The nature of precursor phase during the biomineralization process of bone tissue formation is still controversial. Several phases were hypothesized, among them octacalcium phosphate. In this study, an in situ monitoring of structural changes, taking place upon the octacalcium phosphate bone cement hardening, was carried out in the presence of biopolymer chitosan and simulated body fluid (SBF). Several systems with different combinations of components were studied. The energy dispersive X-ray diffraction was applied to study the structural changes in real time, while morphological properties of the systems were investigated by the scanning electron microscopy. The obtained results evidence that final hydroxyapatite phase is formed only in the presence of chitosan and/or SBF, providing new insights into the in vivo biomineralization mechanism and, consequently, favoring the development of new approaches in biomaterials technology.