Over the past decades, the world population has become more physically inactive, leading to, obesity, elevated blood pressure and lipid profile, and decreased maximal oxygen uptake (VO2max), which are risk factors for several non-communicable diseases. Health care workers are reported to have an unhealthy lifestyle with a high prevalence of obesity and low VO2max combined with relatively physically demanding work tasks. Female hospital employees were cluster-randomised to either a football group (FG, n=37) or a control group (CG, n=35). FG were encouraged to perform twice-weekly 1-hour training sessions outside working hours for 40 weeks. Based on ITT-analyses after 12 weeks, FG improved (P<0.05) VO2max and fat percentage compared with CG. Furthermore, FG increased (P<0.05) plasma osteocalcin by 21%. After 40 weeks, FG reduced fat mass and increased lower limb bone mineral content (BMC), bone mineral density (BMD) and plasma osteocalcin (P<0.05). Moreover, FG reduced (P<0.05) pain intensity in the neck-shoulder region after 12 and 40 weeks and improved (P<0.05) maximal neck extension strength after 12 and 40 weeks. With regards to long-term compliance, 30% of the original participants have played football together year-round over a 9-year period. The study indicates that football training at a workplace may improve fitness and prevent osteoporosis by promoting physiological health and having preventive effects on future perceived muscle pain in the neck-shoulder region among female hospital employees.