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Updated: Jun 21, 2026
Methods for the Modulation and Analysis of NF-κB-dependent Adult Neurogenesis
Published on: February 13, 2014
Dynamic compression alters NFkappaB activation and IkappaB-alpha expression in IL-1beta-stimulated
O O Akanji1, P Sakthithasan, D M Salter
1Queen Mary University of London, UK.
Objective And Design:
Determine the effect of IL-1beta and dynamic compression on NFkappaB activation and IkappaB-alpha gene expression in chondrocyte/agarose constructs.
Methods:
Constructs were cultured under free-swelling conditions or subjected to dynamic compression for up to 360 min with IL-1beta and/or PDTC (inhibits NFkappaB activation). Nuclear translocation of NFkappaB-p65 was analysed by immunofluoresence microscopy. Gene expression of IkappaB-alpha, iNOS, IL-1beta and IL-4 was assessed by real-time qPCR.
Results:
Nuclear translocation of NFkappaB-p65 was concomitant with an increase in nuclear fluorescence intensity which reached maximum values at 60 min with IL-1beta (p < 0.001). Dynamic compression or PDTC reduced nuclear fluorescence and NFkappaB nuclear translocation in cytokine-treated constructs (p < 0.001 and p < 0.01 respectively). IL-1beta increased IkappaB-alpha expression (p < 0.001) at 60 min and either induced iNOS (p < 0.001) and IL-1beta (p < 0.01) or inhibited IL-4 (p < 0.05) expression at 360 min. These time-dependent events were partially reversed by dynamic compression or PDTC (p < 0.01) with IL-1beta. Co-stimulation by dynamic compression and PDTC favoured suppression (IkappaB-alpha, iNOS, IL-1beta) or induction (IL-4) of gene expression.
Conclusions:
NFkappaB is one of the key players in the mechanical and inflammatory pathways, and its inhibition by a biophysical/therapeutic approach could be a strategy for attenuating the catabolic response in osteoarthritis.
