title: Drug-induced premature senescence model in human dental follicle stem cells.
abstract: Aging is identified by a progressive decline of physiological integrity leading to age-related degenerative diseases, but its causes is unclear. Human dental pulp stem cells (hDPSCs) has a remarkable rejuvenated capacity that relies on its resident stem cells. However, because of the lack of proper senescence models, exploration of the underlying molecular mechanisms has been hindered. Here, we established a cellular model utilizing a hydroxyurea (HU) treatment protocol and effectively induced Human dental pulp stem cells to undergo cellular senescence. Age-related phenotypic changes were identified by augmented senescence-associated-β-galactosidase (SA-β-gal) staining, declined proliferation and differentiation capacity, elevated G0/G1 cell cycle arrest, increased apoptosis and reactive oxygen species levels. Furthermore, we tested the expression of key genes in various DNA repair pathways including nonhomologous end-joining (NHEJ) and homologous recombination (HR) pathways. In addition, our results showed that Dental pulp stem cells from young donors are more resistant to apoptosis and exhibit increased non-homologous end joining activity compared to old donors. Further transcriptome analysis demonstrate that multiple pathways are involved in the HU-induced Dental pulp stem cells ageing, including genes associated with DNA damage and repair, mitochondrial dysfunction and increased reactive oxygen species levels. Taken together, the cellular model have important implications for understanding the molecular exploration of Dental pulp stem cells senescence and aging.
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.
authors.forename: Yuanfen
authors.initials: Y
authors.lastname: Zhai
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.
authors.forename: Rongbin
authors.initials: R
authors.lastname: Wei
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.
authors.forename: Junjun
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Pediatric Dentistry, School of Stomatology, Tongji University, Shanghai Engineering Research Center, Shanghai, P. R. China.
authors.forename: Huihui
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.
authors.forename: Wenping
authors.initials: W
authors.lastname: Cai
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.
authors.forename: Yongguang
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Stomatology, Huashan Hospital, Fudan University, Shanghai, P. R. China.
authors.forename: Shuwei
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.
authors.forename: Tianshu
authors.initials: T
authors.lastname: Yang
authors.affiliation: Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, P. R. China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, P. R. China.|Department of Stomatology, Huashan Hospital, Fudan University, Shanghai, P. R. China.
authors.forename: Shangfeng
authors.initials: S
authors.lastname: Liu
mesh_terms: D000367:Age Factors; D000375:Aging / Q000378:metabolism / Q000473:pathology; D017209:Apoptosis / Q000187:drug effects; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects*; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair / Q000187:drug effects; D003795:Dental Sac / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation; D006801:Humans; D006918:Hydroxyurea / Q000633:toxicity*; D008875:Middle Aged; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D017382:Reactive Oxygen Species / Q000378:metabolism; D013234:Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D055815:Young Adult; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; D001616:beta-Galactosidase; D006918:Hydroxyurea
keywords: DNA damage; Gerotarget; aging; cellular senescence model; dental stem cells; stress