Iron oxide nanoparticles (Fe₂O₃ NPs) were successfully Synthesized in a green manner using Cucurbita pepo pulp extract as a natural capping and reducing agent. UV-vis spectroscopy verified the synthesis with a distinct absorption peak at 285 nm, while FTIR analysis revealed functional groups responsible for reduction and stabilization. X-ray diffraction (XRD) analysis confirmed the crystalline nature of the nanoparticles, with an average size of 21.5 nm. SEM and EDX analyses further validated the nanoparticles' spherical morphology and elemental composition. Biosynthesized IONPs exhibited notable antibacterial activity against multidrug-resistant bacterial strains such as Klebsiella pneumoniae and Pseudomonas aeruginosa. The inhibition zones ranged between 5-22 mm for Klebsiella pneumoniae and from 4 to 12 mm for Pseudomonas aeruginosa, depending on the concentration of the nanoparticles. Hematological evaluations demonstrated strong anticoagulant and thrombolytic properties. Iron oxide nanoparticles effectively inhibited blood coagulation at 40 μg/mL and showed significant thrombolytic activity by dissolving preformed clots at 50 μg/mL. The biosynthesized IONPs showed remarkable antioxidant activity that was comparable to standard. This study underscores the potential of Cucurbita pepo as a sustainable and eco-friendly resource for synthesizing multifunctional IONPs. The results suggest promising applications to address antibiotic resistance and manage blood-related disorders. Furthermore, the findings highlight the critical role of green nanotechnology in the advancement of environmentally sustainable and biocompatible nanomaterials for diverse biomedical applications.