Synergistic Effects of Betanin and Thymoquinone Combinations on Periodontal Ligament Cell Proliferation and Migration

Background

Natural polyphenols such as Betanin and Thymoquinone have demonstrated individual therapeutic potential in wound healing, antioxidant activity, and anti-inflammatory responses. However, their combinatorial effects on Periodontal Ligament (PDL) cells remain unexplored. This study investigates the synergistic impact of Betanin and Thymoquinone at different ratios (1:1, 1:2, 2:1) on PDL cell proliferation, migration, and antioxidant capacity.

Materials and Methods

Human PDL cells were cultured and treated with combinations of Betanin and Thymoquinone at five concentrations (5, 10, 50, 75, 100 µg/mL) in ratios 1:1, 1:2, 2:1. MTT assay was used to assess cell viability, while the scratch wound healing assay evaluated cell migration. Antioxidant potential was measured by DPPH radical scavenging assay. Data were collected in triplicates and statistically analyzed using one-way ANOVA followed by Tukey’s post hoc test (p < 0.05).

Results

The 2:1 (Betanin-rich) combination demonstrated the highest cell viability (~98% at 100 µg/mL) and superior wound closure (~61.7%), significantly outperforming other ratios and controls. The 1:1 combination showed moderate effects, whereas the 1:2 (Thymoquinone-rich) group showed comparatively lower responses. DPPH assay confirmed strong antioxidant activity of the 2:1 combination, with ~98% inhibition at the highest concentration.

Conclusion

Betanin and Thymoquinone combinations, particularly at a 2:1 ratio, exhibit synergistic effects in enhancing PDL cell viability, migration, and antioxidant defense. These findings highlight their potential application in periodontal tissue regeneration and natural therapeutic development.