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Bioactive glasses have been used for over three decades in biomedical applications owing to high bioactivity, biocompatibility, as well as the possibility to stimulate regeneration of the bone. The aim of this work was to synthesized bioactive glasses, which contain lithium and strontium, by commercial method melting-quenching, as well as determining the properties, affected by mentioned ions. Differential thermal/thermogravimetric analysis, particle size distribution, energy-dispersive X-ray spectroscopy, inductively coupled plasma optical emission spectrometry, evaluation of the antimicrobial activity, in vitro bioactivity and biocompatibility test and scanning electron microscopy were used for characterization. The results showed that glass transition and crystallization temperatures are decreasing with addition of lithium and strontium ions. Furthermore, the samples showed high inhibition rate of Escherichia coli growth, as well as high bioactivity and biocompatibility. The crystal apatite layer, formed on the surface of synthesized glasses after seven days in simulated body fluid, differs in shape, size and interconnection of the glass particles, which depends on concentration of lithium and strontium ions.