Abstract
OBJECTIVE: This study investigates the regenerative and protective effects of postbiotics (cell-free supernatant) derived from the Lactiplantibacillus plantarum EIR/IF-1 strain on human periodontal ligament mesenchymal stromal cells (hPDL-MSCs). MATERIALS AND METHODS: hPDL-MSCs were isolated from periodontal ligament tissues (PDL) of wisdom teeth using enzymatic digestion and subsequently characterized through immunophenotyping. The effect of postbiotics on the viability of hPDL-MSCs was assessed using the MTT assay and flow cytometry, while their impact on cell migration was evaluated via the scratch assay. Anti-inflammatory effects of postbiotics were investigated on lipopolysaccharide (LPS, derived from Porphyromonas gingivalis)-stimulated hPDL-MSCs through Enzyme-Linked Immunosorbent Assay (ELISA). Additionally, the antioxidant effects of postbiotics were analyzed in hydrogen peroxide (H₂O₂)-induced hPDL-MSCs by measuring reactive oxygen species (ROS) levels using flow cytometry. The expression of collagen type I (COL1A1) gene was further assessed by quantitative reverse transcription PCR and immunofluorescence staining. RESULTS: Treatment with postbiotics (250 µg/mL) significantly increased the viability and migration capability of hPDL-MSCs, while enhancing collagen production for PDL repair. Treatment with postbiotics for 24 h resulted in a 54.53 ± 2.01% reduction in intracellular ROS levels compared to untreated H(2)O(2)-induced hPDL-MSCs. Furthermore, postbiotics significantly decreased the production of pro-inflammatory cytokines (IL-8, IL-6, and IL-1β), and increased the anti-inflammatory cytokine IL-10 (2.67-fold) compared to untreated LPS-stimulated hPDL-MSCs. CONCLUSION: Our findings indicate that postbiotics exhibit biological activity throughout all stages of the healing process, beginning with the modulation of the inflammatory response to LPS stimulation, followed by the promotion of cell migration, proliferation, and collagen synthesis. Given the unmet need for safe and adjuvant therapeutic approaches that promote comprehensive periodontal regeneration in periodontal diseases, this study presents postbiotics as a promising candidate. CLINICAL RELEVANCE: Postbiotics could be integrated into regenerative therapies as a novel bioactive material to improve the healing and regenerative outcomes in periodontal defects by both controlling inflammation and stimulating tissue repair processes.