Growing cases of multi-drug resistance (MRS) among microorganisms pathogenic have rendered antibiotics incompetent. For that reason, present research endeavors are focused on devising ground-breaking solutions to this pressing problem, together with the progression of new antibiotics and the increase of their effectiveness. In our study, we produced silver nanoparticles (AgNPs) and antibiotics Amoxicillin (AMO) separately, also, we used a combination of amoxicillin with Ag NPs (AMO@AgNPs) by co-friendly method of Pulsed laser ablation (PLAL). UV/Vi spectrophotometer, XRD, TEM, and FTIR were used to investigate characterizations of Amoxicillin conjugated AgNPs. To evaluated their antibacterial activity, we study activity of silver nanoparticles, Amoxicillin, and their conjugated AMO@AgNPs against two types of human pathogen using a well diffusion method. The results showed AMO @Ag nanoparticles were spherical in shape, with a size distribution between 12 and 97 nm, with an average diameter of 32 nm. Conjugates of AgNPs and amoxicillin have anti-bacterial properties against Streptococcus pyogenes bacteria more than Haemophilus influenzae bacteria. Moreover, the results confirmed that the AMO@AgNPs was more active than either AgNPs alone or Amoxicillin alone. Furthermore, studied on biofilm formation and endotracheal tube effects via AFM thickness measurement evaluation showed properties of AMO@AgNPs in preventing biofilm formation in endotracheal tubes. This study's conclusion emphasizes the creation of Amoxicillin-conjugated silver nanoparticles as a critical strategy for dealing with bacterial strains that are resistant to antibiotics. The study highlights the benefits of AMO-AgNPs as a potentially effective treatment against bacteria. Moreover, our findings suggests that AMO@AgNPs may be useful as breathing tube preservative.