title: IGF-1 rescues human intervertebral annulus cells from in vitro stress-induced premature senescence.
abstract: The aging human intervertebral disc contains a sizeable population of senescent cells. Since senescent cells cannot divide, senescence reduces the disc's ability to generate new cells to replace existing ones lost to necrosis or apoptosis. The objectives of the present work were: (1) to develop a reliable in vitro model for stress-induced premature senescence in human annulus cells, and (2) to investigate the potential for insulin-like growth factor-1 (IGF-1) to prevent or ameliorate senescence in vitro. The developed experimental model employs a 2 h exposure to 50 microM hydrogen peroxide; immunocytochemical localization of senescence associated-beta-galactosidase at pH 6.0 was used as the marker for senescent cells, and the percentage of senescent cells quantified after 3 days of culture. Nine sets of annulus cells were obtained from eight human surgical disc specimens; cells were tested with 0, 50, 100 or 500 ng/ml IGF-1. Although 50 or 100 ng/ml IGF-1 did not significantly alter the percentage of senescent cells, a significant reduction was present following exposure to 500 ng/ml IGF-1 (control, 56.3% +/- 8.5 (9); mean +/- SEM, (n) vs. treated, 39.6% +/- 6.6 (9), p = 0.0009). These novel findings point to the value of continued research towards development of future biologic therapies designed to reduce cell senescence in degenerating human discs.
authors.affiliation: Department of Orthopaedic Surgery, Carolinas Medical Center, PO Box 32861, Charlotte, NC 28232, USA. helen.gruber@carolinashealthcare.org
authors.forename: Helen E
authors.initials: HE
authors.lastname: Gruber
authors.forename: Gretchen L
authors.initials: GL
authors.lastname: Hoelscher
authors.forename: Jane A
authors.initials: JA
authors.lastname: Ingram
authors.forename: Synthia
authors.initials: S
authors.lastname: Bethea
authors.forename: Edward N
authors.initials: EN
authors.lastname: Hanley
mesh_terms: D018929:Cell Culture Techniques; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007150:Immunohistochemistry; D007334:Insulin-Like Growth Factor I / Q000378:metabolism / Q000494:pharmacology*; D007403:Intervertebral Disc / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects*; D013122:Spinal Diseases / Q000473:pathology*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D016877:Oxidants; D007334:Insulin-Like Growth Factor I; D006861:Hydrogen Peroxide; D001616:beta-Galactosidase