Neohesperidin exerts subtle yet comprehensive regulation of mouse dental papilla cell-23 in vitro

Objective

The molecular regulation of odontoblasts in dentin formation remains largely uncharacterized. Using neohesperidin (NEO), a well-documented osteoblast regulator, we investigated whether and how NEO participates in odontoblast regulation through longitudinal treatments using various doses of NEO.

Design

Mouse dental papilla cell-23 (MDPC-23) served as a model for odontoblasts. MDPC-23 were treated with various doses of NEO (0, 1, 5, 10, 15, 20 μmol/L). Proliferation was assessed using the Cell counting kit-8 assay. Survival/apoptosis was assayed by live/dead ratio. Migration capability was assessed using scratch healing and Transwell migration assays. Mineralization was assessed using alkaline phosphatase staining and alizarin red staining. The expression levels of four key genes (Runx2, osteocalcin [OCN], β-catenin, and bone morphogenetic protein [BMP]−2) representing NEO-induced differentiation of MDPC-23 were measured by quantitative reverse transcription polymerase chain reaction.

Results

The proliferation trajectories of MDPC-23 treated with the five doses of NEO demonstrated similar curves, with a rapid increase in the 10 μmol/L NEO condition after 48 h of treatment. Similar dose-dependent trajectories were observed for survival/apoptosis. All four key genes representing odontogenic differentiation were upregulated in MDPC-23 induced by NEO treatments at two optimal doses (5 μmol/L and 10 μmol/L). Optimal migration and mobility trajectories were observed in MDPC-23 treated with 10 μmol/L NEO. Optimal mineralization was observed in MDPC-23 treated with 5 μmol/L NEO.

Conclusion

NEO can subtly regulate odontoblast proliferation, differentiation, migration, and mineralization in vitro. NEO at 5–10 μmol/L offers a safe and effective perspective for clinical promotion of dentin bridge formation in teenagers.