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Tissue engineering of the intervertebral disc with cultured nucleus pulposus cells using atelocollagen scaffold and growth factors

 Lee, Kwang-Il  ;  Moon, Seong-Hwan  ;  Kim, Hyang  ;  Kwon, Un-Hye  ;  Kim, Ho-Joong  ;  Park, Si-Nae  ;  Suh, Hwal  ;  Lee, Hwan-Mo  ;  Kim, Hak-Sun  ;  Chun, Heoung-Jae  ;  Kwon, Il-Keun  ;  Jang, Ju-Woong 
 SPINE, Vol.37(6) : 1-7, 2012 
Journal Title
Issue Date
Animals ; Bone Morphogenetic Protein 2/pharmacology ; Cells, Cultured ; Collagen* ; Extracellular Matrix/drug effects ; Extracellular Matrix/metabolism ; Intercellular Signaling Peptides and Proteins/pharmacology ; Intervertebral Disc/cytology* ; Intervertebral Disc/drug effects ; Intervertebral Disc/metabolism ; Rabbits ; Tissue Engineering* ; Tissue Scaffolds* ; Transforming Growth Factor beta1/pharmacology
atelocollagen ; BMP-2 ; nucleus pulposus ; scaffold ; TGF- β1
STUDY DESIGN: In vitro experiment using rabbit nucleus pulposus (NP) cells seeded in atelocollagen scaffolds under the stimulation of growth factors. OBJECTIVE: To demonstrate the effect of anabolic growth factors in rabbit NP cells cultured in atelocollagen type I and type II. SUMMARY OF BACKGROUND DATA: Atelocollagen provides intervertebral disc (IVD) cells for a biocompatible environment to produce extracellular matrix. IVD cells with exogenous transforming growth factor-beta 1 (TGF-β1) and bone morphogenetic protein-2 (BMP-2) also render an increase in matrix synthesis. However, the effect of anabolic growth factors in NP cells cultured in atelocollagens was not elucidated before. METHODS: Rabbit NP cell was harvested, enzymatically digested, and cultured. The NP cells were seeded to atelocollagen type I and type II scaffolds, and then cultures were exposed to TGF-β1 (10 ng/mL) and/or BMP-2 (100 ng/mL). DNA synthesis was measured using [4H]-thymidine incorporation. Newly synthesized proteoglycan was measured using [35S]-sulfate incorporation. Reverse transcription-polymerase chain reactions (RT-PCRs) for mRNA expression of aggrecan, collagen type I, collagen type II, and osteocalcin were performed. RESULTS: Rabbit NP cells cultured in atelocollagen type I scaffold showed an increase (1.7 to 2.4-fold) in DNA synthesis in response to TGF-β1 and/or BMP-2 (P < 0.05), whereas NP cultures in atelocollagen type II demonstrated a 30% increase in DNA synthesis only with combination of both growth factors compared with control (P < 0.05). Rabbit NP cells in atelocollagen type II scaffold with TGF-β1 and combination of both growth factors exhibited robust 5.3- and 5.4-fold increases in proteoglycan synthesis (P < 0.05), whereas any cultures in atelocollagen type I failed to show any significant increase compared with control. Rabbit NP cells in atelocollagen type I and type II scaffolds with TGF-β1 and/or BMP-2 demonstrated the upregulation of aggrecan, collagen type I, and collagen type II mRNA expression compared with saline control (P < 0.05). The response in transcriptional level was more robust in atelocollagen type II than in type I. In any event, there is no recognizable expression of osteocalcin (P < 0.05). CONCLUSION: NP cells in atelocollagens under the stimulation of TGF-β1 and BMP-2 exhibited anabolic responses in transcriptional and translational levels. Hence, such an approach can provide a suitable engineered tissue for IVD regeneration with potential for robust refurbishment of matrix.
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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) > Medical Research Center (임상의학연구센터) > 1. Journal Papers
Yonsei Authors
Kwon, Un Hye(권언혜)
Kim, Hak Sun(김학선) ORCID logo https://orcid.org/0000-0002-8330-4688
Kim, Hyang(김향)
Kim, Ho Joong(김호중)
Moon, Seong Hwan(문성환)
Suh, Hwal(서활)
Lee, Kwang Il(이광일)
Lee, Hwan Mo(이환모) ORCID logo https://orcid.org/0000-0002-5405-3832
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