Bone morphogenetic protein-7 protects human intervertebral disc cells in vitro from apoptosis

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Abstract
Background context: Disc degeneration includes dysfunction and loss of disc cells leading to a decrease in extracellular matrix (ECM) components. Apoptosis has been identified in degenerated discs. Bone morphogenetic protein-7 (BMP-7) has been reported to stimulate ECM synthesis in the intervertebral disc (IVD), but its effect on disc cell viability is unknown. Purpose: To investigate whether BMP-7 can protect disc cells from programmed cell death while enhancing ECM production. Study design: An in vitro study to examine the effect of BMP-7 on apoptosis of IVD cells. Methods: Human nucleus pulposus (NP) cells were cultured in monolayer, and human recombinant pure BMP-7 (rhBMP-7) was added to the medium when the cells were in the second passage. Thereafter, apoptosis was induced by either tumor necrosis factor-alpha (TNF-α) or hydrogen peroxide (H2O2). Cellular apoptosis was evaluated by terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling assay and caspase-3 activity. ECM synthesis was assessed by immunofluorescence for collagen-2 and aggrecan. To study the possibility of bone induction by rhBMP-7 in disc cells, alkaline phosphatase activity and Alizarin red-S staining were evaluated. Results: Apoptosis was induced by both TNF-α and H2O2. Addition of rhBMP-7 resulted in inhibition of the apoptotic effects caused by both inducers. Further, BMP-7 decreased caspase-3 activity. In the presence of BMP-7, ECM production was maintained by the cells despite being in an apoptotic environment. No osteoblastic induction of the disc cells was seen. Conclusions: BMP-7 was demonstrated to prevent apoptosis of human disc cells in vitro. One of the antiapoptotic effects of BMP-7 on NP cells might be a result of its inactivation of caspase-3. Collagen production was maintained by addition of rhBMP-7 in an apoptotic environment.
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Author(s)
Wei, Ai-Qun
Brisby, Helena
Chung, Sylvia
Diwan, Ashish
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Publication Year
2008
Resource Type
Journal Article
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UNSW Faculty