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Background: There is a positive relationship between mitochondrial damage in the cell and uptake in technetium Tc 99m monomer methoxy isobutyl isonitrile (^99mTc-MIBI) scintigraphy. Severe mitochondrial dysfunction with cell death occurs in patients with diabetic foot ulcers (DFUs). To decide on the level of amputation, ^99mTc-MIBI scintigraphy should be considered. Methods: Prospectively, 24 patients with DFUs were included in the study. Based on treatment that started with the hospitalization, patients were divided into two groups: those whose DFUs healed and did not need surgical intervention (healed group) and those whose DFUs did not regress despite surgical and medical treatment and who required further surgical intervention (reoperation group). Before surgery, ^99mTc-MIBI scintigraphy was performed. The ^99mTc-MIBI uptake rates of the injured foot relative to the healthy foot were recorded. Deep-tissue culture was taken at surgery. Erythrocyte sedimentation rate, white blood cell count, and C-reactive protein (CRP) and albumin levels were measured. Results: The ^99mTc-MIBI uptake rates of patients with poor prognosis were higher at all times than those of patients who did not require revision surgery. A significant difference was found between these values in the 10 and 30 s rates. The mean ± SD CRP level was 86.04 ± 21.87 mg/dL in the healed group and 144.43 ± 27.54 mg/dL in the reoperation group (p = 0.040). There was a positive correlation between ulcerated foot and healthy foot ^99mTc-MIBI involvement rates at 10 and 30 s and CRP values, and a negative correlation between albumin values. Conclusions: There was a significant relationship between ^99mTc-MIBI involvement rates and poor prognosis and reamputation. The correlation between CRP and albumin levels, which are among the predictive values, and ^99mTc-MIBI uptake confirmed this relationship in DFUs, which are difficult to manage and treat.