Medical University, Sofia MEDICAL UNIVERSITY - SOFIA, Bulgaria
Abstract: This study examines the effects of prolonged in vitro cultivation on SCAP, DPSC, and BMSC stem cells, focusing on their proliferative capacity and cellular senescence markers to evaluate their potential for regenerative medicine. Stem cells from the apical papilla (SCAP), dental pulp (DPSC), and alveolar bone (BMSC) exhibit strong proliferative and differentiation abilities under controlled conditions, making them valuable for regenerative therapies. However, the impact of long-term cultivation on these cells remains unclear. This study aims to assess how extended in vitro culture affects SCAP, DPSC, and BMSC, with an emphasis on cellular senescence markers. Stem cells were isolated from healthy third molars (SCAP, DPSC, and BMSC) and cultured in DMEM with 10% fetal bovine serum for approximately four months. Cultivation was categorized into early (1st–3rd), intermediate (10th–12th), and late passages (18th–20th). Gene expression analysis was performed using the RT2 Profiler PCR Array – PAHS – 178ZE-4 for Human Aging. SCAP, DPSC, and BMSC were successfully isolated and expanded. Gene analysis focused on telomere maintenance, DNA damage response, oxidative stress, mitochondrial function, cellular senescence, apoptosis, inflammation, and metabolic regulation. A slight increase in apoptotic cells was observed at later passages, while telomerase activity remained stable throughout. SCAP, DPSC, and BMSC exhibit strong potential for regenerative medicine. Despite prolonged cultivation, these cells maintained their proliferative capacity and showed minimal signs of senescence, reinforcing their suitability for regenerative therapies and tissue engineering. This study provides a foundation for future research and clinical applications of dental-derived stem cells.
Funding Source: This study is financed by the European Union-NextGeneratio- nEU, through the National Recovery and Resilience Plan of the Republic of Bulgaria, project № BG-RRP-2.004-0004-C01.
