The increasing number of antibiotic-resistant bacteria has become a challenge for contemporary medicine. Nanotechnology brings new possibilities in effectively combating multi-resistant bacteria, in which one of the most promising system is the silver nanostrutuctures (SNS) that can be associated with polymers for biomedical use. Herein, Pluronic F127 was functionalized with amino acids in obtaining and stabilizing of SNS. The amino acids glycine, L-Alanine and DL-Alanine were used in the esterification reaction of the PEO end groups present in the polymer chain. These polymers were used in the reduction of Ag⁺ ions and in the stabilization of the nanoparticles by the hydrothermal and the photosensitization routes. The functionalized polymers were characterized by FTIR, in which esterification probably occurred since bands of amine groups was observed, while the stretching band of carbonyl groups from amino acids (-C=O) was absent. SNS were characterized by UV-Vis absorption spectroscopy and by TEM analyze. The SNS obtained by the hydrothermal route are predominantly spherical, whereas those obtained by photosensitization give rise to rods and prisms. Analyses by Fluorescence confirmed that metal clusters were present in the samples obtained by photo-assisted method using glycine and D.L-alanine. The SNS obtained by photo-assisted route were efficient against S. aureus, evaluated by the well diffusion method, in which the ones containing clusters presented larger zones of inhibition.