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Biotech Histochem. 2009 Dec;84(6):287-94. doi: 10.3109/10520290903116884.

Exogenous thymosin beta4 prevents apoptosis in human intervertebral annulus cells in vitro.

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  • 1Department of Orthopaedic Surgery, Carolinas Medical Center, Charlotte, North Carolina 28232, USA.

Abstract

Loss of cells in the human disc due to programmed cell death (apoptosis) is a major factor in the aging and degenerating human intervertebral disc. Our objective here was to determine if thymosin beta(4) (TB4), a small, multifunctional 5 kDa protein with diverse activities, might block apoptosis in human annulus cells cultured in monolayer or three-dimensional (3D) culture. Apoptosis was induced in vitro using hydrogen peroxide or serum starvation. Annulus cells were processed for identification of apoptotic cells using the TUNEL method. The percentage of apoptotic cells was determined by cell counts. Annulus cells also were treated with TB4 for determination of proliferation, and proteoglycan production was assessed using cell titer and 1,2 dimethylmethylamine (DMB) assays and histological staining. A significant reduction in disc cell apoptosis occurred after TB4 treatment. The percentage of cells undergoing apoptosis decreased significantly in TB4 treated cells in both apoptosis induction designs. TB4 exposure did not alter proteoglycan production as assessed by either DMB measurement or histological staining. Our results indicate the need for further studies of the anti-apoptotic effect of TB4 and suggest that TB4 may have therapeutic application in future biological therapies for disc degeneration.

PMID:
20055734
PMCID:
PMC2886502
DOI:
10.3109/10520290903116884
[PubMed - indexed for MEDLINE]
Free PMC Article
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