Abstract
BACKGROUND: Wound healing is a highly coordinated biological process involving inflammation resolution and neovascularization. Dysregulation of these processes can result in chronic wounds, especially in inflammatory or oxidative stress-rich environments. There is growing interest in natural compounds that can simultaneously promote angiogenesis and suppress inflammation to enhance wound healing outcomes. OBJECTIVE: This study aims to evaluate the synergistic wound-healing effects of betanin and theaflavin in vitro by assessing their ability to enhance cell viability, migration, and gene expression related to inflammation and angiogenesis in human periodontal ligament (PDL) fibroblasts. METHODS: PDL fibroblasts were treated with betanin, theaflavin, and their combinations at two concentrations (viz., 1, 5, 10, 25, 50, 75, 100, 125 μg/mL) were preliminary screened by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and based on the IC(50) values, further tested with two concentrations (10 μg/mL and 25 μg/mL, individually and in combination). The scratch assay was employed to evaluate fibroblast migration. To assess molecular changes, qRT-PCR was performed to quantify expression levels of VEGF-A, HIF-1α, NF-κB, IκBα, and IL-10. RESULTS: Combination treatment significantly enhanced cell viability and wound closure compared to individual treatments and controls. Gene expression analysis showed increased expression of VEGF-A and HIF-1α, indicating improved angiogenic potential. Concurrently, NF-κB expression was reduced, while IκBα and IL-10 were upregulated, indicating anti-inflammatory activity. CONCLUSION: The combined use of betanin and theaflavin enhances in vitro wound healing by promoting angiogenesis and modulating inflammatory responses. This integrative strategy may offer a promising natural therapeutic approach for managing tissue repair in oral and dermal wounds.