title: Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells.
abstract: OBJECTIVE
  To construct a recombinant adenovirus vector-carrying human growth and differentiation factor-5 (GDF-5) gene, investigate the biological effects of adenovirus-mediated GDF-5 (Ad-GDF-5) on extracellular matrix (ECM) expression in human degenerative disc nucleus pulposus (NP) cells, and explore a candidate gene therapy method for intervertebral disc degeneration (IDD).
  
  
  METHODS
  Human NP cells of a degenerative disc were isolated, cultured, and infected with Ad-GDF-5 using the AdEasy-1 adenovirus vector system. On Days 3, 7, 14, and 21, the contents of the sulfated glycosaminoglycan (sGAG), deoxyribonucleic acid (DNA) and hydroxyproline (Hyp), synthesis of proteoglycan and collagen II, gene expression of collagen II and aggrecan, and NP cell proliferation were assessed.
  
  
  RESULTS
  The adenovirus was an effective vehicle for gene delivery with prolonged expression of GDF-5. Biochemical analysis revealed increased sGAG and Hyp contents in human NP cells infected by Ad-GDF-5 whereas there was no conspicuous change in basal medium (BM) or Ad-green fluorescent protein (GFP) groups. Only cells in the Ad-GDF-5 group promoted the production of ECM, as demonstrated by the secretion of proteoglycan and up-regulation of collagen II and aggrecan at both protein and mRNA levels. The NP cell proliferation was significantly promoted.
  
  
  CONCLUSIONS
  The data suggest that Ad-GDF-5 gene therapy is a potential treatment for IDD, which restores the functions of degenerative intervertebral disc through enhancing the ECM production of human NP cells.
authors.affiliation: Research Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital and the Second Clinical Institute of North Sichuan Medical College, Nanchong 637000, China.
authors.forename: Xu-wei
authors.initials: XW
authors.lastname: Luo
authors.affiliation: Research Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital and the Second Clinical Institute of North Sichuan Medical College, Nanchong 637000, China.
authors.forename: Kang
authors.initials: K
authors.lastname: Liu
authors.affiliation: Research Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital and the Second Clinical Institute of North Sichuan Medical College, Nanchong 637000, China.
authors.forename: Zhu
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Research Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital and the Second Clinical Institute of North Sichuan Medical College, Nanchong 637000, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Research Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital and the Second Clinical Institute of North Sichuan Medical College, Nanchong 637000, China.
authors.forename: Xiao-wei
authors.initials: XW
authors.lastname: Han
authors.affiliation: Research Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital and the Second Clinical Institute of North Sichuan Medical College, Nanchong 637000, China.
authors.forename: Yi-guang
authors.initials: YG
authors.lastname: Bai
authors.affiliation: Research Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital and the Second Clinical Institute of North Sichuan Medical College, Nanchong 637000, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Feng
mesh_terms: D000256:Adenoviridae / Q000502:physiology*; D002460:Cell Line; D005109:Extracellular Matrix / Q000378:metabolism* / Q000648:ultrastructure; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D055428:Growth Differentiation Factor 5 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007403:Intervertebral Disc / Q000254:growth & development* / Q000648:ultrastructure; D055959:Intervertebral Disc Degeneration / Q000378:metabolism* / Q000473:pathology; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D016326:Extracellular Matrix Proteins; D055428:Growth Differentiation Factor 5
keywords: Adenovirus; Degeneration; Gene therapy; Growth and differentiation factor-5 (GDF-5); Intervertebral disc; Nucleus pulposus