In healthy bone, resorption and synthesis are in perfect coordination. In previous studies we demonstrated that the incorporation of silicon into the hydroxyapatite (HA) lattice enhances the proliferation and differentiation of human osteoblasts. Therefore, the aim of this study was to demonstrate the effect of silicon-substituted HA (0.8 and 1.5 wt % Si-HA) on the differentiation of mononuclear cells into osteoclasts, using two different starting cultures, peripheral blood mononuclear cells (PBMC) and monocytes expressing the CD14 antigen (CD14+). Through this study, it was possible to demonstrate that Si-HA allows the differentiation of mononuclear cells into mature osteoclasts, independent of the starting culture, PBMC or CD14+. Most of the cells on the surface of the materials expressed osteoclastic markers: actin rings, several nuclei, positivity for tartrate-resistant acid phosphatase (TRAP), and vitronectin receptor. In the presence of osteoclasts, a higher release of calcium and phosphate into the medium from the 1.5 wt % Si-HA substrate was detected when compared to the HA substrate; therefore, these results indicate higher osteoclastic resorptive activity on the 1.5 wt % Si-HA surface. Si-HA can be resorbed by cellular mechanisms and have a stimulatory effect on osteoclasts, although the underlying mechanism is still poorly understood.