Evaluation of the toxicity and anti-angiogenic effects of beta-cyclodextrin complex loaded with thymol in melanoma skin cancer cells (A2058)

Background and Aim: In drug delivery, the formation of cyclodextrin complexes is a widely used strategy to overcome biopharmaceutical challenges associated with various active pharmaceutical ingredients, such as poor water solubility, initial instability, and low bioavailability. Given the anti-cancer properties of thymol, this study aimed to evaluate the effects of thymol-loaded nanoparticles in a beta-cyclodextrin complex and to assess their anti-cancer effects in melanoma skin cancer cells (A2058) by examining the expression of the VEGF and VEGFR genes. Methods: The synthesized nanoparticles were characterized using various methods, including zeta potential analysis, ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). To determine the cytotoxicity of the synthesized nanocomplex, the MTT assay was performed with β-CD-Thymol nanoparticle concentrations ranging from 62.5 to 500 μg/ml. The Chorioallantoic Membrane (CAM) assay was conducted to evaluate the cytotoxicity of the synthesized β-CD-Thymol nanoparticles in the context of angiogenesis. Finally, the expression levels of genes involved in angiogenesis (VEGF and VEGFR) were analyzed using real-time PCR. Results: Dynamic light scattering analysis revealed that the mean size of the nanoparticles was 116.20 nm, with a zeta potential of -21.64 mV. FTIR studies indicated the presence of several biologically active functional groups in the synthesized nanoparticles. Additionally, the anti-cancer effects and inhibitory role of the β-CD-Thymol complex were confirmed by a significant reduction in the expression of angiogenesis-related genes (VEGF and VEGFR) in A2058 cells treated with the β-CD-Thymol nanoparticle complex (P < 0.001). The results from the CAM assay further supported these findings. Conclusion: The findings of this study demonstrated a significant decrease in the expression of genes involved in angiogenesis, specifically VEGF and VEGFR, indicating that the β-CD-Thymol complex effectively inhibits angiogenesis in the A2058 skin cancer cell line. Overall, in addition to inhibiting the formation of new blood vessels, the β-CD-Thymol nanostructure shows promise as an anti-angiogenic agent by modulating the expression of VEGF and VEGFR genes.