Abstract Our aim was to investigate how different oral hard tissues determine the differentiation of osteoclast-like cells. Murine macrophage cells were stimulated for 12 day with RANKL and M-CSF on dentin slices. Morphological changes of cells and hard tissues were examined by electron microscopy and toluidine blue staining. Cells were stimulated with RANKL and M-CSF on pulverized bone, dentin, cementum or polystyrene—with and without stimulation. TRAP staining was performed. To elucidate total gene expression, RNA sequencing was carried out. Four target genes (CXCL2, IGF-1, GDF15, HSPA1b) were selected and their expression was analyzed by RT-PCR and ELISA. Statistics comprised One-way ANOVA and Tukey’s test (P