Bio-inspired approaches for mimicking the mineralization process of hydroxyapatite (HAp) with highly regulated structures have been intensively developed using biomacromolecular templates. In the present study, zein, a hydrophobic natural protein, was explored to induce the biomineralization of HAp nanocrystallites on the unique nanofibrous structures generated by electrospinning. Zein electrospun fibers with diameters range from several hundred nanometers down to 100 nm were obtained using formic acid as solvent. Randomly oriented apatite crystals were observed on zein nanofibrous membranes after several incubating cycles. The nature of the deposited mineral phase was confirmed as HAp crystallites by energy-dispersive spectroscopy, Fourier transform infrared spectroscopy, and wide-angle X-ray diffraction. A preliminary study of in vitro cytotoxicity and osteoblast adhesion revealed that mineralized zein nanofibrous membranes did not induce cytotoxic effects and had a positive effect on osteoblast growth. Zein/HAp membranes possessed the unique nanofibrous structural features and coating of HAp nanocrystallites may have a wide variety of potential applications for bone repair and regeneration. POLYM. COMPOS., 34:1163-1171, 2013. (c) 2013 Society of Plastics Engineers