This study aims to determine the in vitro and in vivo biocompatibility and the differences in LL‐37 releasing patterns of the PLC/Col/LL‐37 sustained releasing membrane and the rapid releasing Col/LL‐37 membrane for use as a barrier membrane in guided bone regeneration. On Days 3 and 7, the PLC/Col/LL‐37 membrane revealed generated osteoclasts, whereas the PLC/Col membrane indicated mature osteoclasts. PLC/Col/LL‐37 was non‐toxic to blood cells and inhibited human albumin adhesion to the surface. A microbial reduction was observed in the Col/LL‐37 membrane, while both PLC/Col/LL‐37 and Col/LL‐37 may have caused early phases of skin sensitization. In animal testing, PLC/Col/LL37 and Col/LL37 exhibited the same levels of inflammation as the control group at all time points, while the different releasing patterns of LL‐37 loaded on PLC/Col/LL‐37 and Col/LL‐37 membranes had no effect on inflammation resolution in rat models.