Increasing DNA binding affinity of doxorubicin by loading on Fe₃O₄ nanoparticles: A multi-spectroscopic study

Magnetic Fe₃O₄ nanoparticles were synthesized successfully by co-precipitation method and characterized using XRD, SEM and EDS analyses. Then doxorubicin (DOX, a known anticancer drug) was loaded onto nanoparticles. In vitro DNA interaction of free DOX and loaded DOX onto Fe₃O₄ nanoparticles (DOX-Fe₃O₄) was investigated by DNA-viscosity measurements, UV-visible and fluorescence spectroscopies. The obtained values for binding constant of DOX and DOX-Fe₃O₄ compounds from UV-visible spectroscopies were 0.04 × 10⁵ and 0.68 × 10⁵ L mol^-1, respectively, which confirms DOX-Fe₃O₄ compound have a stronger interaction with CT-DNA compared to DOX. Considerable changes on viscosity of the compounds recommended that their binding mode with CT-DNA is intercalative binding. Fluorescence intensity of DOX and DOX-Fe₃O₄ was quenched via static process by regular addition of CT-DNA. Thermodynamic parameters suggest that Van der Waals forces and hydrogen bonding for DOX and electrostatic forces for DOX-Fe₃O₄ are predominantly responsible for interaction with CT-DNA. Competition fluorescence studies were done by Hoechst 33258 as a well-known groove binder and ethidium bromide (EtBr) as a known intercalator probe. Percentage of displacement for EtBr-DNA complex with DOX and DOX-Fe₃O₄ was 39% and 61%, and for Hoechst-DNA complex was 9% and 5%, respectively. These results confirmed that both compounds are intercalator binders, although DOX-Fe₃O₄ with a further 22% displacement is a stronger intercalator binder than DOX. The stronger interaction of DOX-Fe₃O₄ compared to DOX suggests that the current system can be used as a new and effective way to targeted therapy of anticancer drugs.