Evaluation of Self-Regulating Doped Ferrite Nanoparticles with Glucose, Chitosan, and Poly-Ethylene Glycol Coatings for Hyperthermia and Dual Imaging

Alessandro Negri,1 Anita Conti,1 Emil Milan,2 Enrico Forlin,3 Filippo Gherlinzoni,4 Giovanni Morana,4 Michele Gottardi,4 Paolo Matteazzi,3,4 Adolfo Speghini,2,5 Andre Bongers,6 Pasquina Marzola1,5 1Department of Engineering for Innovation Medicine, University of Verona, Verona, Italy; 2Nanomaterials Research Group, Department of Biotechnology, University of Verona, Verona, Italy; 3M.B.N. Nanomaterialia S.p.A., Carbonera, Treviso, Italy; 4Foundation for Nanotheranostics Research in Cancer Therapy, Treviso, Italy; 5National Interuniversity Consortium of Materials Science and Technology, Firenze, Italy; 6Biological Resources Imaging Laboratory, Mark Wainwright Analytical Centre, UNSW Sydney, Sydney, NSW, AustraliaCorrespondence: Pasquina Marzola, Department of Engineering for Innovation Medicine, University of Verona, Strada le Grazie – 15, Verona, 37134, Italy, Email [email protected]: This study investigates the theranostic potential of doped ferrite nanoparticles (NPs) with self-regulating temperature (SRT) properties, termed M55, coated with glucose (GM55), chitosan (CM55), and poly-ethylene glycol (PM55). The NPs were assessed for their physicochemical attributes, magnetic fluid hyperthermia (MFH) efficacy, dual-imaging capabilities in Magnetic Resonance Imaging (MRI) and Magnetic Particle Imaging (MPI), cytocompatibility, and cellular uptake.Methods: Physicochemical characterization was conducted using Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), and zeta potential measurements. The biocompatibility and cellular uptake were evaluated in MDA-MB-231 breast cancer cells, and MFH performance was tested in vitro. Following intravenous administration, MRI and MPI functionalities were analyzed through phantom studies and in vivo murine models.Results: Coated M55 NPs displayed high colloidal stability in water and effective functionalization. The specific absorption rate (SAR) of 24.4 ± 1.4 W/g confirmed their suitability for MFH applications. In vitro assays indicated excellent biocompatibility and substantial cellular internalization, with GM55 showing the highest uptake and MFH efficiency, reducing cell viability to 50.62 ± 3.92% post two treatment cycles, compared to 67.71 ± 6.11% (CM55) and 71.39 ± 5.84% (PM55). MRI transverse relaxivity (r2) values were notably high across all coatings, enhancing imaging contrast. MPI analysis demonstrated superior cell labeling sensitivity, with GM55 achieving the most pronounced detection. In vivo imaging confirmed effective NPs accumulation in the liver, underscoring their utility as dual MRI/MPI contrast agents (CAs).Conclusion: Coated M55 NPs exhibit significant promise as multifunctional theranostic agents for cancer treatment. GM55, in particular, offers superior MFH efficacy and cellular uptake, while CM55 and PM55 may present unique advantages for alternative biomedical applications. The dual-imaging capabilities of these NPs provide a robust platform for real-time monitoring of distribution and therapeutic outcomes. Future investigations will focus on optimizing NPs formulations and expanding in vivo assessments to advance clinical translation.Keywords: nanotheranostics, magnetic fluid hyperthermia, magnetic resonance imaging, magnetic particle imaging