This study evaluated the osteogenic differentiation of human dental pulp stem cells in response to substances released by the pulp capping agents, Biodentine (BD), mineral trioxide aggregate (MTA) and two-paste calcium hydroxide cement (CHC), along with their physicochemical characteristics. The dimensional stability test showed that of the materials studied, only BD met the standards recommended by the International Organization for Standardization (ISO) for pulp capping materials and thus can be used safely. In the chemical tests, BD was the most stable material. In the Alizarin red S test, BD formed the higher amount of mineralized nodules in the mineralizing medium and also formed mineralized nodules in a non-mineralizing medium. BD releases substances that can significantly induce formation of the human dental pulp stem cell-mineralized extracellular matrix, with physicochemical characteristics that are more conducive to pulp repair than those of MTA and CHC.