Platelet-rich fibrin (PRF) has emerged as a promising scaffold for drug delivery, particularly in the context of antimicrobial therapies. This systematic review evaluates the incorporation of antibiotics into PRF to determine its efficacy as a localized antimicrobial delivery system compared to plain PRF without antibiotics. A systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, including 13 in vitro studies with a moderate risk of bias. Antibiotics were incorporated into PRF using different methodologies, including systemic administration before blood collection, addition to blood before centrifugation, and injection into formed PRF matrices. Outcomes were analyzed regarding antibacterial efficacy, structural integrity of PRF, and release kinetics. Antibiotic-enhanced PRF demonstrated significant antibacterial activity against various bacterial strains. The efficacy of the enhanced PRF was dependent on the type of antibiotic, its concentration, and incorporation method. Encapsulation approaches facilitated a sustained antibiotic release, while higher antibiotic concentrations occasionally disrupted PRF integrity. Systemic administration of antibiotics before blood collection enriches PRF effectively, producing significant inhibition zones. The antibacterial effects of PRF outperformed alternative carriers, such as collagen sponges. Antibiotic-loaded PRF is a potent tool for localized antimicrobial delivery, with promising applications in clinical settings. Further research is needed to standardize preparation protocols and explore the impact of different antibiotic delivery methods on PRF's regenerative properties.