Dual Regulation of Osteogenesis and Inflammation by Pomegranate (Punica granatum L.) Extract in Periodontal Ligament-Derived Stem Cells: Implications for Regenerative Medicine

Document Type

Article

Publication Date

Winter 1-23-2026

Abstract

Objectives

This article aims to compare the osteogenic along with anti-inflammatory activity of different concentrations of pomegranate extract (PG) on human periodontal ligament-derived stem cells (hPDLSCs) in vitro.

Materials and Methods

hPDLSCs were acquired from the root surface of removed molars, cultured, and then distinguished by flow cytometry analysis and several lineage differentiation potentials. To identify the effect of PG on hPDLSCs' viability, an MTT assay was performed. hPDLSCs were maintained in osteogenic induction medium with varying concentrations of PG. At the end of the induction period, osteogenesis was assessed using Alizarin Red staining, ALP assay, and qRT-PCR to determine the expression of OPG, RUNX2, Ki67, and TNF-α.

Statistical Analysis

All experiments were conducted in triplicate, and data are presented as the mean ± standard deviation (SD). One-way ANOVA followed by Tukey's post hoc test was performed to assess statistical significance at a threshold of p <  0.05.

Results

The results of the MTT assay demonstrated that 100 mg/mL PG had significantly lower cell viability than the other concentrations. Statistical analysis of the ALP enzyme activity was mostly pronounced at 6.25 and 12.5 mg/mL concentrations, while it was least pronounced at 50 mg/mL. PCR revealed that the group treated with a 6.25-mg/mL concentration exhibited significantly elevated expression rates of RUNX2, OPG, and Ki67 in contrast with the control group. Conversely, the 50-mg/mL concentration group demonstrated the lowest expression levels. Regarding TNF-α, the 50-mg/mL concentration group showed the greatest expression levels compared with the control group and all other concentrations.

Conclusion

The data indicate that low concentrations of PG could enhance osteogenic differentiation and exert anti-inflammatory effects on hPDLSCs. These dual actions suggest that PG, at optimized doses, may serve as a promising natural agent for periodontal regeneration and bone tissue engineering. Future studies are warranted to evaluate its clinical potential in regenerative medicine.

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