Incubation time affects the clinical features and outcome of many nosocomial infections. However, its role in the setting of post-sternotomy mediastinitis (PSM) has not been specifically studied. The present study aimed to evaluate the impact of time to onset of PSM on the clinical presentation and outcomes of patients. Hospital records of 197 patients who developed PSM over a 10-year period and were treated by closed drainage using Redon catheters were reviewed retrospectively. Follow-up was complete for all included patients (median of 19 months); 98 patients developed early-onset PSM (time from initial operation to PSM <14 days) and 99 patients had late-onset PSM (≥14 days). Patients with late-onset PSM had a higher rate of internal thoracic artery harvest and mediastinal re-exploration after initial operation. Patients with early-onset PSM presented more frequently with septic shock. Microbiological findings differed between early- and late-onset PSM by a higher incidence of Enterococcus species in the former and of Staphylococcus aureus in the latter. Overall mortality reached 34% (n = 66). Rates of superinfection, treatment failure, mediastinitis-related death, mortality at 1 year and overall mortality were all significantly higher in patients with early-onset PSM. Multiple regression procedures identified early-onset PSM as a significant and independent risk factor for both 1-year (OR 2.40; 95% CI 1.12-5.11) and overall (OR 2.11; 95% 1.26-3.53) mortality. In conclusion, the results obtained in the present study support the distinction between early- and late-onset PSM with different clinical and pathophysiological features. Early-onset PSM is associated with a significantly higher morbidity and mortality compared to late-onset PSM.