The easy clinical handling and applicability of biomaterials has become a focus of materials research due to rapidly increasing time and cost pressures in the public health sector. The present study assesses the in vitro and in vivo performance of a flexible, mouldable, cottonwool-like nanocomposite based on poly(lactide-co-glycolide) and amorphous tricalcium phosphate nanoparticles (PLGA/TCP 60:40). Immersion in simulated body fluid showed exceptional in vitro bioactivity for TCP-containing fibres (mass gain: 18%, 2 days, HAp deposition). Bone regeneration was quantitatively investigated by creating four circular non-critical-size calvarial defects in New Zealand White rabbits. The defects were filled with the easy applicable cottonwool-like PLGA/TCP fibres or PLGA alone. Porous bovine-derived mineral (Bio-Oss) was used as a positive control and cavities left empty served as a negative control. The area fraction of newly formed bone (4 weeks implantation) was significantly increased for TCP-containing fibres compared to pure PLGA (histological and micro-computed tomographic analysis). A spongiosa-like structure of the newly formed bone tissue was observed for PLGA/TCP nanocomposites, whereas Bio-Oss-treated defects afforded a solid cortical bone.