Multifunctional drug delivery system based on hydroxyapatite may be a research challenge in the treatment and reconstruction of bone tissue. Vitamin D3 has a positive effect on osteogenesis as it increases osteoblast differentiation and mineralization of bone tissue. The purpose of the study presented in this paper has been to examine the possibility of the synthesis of a new multifunctional nanoparticulate system for local delivery of vitamin D3 suitable for applications in bone engineering. The synthesis, characterization and application of three nano particle systems was shown: hydroxyapatite, hydroxyapatite/vitamin D3 and hydroxyapatite/PLGA/vitamin D3. The obtained vitamin D3-loaded nanoparticles based on hydroxyapatite. Characterization was performed with wide-angle X-ray structural analysis (XRD), infrared spectroscopy (FT-IR), field-emission scanning electron microscopy (FESEM), zeta potential (ZP) and particle sized distribution (PSD) methods. Basic in vitro studies were performed implantation of materials in rats. XRD and FT-IR analyses confirmed that the vitamin D3 is loaded hydroxyapatite and PLGA. Different values of zeta potential may refer to different phenomenological processes during in vivo studies