EXPLORING BETANIN’S ANTIANGIOGENIC AND PROHEALING EFFECTS IN DENTAL TISSUE REGENERATION EVIDENCE FROM ZEBRAFISH AND CAM MODELS

Background: Regeneration of dental and periodontal tissues requires a coordinated interplay between angiogenesis,
inflammation control, and extracellular matrix remodelling. Tissue repair can be impeded by excessive or unregulated
vascularization and inflammation. Betanin, a natural pigment derived from beetroot, has shown promising antioxidant, anti-inflammatory, and wound-healing effects. However, its role in modulating angiogenesis and promoting regeneration in dental tissues is yet to be explored.
Objectives: To investigate the antiangiogenic and prohealing potential of betanin in dental tissue regeneration using
zebrafish fin regeneration and chick chorioallantoic membrane (CAM) models, and to elucidate its effects on the
expression of key genes associated with angiogenesis, inflammation, and tissue remodelling.
Methods: Zebrafish were subjected to caudal fin amputation and treated with betanin at concentrations of 5 and 10
µg/mL for 7 days. Fin regrowth was assessed macroscopically and histologically. In parallel, CAM assays were
performed by applying betanin (1, 5, 10, 25, 50 µg/egg) on day 7 of incubation. After 48 hours, branching, vascular
density, avascular zone formation, and vessel count were quantified. Gene expression analysis of BMP2, FGF2,
VEGF-A, MMP9, HIF-1α, IL-6, and TNF-α was conducted using qRT-PCR on harvested fin and CAM tissues.
Results: Fin regeneration was enhanced by Betanin therapy in zebrafish, with improved tissue architecture and
vascular organization. In CAM assays, higher concentrations of betanin (25 and 50 µg) significantly reduced blood
vessel branching and density while increasing avascular zone size, indicating a concentration-dependent
antiangiogenic effect. Gene expression analysis revealed upregulation of regenerative markers BMP2 and FGF2, and
downregulation of VEGF-A, HIF-1α, MMP9, IL-6, and TNF-α, which suggested betanin promotes healing while
inhibiting excessive inflammation and angiogenesis.
Conclusion: Betanin exhibits dual properties entailing pro-regenerative and antiangiogenic effects by enhancing tissue repair and regulating angiogenic and inflammatory pathways. These findings support its potential application in dental tissue engineering and wound healing formulations.