Objectives: Aim of this study was to assess and compare health, quality of life, well-being, job satisfaction and job insecurity between nurses, in a tertiary hospital in Greece, working either under permanent or temporary contract. Material and Methods: In this cross-sectional study, consecutively recruited nurses answered a structured questionnaire, the WHO-5 Well-being Index (WHO-5), the Job Insecurity Index (JII), the Work Ability Index (WAI), and the Well-Being at Work Scale (WBWS). Results: Included were 323 nurses (87.6% women, age M±SD 43.68±8.10 years). Tem- porary contract employees had worse quality of life (p = 0.009) and higher job insecurity: both in cognitive dimension (p = 0.013) and emotional dimension (p < 0.001). They also scored worse in the positive affect (p < 0.001), negative affect (p = 0.002) and fulfillment of expectations in work environment (p < 0.001) domains of the WBWS. Additionally, they reported less frequently occupational accidents and injuries (p = 0.001), muscu - loskeletal disorders of the spine or neck (p = 0.007), cardiovascular (p = 0.017), and gastrointestinal (p = 0.010) disorders, while they reported more frequently mental disorders (p < 0.001). Multivariate linear regression analysis showed that temporary work predicted high cognitive (p = 0.010) and emotional (p < 0.001) insecurity, low positive emotions and mood index (p = 0.007), low achievement-fulfillment index (p = 0.047) and high index of negative emotions (p = 0.006), regardless of gender and age. Conclusions: Temporary employment among nurses is associated with a lower sense of job security and well-being, and a higher prevalence of mental disorders, independently of age or gender without a significantly negative effect on their ability to work. Managers, as well as occupational physicians, should recognize the extent of nurses’ job insecurity and assess their ability to work, to provide them with the necessary support and to stimulate the sense of occupational security and work capacity, so that they can thrive in their workplace and therefore be more productive and provide high quality healthcare. Int J Occup Med Environ Health. 2024;37(1):98–109