AIM:The purpose of the paper was to assess job demands, burnout, and teamwork in healthcare professionals (HPs) working in a general hospital that was analysed at two points in time with a time lag of three years.METHODS:Time 1 respondents (N = 325) were HPs who participated during the first wave of data collection (2011). Time 2 respondents (N = 197) were HPs from the same hospital who responded at Time 2 (2014). Job demands, burnout, and teamwork were measured with Hospital Experience Scale, Maslach Burnout Inventory, and Hospital Survey on Patient Safety Culture, respectively.RESULTS:Significantly higher scores of emotional exhaustion (21.03 vs. 15.37, t = 5.1, p < 0.001), depersonalization (4.48 vs. 2.75, t = 3.8, p < 0.001), as well as organizational (2.51 vs. 2.34, t = 2.38, p = 0.017), emotional (2.46 vs. 2.25, t = 3.68, p < 0.001), and cognitive (2.82 vs. 2.64, t = 2.68, p = 0.008) job demands were found at Time 2. Teamwork levels were similar at both points in time (Time 1 = 3.84 vs. Time 2 = 3.84, t = 0.043, p = 0.97).CONCLUSION:Actual longitudinal study revealed significantly higher mean values of emotional exhaustion and depersonalization in 2014 that could be explained by significantly increased job demands between analysed points in time.