<p>The media and temperature chosen for dissolution studies can affect the rate of ion release from bioactive glass–ceramics, play a significant role in the effectiveness in vitro and in vivo, and can alter the antimicrobial properties of the biomaterial. A series of 45SiO₂–14.5NaO₂–14.5CaO–6P₂O₅ wt% sol–gel bioactive glass–ceramics containing 10-wt% zinc (Zn), 6-, 8-, or 9-wt% copper (Cu), and 1-, 2-, or 4-wt% cobalt (Co) were incubated in deionized (DI) water, phosphate buffer saline (PBS), and simulated body fluid (SBF) at room temperature (RT) and in SBF at 37&#xa0;°C. The ion release over time from each glass-ceramics in the various incubation conditions was analyzed using inductively coupled plasma-optical emission spectroscopy (ICP-OES), and the results showed increased release of Zn²⁺, Cu²⁺, and Co²⁺ ions in SBF. The pH of the SBF following incubation with the glass–ceramics had decreased compared to DI water and PBS, but was in the range for cell proliferation for tissue regeneration. Antimicrobial testing against <i>E. coli</i> showed inhibition after 100 and 1000&#xa0;h of ion release from the Cu9–Co1 glass-ceramic, and against <i>C. albicans</i>, all doped glass-ceramics showed inhibition at the same time intervals. The rate of <i>S. aureus</i> growth was slowed in the first 18&#xa0;h of contact in the SBF-incubated doped samples at 37&#xa0;°C. Most notably, the Cu9–Co1 composition showed promising ion release, pH stability, and antimicrobial properties in SBF media at 37&#xa0;°C, which are conditions similar to the environment of the human body.</p> Graphical Abstract <p></p>