Received January 21, 2026
Accepted February 18, 2026

Objective: To evaluate the migration and cell viability of fresh and frozen seminal plasma at three different concentrations on periodontal ligament fibroblasts. Methods: Semen samples from three healthy volunteers were processed to obtain seminal plasma, filtered (0.22 µm), and diluted to 2.5% and 5% (v/v) in DMEM. Aliquots were tested fresh or after storage at -20°C for 8 days. Cell migration was evaluated by assessing wound closure (wound-healing) using photographs analyzed with ImageJ. Cell proliferation was evaluated using the commercial Cell Titer 96® Aqueous Non-Radioactive Cell Proliferation Assay kit in triplicate at 6, 12, and 24 hours. Results: Across donors, frozen seminal plasma showed a higher mean percentage wound closure at 48 h than fresh seminal plasma; however, differences versus controls were not consistently significant, and viability tended to decrease with seminal plasma exposure, particularly after freezing Conclusion: Fresh and frozen seminal plasma do not promote cell migration and proliferation at the concentrations used. The apparent trend toward greater migration with frozen samples requires confirmation in adequately powered studies and should be interpreted cautiously.