Post Doc Medical University, Sofia Sofia, Sofiya, Bulgaria
Abstract:
Objective: Stem cells derived from the apical papilla (SCAP), dental pulp (DPSC), and periodontal ligament (PDL) have demonstrated remarkable potential for proliferation and differentiation when cultured in controlled environments. These human stem cells are recognized as reliable sources for diverse regenerative therapies. However, the impact of prolonged cultivation on their characteristics remains poorly understood. This study aims to bridge this knowledge gap by investigating the effects of extended in vitro cultivation on SCAP, DPSC, and PDL stem cells, with a particular emphasis on identifying markers of cellular senescence.
Methods: Stem cells were isolated from healthy third molars, including SCAP, DPSC, and PDL tissues, and cultured under standard conditions (DMEM with 10% fetal bovine serum) over an extended period. Cultivation spanned nearly four months, encompassing early (1st–3rd), intermediate (10th–12th), and late passages (18th–20th). Cells from these passages were assessed for proliferation, apoptosis, telomerase activity, and beta-galactosidase activity as markers of cellular senescence. Additionally, MTT assays were performed, and HLA expression levels were analyzed.
Results: SCAP, DPSC, and PDL cells were successfully isolated and expanded in vitro. No statistically significant reduction in proliferative capacity was observed between early and late passages (p>0.05). A slight increase in apoptotic cells was detected in late passages, but telomerase and beta-galactosidase activity remained consistent across passages. Similarly, no significant changes in HLA expression were observed among the cell populations.
Conclusion: This study underscores the robust potential of SCAP, DPSC, and PDL stem cells for regenerative medicine. Despite extended cultivation, these cells maintain their proliferative capacity and show minimal signs of senescence, confirming their reliability as candidates for regenerative therapies and tissue engineering applications. These findings provide a foundation for further research and clinical applications involving dental-derived stem cells in regenerative medicine.
Funding Source: Bulgarian National Science Fund МП21/3.4/BG-RRP-2.004-0004-C01
