Theaflavin Attenuates Arecoline-induced M1 Polarization and Promotes Homeostasis in Human Periodontal Ligament Fibroblasts

Background

Periodontal Ligament cells (PDL) functions as a bridge between the tooth and bone and playing a vital role in oral health. Periodontitis is an inflammatory condition that leads to dysfunction of Periodontal Ligament (PDL) cells, resulting in pyroptosis. Arecoline is a well-known alkaloid that produce excessive pro-inflammatory cytokines, reactive oxygen species lead to Pyroptosis of PDL cell line. Hence, many bioactive compounds play a vital role on anti-inflammatory activities.

Objectives

This study aimed to the ameliorative effect of Theaflavin on Arecoline induced inflammation of human PDL fibroblasts based on the periodontal regeneration.

Materials and Methods

IC₅₀ of Arecoline, Dexamethasone and Theaflavin, was determined by MTT assay. Inflammation was induced by 64.44 μM of arecoline and further treated with different concentration (1, 5, and 10 μM) of theaflavin and evaluated for its morphological changes. Dexamethasone was used as positive control. To explore the molecular effects, quantitative Real-Time PCR (qRT-PCR) was used to examine the mRNA expression of five key osteogenic and extracellular matrix genes: RUNX2, ALPL, COL1A1, SPP1, BGLAP, with GAPDH serving as the internal control. Statistical analysis was done using one-way ANOVA with Dunnett’s post-hoc test, setting significance at p<0.05.

Results

The Arecoline reduced the viability and osteogenesity of the PDL cells. Theaflavin treatment resulted in the restoration of PDL cells even under Arecoline exposure. Eventually it modified the expression of RUNX2, ALPL, COL1A1 indicating a reactivation of osteogenic differentiation pathways and redox balance.

Conclusion

These findings highlighted TF’s potential as a natural, biocompatible option for supporting periodontal health and countering the oral toxicity associated with arecanut.