Aim: We aimed to evaluate if a proactive multidisciplinary diagnostic program could improve outcomes in the management of multidrug resistant (MDR) diabetic foot (DF) osteomyelitis (OM). Methods: We evaluated data of all the consecutive patients who underwent surgical procedure for OM between July and December 2022 in our department (Group A). Intraoperative bone specimens were collected for microbiological and histological analysis on whose results antibacterial therapy was decided. The controls were patients admitted between January and June 2022 with the same indications (Group B), but without systematic intraoperative bone sampling. Clinical and demographic characteristics, procedures, healing rate (HR), healing time (HT) and recurrence rate (RR) were compared between the groups. Results: We derived data from 165 patients: 84 in Group A [50.9%; Male/female 79.3/20.7%; age 70.2±11.2 yrs; Type 1/2 diabetes 6.3/93.7%; diabetes duration (DD) 21.5±12.4 yrs; Hba1c 57.8±19.4 mmol/mol; BMI 27.6±5.1 kg/m2] and 81 in Group B [49.1%; Male/female 82.7/17.3%; age 70.5±11.6 yrs; Type 1/2 diabetes 5.2/94.8%; DD 19.5±11.6 yrs; Hba1c 60.4±21.8 mmol/mol; BMI 27.3±4.8 kg/m2]. Group A had a higher healing rate at six months (84.9% vs 66.9%, p<0.002), a shorter healing time (65±52 days vs 126±92 days, pz<0.001) and a lower recurrence rate at 1 year of follow-up (32.5% vs 58.7%, p<0.05) compared to Group B. In Group A a higher prevalence of infections were polymicrobial (51.1 vs 35%, p<0.002). We observed a higher prevalence of Ps. Aeruginosa (19.7 vs 11.1%, p<0.05) and a lower prevalence of Staph Aureus (22.3 vs 29.6%, p<0.05) in Group A vs Group B. Conclusions: Systematic intraoperative bone sampling for culture and histology, and consequent antibiotic adjustment, increases healing rates and reduces healing times and recurrences rates in DF patients undergoing surgical debridement for OM.