Green nanoparticles are synthesized using environmentally friendly methods, and natural materials hold significant importance. This makes the process environmentally sustainable and reduces the production of harmful waste by-products. Green nanoparticles exhibit reduced toxicity which is crucial for biomedical applications. The current study suggested that yttrium nanoparticles (YNPs) should be synthesized, characterized, and evaluated for their diverse biological applications due to the rise in antibacterial resistance. The YNPs were prepared using a pine needle leaf extract (PNLE). The structural and morphological features have been investigated using X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), photoluminescence (PL), Fourier transform infrared spectroscopy (FTIR), ultraviolet–visible spectroscopy (UV–vis), and vibrating sample magnetometry (VSM). The XRD pattern demonstrated the presence of yttrium oxide and yttrium nitrate phases. The crystallite size and particle size of the synthesized YNPs measured 1.696 nm and 24.55 nm, respectively. The XPS peaks showed two components with binding energies at 530.940 eV and 532.18 eV due to the bond between O–Y and OH–Y, respectively. Additionally, the ferromagnetic nature of the YNPs was confirmed by VSM analysis. The YNPs were tested for antibacterial activity on six uropathogenic bacteria (S. aureus, S. haemolyticus, E. faecalis, E. coli, K. pneumonia, and P. aeruginosa) using the microdilution assays, to find the minimum inhibitory concentration (MIC) as well as the minimum bactericidal concentration (MBC), the agar well diffusion assay, and antibiofilm screening assays, where they showed bacteriostatic action against all isolates (0.5–1 mg/mL MIC) and significant inhibition of biofilm formation (80% inhibition rate). The antioxidant capacity assessed by 1,1, diphenyl-2-picrylhydrazyl (DPPH) radical scavenging revealed 50% DPPH scavenging. Moreover, docking studies exhibited that YNPs inhibit crucial bacterial enzymes, including DNA gyrase, penicillin-binding proteins, carbapenemase, LasR-binding protein, and dihydropteroate synthase. These findings may explain the mechanisms responsible for the observed antibacterial effects of YNPs. Overall, these findings underscore YNPs as promising candidates for antioxidant and antibacterial applications.