Yeasts, especially Saccharomyces cerevisiae, are organisms that are essential for fermentation processes but are also used for scientific purposes. One of the strains commonly used in scientific research is Saccharomyces cerevisie W303. Yeast mitochondria play a key role in energy production, and monitoring changes in the mitochondrial proteome (from all proteins extracted) provides good insight into the functionality of these organelles. Two-dimensional (2D) electrophoresis is commonly used to study mitochondrial proteomes. The aim of this work was to analyze the mitochondrial 2D proteome of the yeast S. cerevisie W303 treated with different amounts and types of metal ions eluted from dental appliances. Dental or orthodontic appliances are the most commonly used form of orthodontic therapy today and consist of 2 archwires, 20 brackets, 20 ligatures, and 4 bands. Eluates obtained by eluting metal ions from such dental appliances over a period of 3, 7, and 14 days into a yeast growth medium were used to grow yeast to early exponential phase and produce fractions of mitochondria and proteins, respectively. 2D electroferograms of the treated mitochondria were processed and compared using Dymension software (SyngeneTM Dymension 2), while the content and composition of metal ions in the precipitates were determined using the ICP-MS method. The ions Fe, Ni and Cr predominated in the extracts. Their content increased with the duration of elution and was highest after an elution period of 14 days. 2D electrophoresis showed that treatment of yeasts with eluates leads to changes in mitochondrial proteins, especially those involved in electron transport, ATP synthesis and combating oxidative stress.