This study was set up to evaluate the costs of a one-stage treatment of chronic osteomyelitis using bioactive glass S53P4 versus a two-stage treatment using gentamicin-loaded PMMA beads. Furthermore, a cost-effectiveness analysis was performed from a hospital's perspective together with the evaluation of clinical outcome. A treatment group (n = 25) receiving one-stage surgery with bioactive glass was retrospectively compared with a two-stage control group (n = 25). An assessment was made of all costs included from first outpatient visit until one year after treatment. Bootstrap simulation and sensitivity analyses were performed. The primary endpoint was cost-effectiveness with clinical outcome as the secondary endpoint. The base case analyses shows dominance of the one-stage treatment with bioactive glass S53P4 due to lower costs and a better clinical outcome. Sensitivity analyses confirm these findings. This study is the first in its kind to show one-stage treatment of chronic osteomyelitis with bioactive glass S53P4 to be cost-effective.