Therapeutic Potential of Methotrexate-Loaded Superparamagnetic Iron Oxide Nanoparticles Coated with Poly(lactic-co-glycolic acid) and Polyethylene Glycol against Breast Cancer: Development, Characterization, and Comprehensive In Vitro Investigation

Novel superparamagnetic iron oxide nanoparticles (SPIONs)of Methotrexate(MTX) were developed using supercritical liquid technology and optimizedwith a Box-Behnken design in order to assess its potentialas a candidate for the treatment of breast cancer. MTX-SPIONs coatedwith poly(lactic-co-glycolic acid)-polyethyleneglycol 400 had an aggregate size of 500 nm and an encapsulation efficiencyof 46.8 & PLUSMN; 3.9%. The Fourier-transformed infrared spectroscopyanalysis revealed a shift in the main bands due to intermolecularhydrogen bonds, whereas the differential scanning calorimetry analysisrevealed the absence of the MTX melting endotherm, indicating completeencapsulation with oxide nanoparticles. The zeta potential resultsindicated a value of 4.98 mV, whereas the in vitro release study revealedan initial burst release followed by a considerable release of 35.1 & PLUSMN; 2.78% after 12 h. Using flow cytometry, control, MTX, and MTX-SPIONswere evaluated for apoptosis, with MTX-SPIONs exhibiting greater apoptosisthan the control group and MTX. In addition, MTX-SPIONs inhibitedcell division and content organization while substantially increasingthe proportion of cells in the G1 and G2 phases relative to the controlgroup. MTX-SPIONs exhibited prolonged anticancer effects against MCF-7cell lines compared to MTX alone, indicating that SPION-deliveredchemotherapeutics may increase cytotoxicity. The medication was stablewith low encapsulated drug loss, suggesting that the supercriticalliquid technology-based method is a promising way for generating drug-polymermagnetic composite nanoparticles for cancer treatment.