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Periodontitis is a common global disease caused by bacterial dysbiosis leading to tissue destruction, and it is strongly associated with anaerobic bacterial colonization. Therapeutic strategies such as oxygen therapy have been developed to positively influence the dysbiotic microbiota, and the use of oxygen-releasing substances may offer an added benefit of avoiding systemic effects commonly associated with antibiotics taken orally or hyperbaric oxygen therapy. Therefore, the oxygen release of calcium peroxide (CaO₂) was measured using a dissolved oxygen meter, and CaO₂ solutions were prepared by dissolving autoclaved CaO₂ in sterile filtered and deionized water. The effects of CaO₂ on planktonic bacterial growth and metabolic activity, as well as on biofilms of <i>Streptococcus oralis</i> and <i>Porphyromonas gingivalis</i>, were investigated through experiments conducted under anaerobic conditions. The objective of this study was to investigate the potential of CaO₂ as an antimicrobial agent for the treatment of periodontitis. Results showed that CaO₂ selectively inhibited the growth and viability of <i>P. gingivalis</i> (<i>p</i> < 0.001) but had little effect on <i>S. oralis</i> (<i>p</i> < 0.01), indicating that CaO₂ has the potential to selectively affect both planktonic bacteria and mono-species biofilms of <i>P. gingivalis</i>. The results of this study suggest that CaO₂ could be a promising antimicrobial agent with selective activity for the treatment of periodontitis.