Open Access, Peer-reviewed
pISSN 2093-4378
eISSN 2093-4386
    J Korean Soc Spine Surg. 2002 Sep;9(3):165-171. Korean.
    Published online Sep 30, 2002.
    https://doi.org/10.4184/jkss.2002.9.3.165
    Copyright © 2002 Korean Society of Spine Surgery
    Original Article

    Transforming Growth Factor-β1 and Bone Morphogenetic Protein-2 Upregulates Matrix Synthesis and Chondrogenic Phenotype in Intervertebral Disc Cells

    Dong-Jun Kim, M.D.,* Seong-Hwan Moon, M.D., Hyang Kim, M.S., Eun-Hae Kwon, B.S., Soo-Bong Hahn, M.D., Yong-Min Cheon, M.D., Hak-Sun Kim, M.D., Kyeong-Jin Han, M.D.,** Moon-Soo Park, M.D. and Hwan-Mo Lee, M.D.
      • Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
      • *Department of Orthopaedic Surgery, Ewha Women University College of Medicine, Seoul, Korea.
      • **Department of Orthopaedic Surgery, Aju University College of Medicine, Seoul, Korea.
    • Address reprint requests to Hwan-Mo Lee, M.D. Department of Orthopaedic Surgery, College of Medicine, Yonsei University, #134 Shinchon-Dong, Soedaemun-ku, Seoul 120-752, Korea. Tel: 82-2-361-5649, Fax: 82-2-363-1139, Email: hwanlee@yumc.yonsei.ac.kr

    Abstract

    Objectives

    To determine effect of transforming growth factor-β1 and bone morphogenetic protein-2 in matrix synthesis and expression of chondrogenic phenotype in human intervertebral disc cells.

    Materials and Methods

    The intervertebral disc cells were harvested and cultured from the surgical patients for the degenerative disc disease. TGF-β1 was purchased from R&D and BMP-2 was produced by transfection of pcDNA3.1/Hygro/BMP-2 to CHO cell using Lipofectamine 2000. rhBMP-2 was separated by Heparin-Sepharose A chromatography. TGF-β1 a n d BMP-2 were administered to culture. Proteoglycan synthesis was assessed by 35S incorporation and expression of matrix mRNA was analyzed by RT-PCR for collagen I, collagen II, aggrecan, and osteocalcin.

    Results

    TGF-β1 and BMP-2 showed increased proteoglycan synthesis and expression of collagen I, collagen II and aggrecan mRNA in dose dependent manner respectively. There was no recognizable synergistic effect in matrix synthesis and matrix mRNA expression. Throughout dosage, expression of osteogenic phenotype (osteocalcin mRNA) was not noted.

    Conclusion

    TGF-β1 and BMP-2 proved to be effective anabolic agent for maximizing matrix synthesis without evidence of osteogenesis.

    Keywords
    TGF-β1; BMP-2; Aggrecan; Collagen I; Collagen II; Osteocalcin

    Figures

    Fig. 1
    Newly synthesized proteoglycan as measured by incorporation 35S-sulfate according to the dose of TGF-b1 and BMP-2. In given doses, proteoglycan synthesis was upregulated compared to control culture (p<0.05).

    Fig. 2
    Dose response of BMP-2 in mRNA expression of collagen type I, collagen type II, aggrecan, and osteoclacin as measured by reverse transcriptase-polymerase chain reaction. Collagen type I, collagen type II, and aggrecan mRNA expression were upregulated, while osteoclacin mRNA showed no recognizable upregulation.

    Fig. 3
    Effect of BMP-2 and TGF-b1 in mRNA expression of collagen type I, collagen type II, aggrecan, and osteoclacin as measured by reverse transcriptase-polymerase chain reaction. Even with combination of two growth factors, there was no synergistic effect in matrix mRNA expression. In given doses, there was no expression of osteocalcin mRNA.

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    MeSH Terms
    Aggrecans
    Animals
    CHO Cells
    Chromatography
    Collagen
    Cricetinae
    Humans
    Intervertebral Disc*
    Osteocalcin
    Osteogenesis
    Phenotype*
    Proteoglycans
    RNA, Messenger
    Transfection
    Transforming Growth Factor beta1
    Metrics
    Share
    Figures
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