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아텔로콜라겐 지지체와 성장인자에 의한 조직 공학적 추간판의 재생

Other Titles
 Tissue Engineered Intervertebral Disc by Atelocollagen Scaffolds and Growth Factors 
 이광일  ;  문성환  ;  김정환  ;  박시내  ;  서활  ;  최윤진  ;  박시영  ;  김학선  ;  이환모 
 Journal of Korean Society of Spine Surgery (대한척추외과학회지), Vol.12(3) : 164-173, 2005 
Journal Title
Journal of Korean Society of Spine Surgery(대한척추외과학회지)
Issue Date
Intervertebral disc ; Atelocollagen ; TGF-β1 ; BMP-2 ; Tissue engineering
Intervertebral disc ; Atelocollagen ; TGF-β1 ; BMP-2 ; Tissue engineering
Study Design: In vitro experimental study.

Objectives: To examine the cellular proliferation, synthetic activity and phenotypical expression of intervertebral disc (IVD) cells seeded on types I and II atelocollagen scaffolds, with the stimulation of TGF-β1 and BMP-2.
Summary of Literature Review: Recently, tissue engineering is regarded as a new experimental technique for the biological treatment of degenerative IVD diseases, and has been highlighted as a promising technique for the regeneration of tissues and organs in the human body. Research on cell transplantation in artificial scaffolds has provided that the conditions for tissue engineering have to be equilibrated, including the cell viability and proliferation, maintenance of characteristic phenotype, suitable scaffolds in organisms and biologically stimulated growth factor.

Material and Method: Lumbar IVD cells were harvested from 10 New Zealand white rabbits, with the nucleus pulposus cells isolated by sequential enzymatic digestion. Each of 1% types I and II atelocollagen dispersions were poured into a 96-well plate (diameter 5 mm), frozen at -70℃, and then lyophilized at -50℃. Fabricated porous collagen matrices were made using the cross-linking method. Cell suspensions were imbibed by surface tension into a scaffold consisting of atelocollagen. The cell cultured scaffolds were then treated with TGF-β1 (10 ng/ml) or BMP-2 (100 ng/ml) or both. After 1 and 2 week culture periods, the DNA synthesis was measured by [3H] thymidine incorporation, and newly synthesized proteoglycan by incorporation of [35S] sulphate. Reverse transcription-polymerase chain reactions for the mRNA expressions of type I and II collagen, aggrecan and osteocalcin were performed. The inner morphology of the scaffolds was determined by scanning electron microscopy (SEM).

Results: The IVD cultures in collagen type II with TGF-β1 demonstrated an increase in proteoglycan synthesis and up regulation of aggrecan and types I and II collagen mRNA expressions compared to the control. IVD cultures in the type I atelocollagen scaffold with growth factors exhibited an increase in DNA synthesis and up regulation of the type II atelocollagen mRNA expression. With all combinations of growth factor, the IVD cultures in types I and II atelocollagen scaffolds showed no up regulation of the osteocalcin mRNA expression. Furthermore, there was no synergistic effect of TGF-β1 and BMP-2 in the matrix synthesis or for the mRNA expression of the matrix components.

Conclusions: Nucleus pulposus cells from rabbit were viable in atelocollagen types I and II atelocollagen scaffolds. The type I atelocollagen scaffold was suitable for cell proliferation, but the type II atelocollagen scaffold was more suitable for extracellular matrix synthesis. The IVD cells in both scaffolds were biologically responsive to growth factors. Taken together, nucleus pulposus cells in atelocollagen scaffolds, with anabolic growth factors, provide a mechanism for tissue engineering of IVD cells.
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Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Medical Engineering (의학공학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Orthopedic Surgery (정형외과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Yonsei Biomedical Research Center (연세의생명연구원) > 1. Journal Papers
Yonsei Authors
Kim, Hak Sun(김학선) ORCID logo https://orcid.org/0000-0002-8330-4688
Moon, Seong Hwan(문성환)
Park, Si Nae(박시내)
Park, Si Young(박시영)
Suh, Hwal(서활)
Lee, Kwang Il(이광일)
Lee, Hwan Mo(이환모) ORCID logo https://orcid.org/0000-0002-5405-3832
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