Prostaglandin EP1 Receptor Antagonist Improves Human Dental Pulp Stem Cell Functions: A potential Therapeutic for Pulp Regeneration

Background: Human dental pulp stem cells (hDPSCs) are essential for regenerative endodontics due to their multipotency and capacity for neurogenic, odontogenic, and angiogenic differentiation. However, inflammatory mediators such as prostaglandins, which signal through the EP1 receptor, can impair these regenerative functions.

Objective: This study aimed to investigate the effects of a prostaglandin EP1 receptor antagonist on the viability, migration (wound healing), collagen synthesis, and mineralization potential of hDPSCs.

Methods: Commercially available hDPSCs (CLS) were cultured in RPMI medium supplemented with 10% fetal bovine serum and 1% antibiotics under standard culture conditions (37°C, 5% CO₂). Cells were treated with vehicle control or an EP1 receptor antagonist at concentrations of 10 and 200 ng/mL, selected based on preliminary dose-response experiments. Cell viability and migration were evaluated using XTT and wound-healing assays.Osteogenic differentiation was induced using osteogenic medium (confluent hDPSCs cultured for 21 days in osteogenic induction medium containing L-ascorbic acid and sodium glycerophosphate), and collagen synthesis and mineral deposition were assessed by picrosirius red and alizarin red staining, respectively. Matrix composition was further characterized by Fourier-transform infrared (FTIR) spectroscopy, while RNA sequencing was performed to evaluate changes in gene expression.

Results: The EP1 antagonist significantly enhanced cell viability, migration, collagen synthesis, and mineralization in a dose-dependent manner. FTIR analysis confirmed increased protein and mineral matrix deposition. Gene expression analysis indicated notable changes associated with regenerative activity.

Conclusion: Inhibition of the EP1 receptor improves the regenerative potential of hDPSCs, highlighting its promise as a supportive therapeutic approach for pulp regeneration in regenerative endodontics.

Acknowledgements:
This research was supported by NAMA Women Advancement, who served as the strategic partner of the 5th Forum for Women in Research
(QUWA): Together Innovating to Shape the Future at the University of Sharjah.