Background: Immunization is a key step in preventing the occupational risk of acquiring hepatitis B infection for healthcare workers (HCWs). The aim of the study was determination of hepatitis B vaccine (HepB) coverage rate among HCWs and support staff and immune response 10 years after primary vaccination. Methods: A retrospective study was conducted in the Public Institution Health Centre of Sarajevo Canton in Bosnia and Herzegovina during 2005-2017. Second-generation vaccines Euvax (Sanofi Pasteur, Thailand/LG Life Sciences Ltd, Korea) and Engerix B (GlaxoSmithKline, Belgium) were applied. For quantification of antibody response to hepatitis B virus surface antigen three laboratory methods were used. A level of ≥ 10 IU/L anti-HBs was set as a cut-off value indicating the presence of protective immunity against new HBV infections and successful vaccination. Results: In total, 1541 (75.80%) were fully vaccinated, out of 1126 (73.07%) were females. The median age of participans was 50.5 ± 9.4 years. Control measurement of the anti-HBs level was performed for 409 (26.54%) HCWs, and the presence of protective antibodies was confirmed in 304 (74.33%). During the research, 37 booster doses were administered. Of the 23 retested participants, anti-HBs ≥10 IU/L were observed in 19 (82.16%). Three non-reactors subject were revealed. Conclusion: A satisfying HepB coverage rate and a high protective rate against hepatitis B infection among HCWs and support staff was achived, although vaccination is voluntary in Bosnia and Herzegovina. These results indicate the active engagement of the institution in the implementation of preventive measures and the high level of awareness regarding the significance of immunization. Our results demonstrated the effectiveness of booster doses. A low prevalence of non-reactors was revealed. Additional research with a focus on occupational risk factors in dental service is advised.