For gene expression information, ratio values had been log transformed prior to evaluation by a two way ANOVA plus a post hoc Bonferroni corrected t test. In all cases, a amount of 5% was viewed as statistically substantial, Benefits FN f maximally improved catabolic activities at 5% oxygen tension The capacity of FN f to influence the levels of NO, PGE2 and GAG synthesis in an oxygen dependent manner have been when compared with IL 1B in constructs cultured for 48 hours, At 1, five and 21% oxygen tension, NO production was enhanced with FN f or IL 1B in comparison with untreated controls, At 5% oxygen tension, FN f elevated NO production with values signifi cantly greater than cytokine treated constructs, At all oxygen tensions, co incubation with L NIO lowered fragment or cytokine induced NO levels with values returning to basal levels, PGE2 release was increased with either FN f or IL 1B at 5 and 21% oxygen tension but not at 1% oxygen tension, Co incubation with L NIO partially decreased PGE2 levels in FN f treated constructs cultured at 5 and 21% oxygen tension.
In addition, FN f reduced GAG synthesis at 1, 5 and 21% oxygen tension, In contrast, selelck kinase inhibitor the cytokine decreased GAG syn thesis at 1% oxygen tension but not at five or 21% oxygen tension. At 1 or 5% oxygen tension, frag ment or cytokine induced inhibition on GAG synthesis was reversed with L NIO, Compression abolished FN f induced catabolic effects at five and 21% oxygen tension Because the catabolic effects in response to FN f had been maximal at 5% oxygen tension, additional ex vivo research ex amined the effect of dynamic compression in constructs cultured at five and 21% oxygen tension and compared the response to IL 1B.
Figure 4 reveals that dynamic compres sion inhibits NO release at 5 and 21% oxygen tension, In unstrained constructs, FN f en hanced NO production with values highest at 5% oxygen tension when compared to the effects inhibitor EGFR Inhibitor of IL 1B, The enhanced production of NO by fragment, cytokine or low oxygen tension was decreased with dy namic compression, Stimula tion with compression as well as the NOS inhibitor lowered NO levels further in fragment or cytokine treated con structs, At 5% oxygen tension, dynamic compression inhibits PGE2 release in untreated constructs, In unstrained constructs, the presence from the fragment or cytokine enhanced PGE2 release with values broadly comparable and ranging from 7. six to 8. 0 pg ml for constructs cultured at 5 or 21% oxygen tension. Stimulation with dynamic compression and or L NIO reduced PGE2 levels to basal values in an oxygen independent manner with all the magnitude in inhibition ranging between 77 and 87%. In untreated samples, total MMP activity was inhibited with dynamic compression at 5% oxygen tension.
The fragment or cytokine elevated MMP activity with max imal values at 5% oxygen tension for constructs cultured with FN f when in comparison to IL 1B, At 5 or 21% oxygen tension, stimulation with dynamic compres sion or the NOS inhibitor abolished fragment or cyto kine induced MMP activity, Moreover, dynamic compression enhanced GAG synthesis using a higher magnitude in stimu lation for constructs cultured at 21% oxygen tension when when compared with 5%, At five and 21% oxy gen tension, the presence on the FN f or IL 1B lowered GAG synthesis, This response was reversed by stimulation with dynamic compression and L NIO at 5 and 21% oxygen tension.