Objectives <p>To compare the effect of fluocinolone acetonide (FA), mineral trioxide aggregate (MTA), and their combination on inflamed human dental pulp cells (HDPCs), focusing on cell viability, expression of inflammation and mineralisation-associated genes, and odontogenic differentiation.</p> Methods <p>HDPCs were treated with 20&#xa0;µg/mL lipopolysaccharide (LPS) alone or in combination with and 400 µM hydrogen peroxide (H₂O₂) for 6, 12, 24 and 48&#xa0;h to induce inflammatory conditions. Normal and inflamed HDPCs were further treated with FA, MTA extraction medium, or a combination of MTA and FA. Cell viability was assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The expression of inflammatory and mineralisation-associated genes was evaluated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Odontogenic differentiation was determined by alkaline phosphatase staining (ALP) and alizarin red S staining (ARS).</p> Results <p>HDPCs pretreated with LPS and H₂O₂ had reduced cell viability, increased expression of IL-1β and IL-6, decreased expression of ALP and DSPP, and reduced odontogenic differentiation. Treatment with FA alone or in combination with MTA improved these parameters, except for odontogenic differentiation, which was enhanced only in the FA-treated group.</p> Conclusion <p>FA exerted anti-inflammatory properties and promoted odontogenic differentiation under the inflammatory milieus of HDPCs in this <i>in vitro</i> study.</p> Clinical relevance <p>Under the simulated inflammatory condition of HDPCs, this <i>in vitro</i> study suggests the potential benefit of FA for anti-inflammatory and mineralisation enhancing properties.</p>