Cytotoxic drugs used in cancer treatment are inherently toxic due to their genotoxic, teratogenic, and carcinogenic properties. Since healthcare professionals may be exposed to these drugs, it is essential to ensure their safety and to implement exposure assessment programs to document exposure levels, evaluate the effectiveness of protective measures, and establish prevention priorities. This article presents the main results of two exposure assessment campaigns for cytotoxic drugs, conducted in 2010 and 2016 at the same hospital in France. The exposure of healthcare workers (nurses, nursing assistants, and hospital support staff) was assessed using biomonitoring and by measuring contamination in their work environment. The analyses conducted in 2010 revealed the presence of cytotoxic drugs in the urine of nearly half of the nurses and nursing assistants monitored, as well as on work surfaces. Despite the implementation of several preventive measures, the subsequent assessment campaign in 2016 showed that exposures persisted. Analysis of a questionnaire on the overall management of cytotoxic risk shed light on these results. It appears that the various determinants of exposure (training, use of personal protective equipment, incident management, formalized procedures, etc.) must be identified and taken into account to effectively manage cytotoxic risk. Finally, this study highlights the need for regular monitoring of exposures to maintain optimal vigilance among staff and prevent any decline in safety standards over time.