This quasi-experimental, pre- and posttest study evaluated the impact of a 9-month collaborative regional evidence-based practice (EBP) fellowship program on practice, attitude, knowledge, and perceived barriers associated with implementation of EBP. Three annual cohorts (N=142) of nurses attending a fellowship program from 2008 to 2010 participated in this study. Paired t tests showed statistically significant increases in practice (+.82; p<.001) and knowledge/skills (+.78; p<.001) associated with EBP, but showed no change in attitude (+.16; p=.198). All four Barriers subscales showed statistically significant decreases (-.10 to -.31; p=.036 to <.001). Hierarchical multiple regression analyses showed that the barriers explained 6.8%, 8.9% and 13.9% of variances in practice, attitude and knowledge/skills, respectively. The collaborative regional fellowship program improved the practice and knowledge/skills associated with EBP. The barriers were significant predictors of the EBP implementation among the targeted group of nurses participating in the fellowship program.