Bioactive glasses undergo dynamic changes in vivo to produce an apatite layer permitting a strong bond with living tissues including both bone and soft tissues, and their compositions can be modified and tailored. The aim of this project was to generate high phosphate low fluoride containing bioactive glasses and explore their bioactivity and biological performances in vitro. Bioactive glasses (0-7% F- content, constant 6.33% P2O5 in Mol.%) were produced and the particles immersed in Tris Buffer solution or cell culture medium (α-MEM) to determine apatite formation and ion (Ca, P, Si and F) release. Bioactive glass conditioned medium was used to treat pre-osteoblasts MC3T3-E1 for cytotoxicity, pre-osteogenic and pro-angiogenic responses, and to human oral fibroblasts and epithelial cells for proliferation. Antibacterial ability was explored by incubating supra- and sub-gingival bacteria with bioactive glass particulates. Rapid apatite formation was observed in F- containing bioactive glasses after only 2 h immersion in Tris buffer solution, while it was not detectable until 72 h in the F- free bioactive glass. Alkaline phosphatase activity, cell number, collagen formation, bone-like mineral nodules and osteogenic gene expression of MC3T3-E1 cells were significantly promoted in low F- bioactive glass (P6.33F1) conditioned medium. MC3T3-E1 VEGF gene expression was increased, and protein production was dose-dependently promoted with F- containing bioactive glass conditioned medium, which also promoted human oral fibroblast proliferation, but suppressed epithelial cell numbers. After incubation with glass particulates, the growth of L. casei, S. mitis, A. actinomycetemcomitans and P. gingivalis, was significantly inhibited; the antibacterial activity being dependent on the F- content of the bioactive glasses. As a potential bone graft substitute in vivo, such novel bioactive glasses would be expected to stimulate bone formation and overcome problems associated with infection and the poor vascularisation in large bone graft sites. Additionally, they could reduce the need for further clinical intervention, and in particular, will be advantageous for the periodontal soft tissue regeneration.