DLX6‐AS1 regulates odonto/osteogenic differentiation in dental pulp cells under the control of BMP9 via the miR‐128‐3p/MAPK14 axis: A laboratory investigation

Abstract Aim The regenerative capacity of dental pulp relies on the odonto/osteogenic differentiation of dental pulp cells (DPCs), but dynamic microenvironmental changes hinder the process. Bone morphogenetic protein 9 (BMP9) promotes differentiation of DPCs towards an odonto/osteogenic lineage, forming dentinal‐like tissue. However, the molecular mechanism underlying its action remains unclear. This study investigates the role of DLX6 antisense RNA 1 (DLX6‐AS1) in odonto/osteogenic differentiation induced by BMP9. Methodology Custom RT 2 profiler PCR array, quantitative Real‐Time PCR (qRT‐PCR) and western blots were used to investigate the expression pattern of DLX6‐AS1 and its potential signal axis. Osteogenic ability was evaluated using alkaline phosphatase and alizarin red S staining. Interactions between lncRNA and miRNA, as well as miRNA and mRNA, were predicted through bioinformatic assays, which were subsequently validated via RNA immunoprecipitation and dual luciferase reporter assays. Student's t ‐test or one‐way ANOVA with post hoc Tukey HSD tests were employed for data analysis, with a p ‐value of less than .05 considered statistically significant. Results DLX6‐AS1 was upregulated upon BMP9 overexpression in DPCs, thereby promoting odonto/osteogenic differentiation. Additionally, miR‐128‐3p participated in BMP9‐induced odonto/osteogenic differentiation by interacting with the downstream signal MAPK14. Modifying the expression of miR‐128‐3p and transfecting pcMAPK14/siMAPK14 had a rescue impact on odonto/osteogenic differentiation downstream of DLX6‐AS1. Lastly, miR‐128‐3p directly interacted with both MAPK14 and DLX6‐AS1. Conclusions DLX6‐AS1 could regulate the odonto/osteogenic differentiation of DPCs under the control of BMP9 through the miR‐128‐3p/MAPK14 axis.