Osteoporosis is a common systemic skeletal disease characterized by reduced bone mass and consequently enhanced susceptibility to fractures. Osteoporotic fractures are associated with increased morbidity and mortality as well as large socioeconomic costs. Several studies have demonstrated that patients receiving long-term glucocorticoid therapy or patients suffering from rheumatoid arthritis (RA) have an increased risk of developing osteoporosis. Therefore, the aim of the present study was to investigate the association between prednisolone therapy, RA and osteoporosis. The study included all patients referred to a dual-energy x-ray absorptiometry (DXA) scan at Aalborg University Hospital during the time period 1st of January 2013 to 31st of December 2013. A total of 3557 patients participated in the study. Data were obtained from a database that contained results from DXA measurements, i.e. T-scores for lumbar spine (L1-L4) and total hip, as well as information about various factors that potentially may affect bone strength. DXA measurements were performed by means of a Hologic Discovery QDR device, whereas the remaining information was obtained from self-completion questionnaires. The results revealed that patients receiving prednisolone therapy had higher T-scores of both lumber spine and total hip compared to non-treated patients. Additionally, no significant correlation between prednisolone dose and T-score was demonstrated. Furthermore, RA patients had higher lumbar spine T-scores compared to patients without RA, whereas total hip T-scores were similar between the two groups. Stepwise multiple linear regression analysis further demonstrated that prednisolone therapy and RA were positive predictors of lumbar spine T-score, whereas they were not predictive of total hip T-score. These results were contrary to the hypothesis as well as the findings of other studies. This was thought to be related to limitations of the present study, including the lack of information regarding treatment time of prednisolone as well as disease duration and severity of RA.