title: Biologic response of degenerative living human nucleus pulposus cells to treatment with cytokines.
abstract: OBJECTIVE
  To investigate the molecular responses of various genes and proteins related to disc degeneration upon treatment with cytokines that affect disc-cell proliferation and phenotype in living human intervertebral discs (IVDs). Responsiveness to these cytokines according to the degree of disc degeneration was also evaluated.
  
  
  METHODS
  The disc specimens were classified into two groups: group 1 (6 patients) showed mild degeneration of IVDs and group 2 (6 patients) exhibited severe degeneration of IVDs. Gene expression was analyzed after treatment with four cytokines: recombinant human bone morphogenic protein (rhBMP-2), transforming growth factor-β (TGF-β), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α). Molecular responses were assessed after exposure of cells from the IVD specimens to these cytokines via real-time polymerase chain reaction and immunofluorescence staining.
  
  
  RESULTS
  mRNA gene expression was significantly greater for aggrecan, type I collagen, type II collagen, alkaline phosphatase, osteocalcin, and Sox9 in group 1 than mRNA gene expression in group 2, when the samples were not treated with cytokines. Analysis of mRNA levels for these molecules after morphogen treatment revealed significant increases in both groups, which were much higher in group 1 than in group 2. The average number of IVD cells that were immunofluorescence stained positive for alkaline phosphatase increased after treatment with rhBMP-2 and TGF-β in group 1.
  
  
  CONCLUSIONS
  The biologic responsiveness to treatment of rhBMP-2, TGF-β, TNF-α, and IL-1β in the degenerative living human IVD can be different according to the degree of degeneration of the IVD.
authors.affiliation: Department of Neurosurgery, Ajou University College of Medicine, Suwon, Korea.
authors.forename: Sang Hyun
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Department of Neurosurgery, The Spine and Spinal Cord Institute, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea. kuhsu@yuhs.ac.
authors.forename: Sung Uk
authors.initials: SU
authors.lastname: Kuh
authors.affiliation: Department of Neurosurgery, The Spine and Spinal Cord Institute, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Keung Nyun
authors.initials: KN
authors.lastname: Kim
authors.affiliation: Department of Neurosurgery, The Spine and Spinal Cord Institute, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Jeong Yoon
authors.initials: JY
authors.lastname: Park
authors.affiliation: Department of Neurosurgery, Ajou University College of Medicine, Suwon, Korea.
authors.forename: Ki Hong
authors.initials: KH
authors.lastname: Cho
authors.affiliation: Department of Neurosurgery, The Spine and Spinal Cord Institute, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Dong Kyu
authors.initials: DK
authors.lastname: Chin
authors.affiliation: Department of Neurosurgery, The Spine and Spinal Cord Institute, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Keun Su
authors.initials: KS
authors.lastname: Kim
authors.affiliation: Department of Neurosurgery, The Spine and Spinal Cord Institute, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Yong Eun
authors.initials: YE
authors.lastname: Cho
mesh_terms: D000328:Adult; D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D001688:Biological Products / Q000494:pharmacology / Q000627:therapeutic use*; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology / Q000627:therapeutic use; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000494:pharmacology* / Q000627:therapeutic use*; D005260:Female; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology / Q000627:therapeutic use; D007403:Intervertebral Disc / Q000187:drug effects* / Q000473:pathology*; D055959:Intervertebral Disc Degeneration / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology / Q000627:therapeutic use; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology / Q000627:therapeutic use; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D053674:Aggrecans; D001688:Biological Products; D055396:Bone Morphogenetic Protein 2; D024042:Collagen Type I; D024043:Collagen Type II; D016207:Cytokines; D007375:Interleukin-1; D012333:RNA, Messenger; D011994:Recombinant Proteins; D055758:SOX9 Transcription Factor; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; C499469:recombinant human bone morphogenetic protein-2; D015675:Osteocalcin; D000469:Alkaline Phosphatase
keywords: Cytokine; IL-1β; TGF-β; TNF-α; disc degeneration; rhBMP-2