title: Increased cell senescence is associated with decreased cell proliferation in vivo in the degenerating human annulus.
abstract: BACKGROUND
  During disc degeneration, there is a well-recognized loss of cells. This puts the remaining cell population at high risk for any further decrease in cell function or cell numbers. Cell senescence has recently been shown to be present in the aging/degenerating human disc. Senescent cell are viable, metabolically active, persist, and accumulate over time, but cannot divide. Little is known about the relationship between renewal of the disc cell population via cell proliferation and disc cell senescence.
  
  
  OBJECTIVE
  To determine the percentage of senescent cells and proliferating cells in the human annulus in vivo.
  
  
  METHODS
  Human annulus specimens were obtained from surgical subjects and control donors in a study approved by the authors' Human Subjects Institutional Review Board.
  
  
  METHODS
  One Thompson Grade I disc, 4 Grade II discs, 9 Grade III discs, and 12 Grade IV discs were studied.
  
  
  METHODS
  The percentages of senescent cells and the percentage of proliferating cells.
  
  
  METHODS
  Immunohistochemistry was used to detect senescent cells using an antisenescence-associated beta-galactosidase antibody, and an antiproliferation antibody (Ki67). An average of 410 cells/specimens was counted to determine the percent senescence, and an average of 229 cells was counted to determine the percent proliferation.
  
  
  RESULTS
  Cell proliferation was low in both surgical and control normal donor annulus tissue (4.09%+1.77 (26), mean+SD (n)). There was no significant difference in the percentage of proliferating cells for more degenerate discs versus healthier discs (4.7%+1.6 (21) for Grades III and IV vs. 5.3%+1.9 (5) for Grades I and II). More degenerated Grades III and IV discs contained significantly greater percentages of senescent annulus cells than did the healthier Grades I and II discs (44.4%+20.0 (21) vs. 18.8%+11.0 (5), respectively; p=.011). A significant negative correlation was present between the percentage of senescent cells versus the percentage of proliferating cells, r=-0.013, p=.013. No correlation was present between age and the percentage of senescent cells or age and the percentage of proliferating cells.
  
  
  CONCLUSIONS
  Because senescent cells cannot divide, senescence may reduce the disc's ability to generate new cells to replace cells lost to necrosis or apoptosis. Senescent cells also accumulate in the disc over time, such that their metabolic patterns may contribute to the pathologic changes seen in degenerating discs. Novel data presented here show a significant negative correlation between the percentage of senescent cells and the percentage of proliferating cells during disc degeneration. Molecular work is underway in our lab to help us determine whether senescent cells in the disc secrete factors that can result in decreased proliferation in neighboring cells.
authors.affiliation: Department of Orthopaedic Surgery, Carolinas Medical Center, PO Box 32861, Cannon Research., Room 304, Charlotte, NC 28232, USA. helen.gruber@carolinashealthcare.org
authors.forename: Helen E
authors.initials: HE
authors.lastname: Gruber
authors.forename: Jane A
authors.initials: JA
authors.lastname: Ingram
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Davis
authors.forename: Edward N
authors.initials: EN
authors.lastname: Hanley
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D049109:Cell Proliferation; D016922:Cellular Senescence / Q000502:physiology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007403:Intervertebral Disc / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D013122:Spinal Diseases / Q000473:pathology*