Introduction: Dental care facilities produce large quantities of Biomedical Waste (BMW) such as surgical needles, wires, extracted teeth, and dental materials that are heavily loaded with blood and saliva. Ineffective waste management increases the health risk to the public, patients, and dental professionals. It also contributes to environmental pollution. Aim of Work: To investigate the knowledge level of BMW, observed proper mercury hygiene, BMW management practice, and associated factors among dentists and dental students at the Faculty of Dentistry, Ain Shams University, Egypt. Materials and Methods: An analytical cross-sectional study was conducted using a standardized self-administered questionnaire distributed among 257 respondents selected by convenience sampling. Results: Only 23 % of participants had a Good knowledge of BMW management and 13.2% had Good BMW management practices. More than half (59.1%) of respondents discarded leftover amalgam scrap incorrectly and the majority (73.5%) did not practice proper mercury hygiene. Regarding the different sociodemographic variables, the educational year was the only significant factor related to the knowledge level (p= 0.03). Higher mercury hygiene practice score was reported among dentists compared to dental students (p=0.02). Higher BMW management practice scores were observed among older participants (p=0.03), with participants with more years of experience (P=0.01) and those with extra work (P=0.02). Lastly, a Good knowledge score was significantly associated with a Good BMW practice score among the studied group. Conclusion and Recommendations: Most of the studied group had poor knowledge and practice of proper mercury hygiene and BMW management. An urgent need for training of dental health personnel on proper BMW handling and disposal is highly warranted.