BACKGROUND AND OBJECTIVESMyxobacteria have gained recognition as one of the most prolific producers of bioactive secondary metabolites cover a wide range of chemical structures. In regenerative endodontics, odontogenic inducers are an important component of regenerative endodontics and play a significant role in promoting pulp cell differentiation and mineralization. This study aimed to evaluate the effect of myxobacterial metabolites on the differentiation of dental pulp stem cells into odontoblasts.MATERIALS AND METHODSTo obtain metabolites, the myxobacterial strain was inoculated to a ۱/۲ H fermentation medium, and its secondary metabolites were extracted using liquid-liquid extraction. The effect of metabolite exposure on pulp stem cell viability was assessed using the MTT assay. Odontoblastic markers expression was evaluated, and mineralized nodule formation was assessed using alizarin red staining, followed by quantitative measurement of calcium deposition. Additionally, the inhibitory effect of the metabolite extract on oral microbiota the standard broth dilution method (CLSI M۰۷-A۸) was evaluated.RESULTS AND DISCUSSIONThe use of odonto/osteogenic inducers, such as growth factors, is an attractive strategy for promoting ossification and dental tissue repair. However, several challenges exist for the widespread use of growth factors in clinical settings, including their high physiological doses, short half-life, and severe side effects, such as aberrant bone formation, osteolysis, seroma, and retrograde ejaculation. The data showed that myxobacterial extract induces differentiation of dental pulp stem cells by enhancing the expression of Dentin sialophosphoprotein (DSPP) and Dentin matrix protein-۱ (DMP-۱), along with increasing calcium deposition. The results indicated that the nontoxic doses of ۰.۰۹۶ μg/ml and ۰.۰۴۸ μg/ml applied in odontogenic differentiation of stem cells did not inhibit the growth of oral microbiota. This research may lead to the discovery of new sources of medicinal compounds with the potential modality in regenerative dentistry and pulp capping materials for the maintenance of pulp vitality.CONCLUSIONThe data suggest the potential therapeutic effects of myxobacterial secondary metabolites in regenerative endodontics and vital pulp therapy. Considering the importance of the oral microbiome to health, using dentinogenesis inducer compounds that have minimal impact on the normal resident oral microbiome is beneficial. In our follow up studies, the myxobacterial effective compounds having the signaling effect on differentiation will be isolated and characterized for both cytoxicity and inducing effect.