Dong-Jun Kim; Seong-Hwan Moon; Hyang Kim; Eun-Hae Kwon; Soo-Bong Hahn; Yong-Min Cheon; Hak-Sun Kim; Kyeong-Jin Han; Moon-Soo Park; Hwan-Mo Lee

doi:10.4184/jkss.2002.9.3.165

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Kim, Moon, Kim, Kwon, Hahn, Cheon, Kim, Han, Park, and Lee: Transforming Growth Factor-β1 and Bone Morphogenetic Protein-2 Upregulates Matrix Synthesis and Chondrogenic Phenotype in Intervertebral Disc Cells

Original Article

Journal of Korean Spine Surg 2002;9(3):165-171.

Published online: 21 February 2002

DOI: https://doi.org/10.4184/jkss.2002.9.3.165

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., Hwan-Mo Lee, M.D.

Department of Orthopaedic Surgery, Yonsei University, Seoul, Korea

^∗Ewha Women, College of Medicine, Seoul, Korea

^∗∗University, 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, E-mail : hwanlee@yumc.yonsei.ac.kr

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Objectives

To determine effect of transforming growth factor- β 1 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- β 1 was purchased from R&D and BMP- 2 was produced by transfection of pcDNA 3.1/Hygro/BMP- 2 to CHO cell using Lipofectamine 2000. rhBMP- 2 was separated by Heparin- Sepharose A chromatography. TGF- β 1 and 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- β 1 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- β 1 and BMP- 2 proved to be effective anabolic agent for maximizing matrix synthesis without evidence of osteogenesis.

Keywords: TGF- β1, BMP- 2, A ggrecan, Collagen I, Collagen II, Osteocalcin

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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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