The use of double lumen catheters as a means of hemodialysis access is commonly accompanied with the use of gentamicin as an antibiotic lock. Other antibiotics and anticoagulants are often added to increase the efficacy of gentamicin in order to reduce catheter-related infection and to prevent biofilm formation. This review aimed to evaluate the following: 1) the use of gentamicin in eliminating catheter-related infection and reducing biofilm formation in hemodialysis catheters, 2) the efficacy of additional antibiotics in combination with gentamicin, and 3) the effect of additional anticoagulants to complement the efficacy of gentamicin as the main prophylactic antibiotic lock. We sorted through data from 242 PubMed and ScienceDirect studies, which were then short-listed to 33 studies. Next, they were grouped, extracted, and analyzed qualitatively to fulfil the objectives of this review. Consequently, the use of a gentamicin-lock solution was shown to reduce the incidence of bacteremia; however, it was not strong enough to inhibit the growth of infectious microbes and formation of biofilms. Several bacteria, such as Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Klebsiella pneumoniae, have been reported as infectious agents. Combination with other antibiotics also provided no effect in reducing bacterial growth and biofilm formation in catheters. Furthermore, the additional anticoagulants (trisodium citrate and EDTA) were reported to be effective in enhancing the efficacy of gentamicin in avoiding catheter-related infection, bacterial growth, and biofilm formation; thus, the use of gentamicin can be rationalized.