Multimodal Evaluation of Punica granatum and Terminalia chebula Seed Extract–Loaded Biogel on Implant Surface Preservation, Biofilm Virulence, and Osteogenic Differentiation: An In Vitro Study

Abstract

Objectives: This study evaluated the antibiofilm activity, titanium surface compatibility, and osteogenic potential of a thermosensitive biogel containing Punica granatum seed extract (PGSE) and Terminalia chebula seed extract (TCSE), compared with chlorhexidine (CHX).
Materials and Methods: Grade IV titanium discs with Staphylococcus aureus biofilms were treated with control, 0.2% CHX, TCSE, PGSE, or PGSE+TCSE biogels. Antibiofilm activity was assessed by extracellular polymeric substance (EPS) quantification and confocal laser scanning microscopy. Titanium surface characteristics were evaluated by scanning electron microscopy, surface roughness, and contact angle analysis. Osteogenic activity was investigated in MC3T3-E1 cells using alkaline phosphatase (ALP) activity and Alizarin Red S staining. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test (p < 0.05).
Results: PGSE+TCSE showed the greatest EPS reduction (72.2%), lowest biofilm thickness (4.6 ± 0.7 µm), and highest bacterial death (91.3%). It maintained favorable titanium surface characteristics, with roughness of 0.28 µm and contact angle of 31.7°. The formulation also produced the highest ALP activity (3.28 ± 0.20-fold) and mineralization (3.12 ± 0.18-fold), significantly higher than CHX and individual extracts (p < 0.001).
Conclusion: PGSE+TCSE thermosensitive biogel demonstrated promising antibiofilm, titanium surface-preserving, and osteogenic properties, supporting its potential as a local adjunct for peri-implantitis management.