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Vitamin D₃ is a hormone involved in the regulation of bone metabolism, mineral homeostasis, and immune response. Almost all dental tissues contain resident mesenchymal stromal cells (MSCs), which are largely similar to bone marrow-derived MSCs. In this narrative review, we summarized the current findings concerning the physiological effects of vitamin D₃ on dental MSCs. The existing literature suggests that dental MSCs possess the ability to convert vitamin D₃ into 25(OH)D₃ and subsequently to the biologically active 1,25(OH)₂D₃. The vitamin D₃ metabolites 25(OH)D₃ and 1,25(OH)₂D₃ stimulate osteogenic differentiation and diminish the inflammatory response of dental MSCs. In addition, 1,25(OH)₂D₃ influences the immunomodulatory properties of MSCs in different dental tissues. Thus, dental MSCs are both producers and targets of 1,25(OH)₂D₃ and might regulate the local vitamin D₃-dependent processes in an autocrine/paracrine manner. The local vitamin D₃ metabolism is assumed to play an essential role in the local physiological processes, but the mechanisms of its regulation in dental MSCs are mostly unknown. The alteration of the local vitamin D₃ metabolism may unravel novel therapeutic modalities for the treatment of periodontitis as well as new strategies for dental tissue regeneration.