title: [Effect of adenovirus human bone morphogenetic protein 4 on human degenerative lumbar intervertebral disc cells].
abstract: OBJECTIVE
  To research the transfer of adenovirus human bone morphogenetic protein 4 (Ad-hBMP-4) to human degenerative lumbar intervertebral disc cells in vitro and analyze its effect on the proteoglycan, collagen type II, and Sox9 of intervertebral disc cells.
  
  
  METHODS
  Identified Ad-hBMP-4 was amplified and detected. Degenerative lumbar intervertebral disc cells were aspirated from the degenerative lumbar intervertebral disc of patients with Modic III level disc protrusion (aged, 27-50 years). The expressing position of collagen type II was identified in the intervertebral disc cells through the laser confocal microscope. The intervertebral disc cells at passage 1 were transfected with Ad-hBMP-4 as experimental group. After 3 and 6 days of transfection, RT-PCR was used to detect the mRNA expressions of proteoglycan, collagen type II, and Sox9, and Western blot to detect the expressions of proteoglycan and collagen type II proteins. Non-transfected cells at passage 1 served as control group.
  
  
  RESULTS
  The virus titer of Ad-hBMP-4 was 5 x 10(6) PFU/mL. No morphological changes in the cells after transfection by Ad-hBMP-4. Collagen type II mainly expressed in the cell cytoplasm. The mRNA expressions of the proteoglycan, collagen type II, and Sox9 in experimental group at 3 and 6 days after transfection were significantly higher than those in control group by RT-PCR (P < 0.05), and the expressions of proteoglycan and collagen type II proteins were significantly higher than those in contorl group by Western blot (P < 0.05). There were significant differences between 3 days and 6 days in experimental group (P < 0.05).
  
  
  CONCLUSIONS
  Ad-hBMP-4 could transfect human degenerative lumbar intervertebral cells with high efficiency and promote collagen type II, proteoglycan, and Sox9 expressions. hBMP-4 may play an important role in the repair process during early disc degeneration.
authors: Department of Orthopaedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, P.R.China.
authors: Dejun
authors: D
authors: Cao
authors: Zhengxue
authors: Z
authors: Quan
authors: Dianming
authors: D
authors: Jiang
authors: Xiaoji
authors: X
authors: Luo
authors: Weiyang
authors: W
authors: Zhong
authors: Gang
authors: G
authors: Liu
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000328:Adult; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology / Q000378:metabolism*; D008159:Lumbar Vertebrae; D008875:Middle Aged; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D024043:Collagen Type II; D011509:Proteoglycans; D012333:RNA, Messenger; D011994:Recombinant Proteins; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human