The purpose of the present study was to explore the relation of work-related risk factors and well-being among healthcare workers and the impact on patient safety, using the Health and Work Survey (INSAT) and Mental Health Continuum - Short Form (MHC-SF). A sample of 361 Portuguese healthcare workers participated in this study. The results indicate some significant work-related risk factors: for emotional well-being, Impossible to express myself (β = -0.977), Not having recognition by superiors (β = -1.028) and Have to simulate good mood and/or empathy (β = -1.007); for social well-being, Exposed to the risk of sexual discrimination (β = -2.088), Career progress is almost impossible (β = -1.518), and Have to hide my emotions (β = -2.307); finally for psychological well-being Exposed to the risk of sexual discrimination (β = -2.153), Career progress is almost impossible (β = -1.377), and Have to simulate good mood and/or empathy (β = -3.201). The results showed high levels of well-being despite the exposure of several risk factors at workplace. Regarding the work-related risk factors, the study showed that most of the participants are exposed to several risk factors at workplace (ranging from environmental risk factors, biological to physical), although psychosocial risk factors (work relations with superiors and colleagues, employment relations, and emotional demands) are the ones that most impact on well-being.