This aimed to study of the use of pumpkin aqueous extract in the preparation of iron nanoparticles, the use of ultraviolet (UV) spectroscopy, atomic force microscopy (AFM), scanning electron microscopy (SEM), infrared spectroscopy (FTIR) and X-ray diffraction (XRD) in characterizing the prepared nanoparticles and determining the active groups, and study of the effect of nanoparticles prepared using pumpkin aqueous extract in treating burns. The nanoparticles were prepared using aqueous extract of pumpkin and iron salts, which are 100% local materials for treating burns and pathogenic microorganisms. As the study showed that iron nanoparticles showed high inhibitory efficacy for the microorganisms under study, which included: Ps. aeruginosa, Pr. Mirabilis, Staph. aureus, E. coli, K. pneumonia. These bacterial types are among the most dangerous bacterial types in the contamination of burns and wounds. As it showed the highest percentage of Ps. Aeruginos, it indicates 42.8%, followed by Staph isolates. Aureus 28.73%, E. coli 12.1%, P. mirabilis, 8.2%, K. pneumoniae8.2%, E. coli 3.37%, respectively. A peak also appeared at the wavelength of 479.2 nm for colloidal solution of nanoparticles prepared using pumpkin aqueous extract and iron salts using a spectrophotometer. The nanoparticles were characterized using SEM, AFM and FTIR. The prepared nanoparticles showed high efficacy in inhibiting the micro-organisms under study when performing MIC and MBC assays. Milliole concentration of nanoparticles also showed high inhibition of all types of microorganisms. When studying the effect of nanoparticles on the burns that may have caused in laboratory mice, a high efficiency in burn restoration and hair growth compared to control groups, treatment with burn ointment and aqueous extract of pumpkin.