Theaflavin-Loaded Zinc Nanoparticles Enhance Epithelial Cell Migration Through Activation of the Notch1 Signalling Pathway

Abstract

Background: Periodontal regeneration necessitates biomaterials which possess optimal biocompatibility, anti-inflammatory, and pro migratory properties. Theaflavin, a potent polyphenol which is used as medicine in heart health, cardiovascular diseases etc and zinc oxide nanoparticles are recognized individually for their bioactive functions. This research investigates the combined effect of Theaflavin derived zinc oxide nanoparticles (TheoZnONPs) on human periodontal ligament cells.

Methods: TheoZnONPs were produced and evaluated using conventional techniques. Biocompatibility was assessed via MTT assay at various concentrations (5-200 µg/mL). Anti-inflammatory activity was assessed by inhibiting protein denaturation and antioxidant potential was evaluated using DPPH radical scavenging assay. Cell migration was evaluated through an in vitro scratch assay over a 24 hour period. Quantitative real time PCR assessed the expression level of Notch pathway genes (Notch1, Jagged 1, Hes 1, DLL4) and VEGF-A, using Gapdh as a housekeeping gene.

Results: TheoZnONPs showed enhanced biocompatibility, maintaining over 80% viability at therapeutic doses. Anti-inflammatory and anti-oxidant activities were notably greater as compared to theaflavin or zinc oxide nanoparticles alone with >94% inhibition of protein denaturation and ~97% DPPH scavenging at 200 µg/mL. The scratch assay revealed enhanced cell migration with 78.6% wound closure at 24 hours. Gene expression analysis showed substantial upregulation of Notch 1, Jagged 1 and VEGF-A, suggesting the activation of regenerative pathways.

Conclusion: TheoZnONPs combine the bioactivities of theaflavin and ZnONPs, enhancing PDL cell viability, migration and gene expression linked with wound healing. These results support their potential application in periodontal tissue regeneration and advanced wound care.