Hydroxyapatite/polyetheretherketone (HA/PEEK) composite materials were prepared via an in situ synthesis process in order to achieve strong bonding between PEEK matrix and hydroxyapatite fillers, and ultimately to improve the mechanical properties of the composites. In the study, the biocompatibility of the synthesized HA/PEEK materials was investigated by acute toxicity test, hemolytic test, sensitization test, pyrogen test, intradermal test, and toxicity assay test on animal tissue and cells for the purpose of examining the possible adverse effects of the residue organic chemicals from the in situ synthesis process. In vivo bioactivity of both lab-synthesized PEEK and HA/PEEK composites with various HA content was also studied. It is found that the in situ synthesized composite materials possess good biocompability without toxicity. Although the bioactivity of the material increases with HA content, the composite material with 5.6 vol % HA exhibits satisfactory bioactivity without compromising its excellent mechanical performance, which hints to a potential use as load-bearing orthopedic material. (C) 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 127: 2581-2587, 2013