The bone cement is composed of polymethylmethacrylate powder and liquid methylmethacrylate which is commonly used in clinical applications. Nosocomial pathogens utilize the bone cement surface to induce infections, so that bone cements are loaded with various antibiotics to inhibit the infection. Nevertheless, the bacteria gain resistance against antibiotics since the antibiotics are released at slow rates. Phenolic acids are secondary metabolites of plants to prevent infections; thus, are promising antimicrobial chemicals against pathogens. The main aim of this study is the investigation of the antimicrobial effect of liquid and powder 4-hydroxybenzoic acid-loaded bone cement on Escherichia coli, methicillin-resistant and susceptible Staphylococcus aureus. Previously, liquid and powder vancomycin-loaded bone cement had been studied against MRSA and Candida albicans, caffeic acid-loaded bone cement is also investigated on the S. aureus. This study will be the first investigation that the antimicrobial effect of liquid and powder phenolic acid-loaded bone cement against pathogens. 4-HBA was added directly into polymethylmethacrylate powder and mixed with liquid methylmethacrylate. Cement discs were prepared by metal mold and the antimicrobial assay was carried out to determine the bacterial growth inhibition. The structural features of bone cements such as pores and surface were characterized SEM. The antimicrobial agent release of bone cement was also examined by spectrophotometer and HPLC was used for confirmation. Resistance development against 4-HBA was tested on MRSA and Acinetobacter haemolyticus by exposing them to the sub-inhibitory concentrations. The possible genes for the resistance were checked by ddPCR. The significant inhibition of bacterial growth by 4-HBA-loaded bone cement is expected and liquid 4-HBA is expected to be released at a higher rate. Therefore, the phenolic acids may be used as antimicrobial agents in bone cement to inhibit pathogenic bacterial growth.