IGFBP5 Alleviates Senescence Of Dental Stem Cells For Periodontal Regeneration

This study aims to identify IGFBP5 as a key gene that plays a central role in various models of senescent stem cells and to explore the pathways through which IGFBP5 reverses of senescence and oxidative stress phenotypes, thereby promoting bone regeneration. Senescent cell models were established using hydrogen peroxide treatment and passage to P12. Overexpression of IGFBP5 was achieved through lentiviral-mediated IGFBP5 delivery. Markers were evaluated using SA-β-gal staining to assess senescent cells, and DCFH-DA probes were employed to measure intracellular ROS levels. Osteogenic potential was analyzed via alizarin red and ALP staining. Gene and protein expression were assessed using RT-PCR and western blot analysis. Furthermore, RNA-seq was performed pre- and post- IGFBP5 overexpression to identify changes in gene expression. Two models of dental stem cell senescence were successfully established: oxidative stress-induced senescence (OSS) and replicative senescence (RS). Both exhibited reduced cell viability, increased ROS levels, accumulation of senescent cells, and diminished osteogenic potential. RNA-seq analysis revealed a significant downregulation of IGFBP5 expression. Overexpression of IGFBP5 in senescent stem cells reversed several hallmarks of senescence and oxidative stress. KEGG enrichment analysis indicated IGFBP5 overexpression modulated the expression of genes involved in the Wnt non-canonical signaling pathway. Our findings highlight the pivotal role of IGFBP5 in mitigating cellular senescence in dental stem cells by modulating the Wnt non-canonical pathway. This regulatory effect offers promising therapeutic strategies for bone regeneration, particularly in age-related periodontitis characterized by stem cell senescence and impaired osteogenesis.