles; molecular docking; osteosarcoma.