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Effects of Natural Eggshell Membrane (NEM) on Cytokine Production in Cultures of Peripheral Blood Mononuclear Cells: Increased Suppression of Tumor Necrosis Factor-a Levels After In Vitro Digestion

Kathleen F. Benson,1 Kevin J. Ruff,2 and Gitte S. Jensen

"ABSTRACT Tumor necrosis factor-a (TNF-a) plays an important role in inflammatory processes. This study examined the effects of natural eggshell membrane (NEM) (ESM Technologies, LLC, Carthage, MO, USA) on interleukin (IL)-2, IL-4, IL-6, IL-10, interferon-c (IFN-c), and TNF-a cytokine production by 4-day peripheral blood mononuclear cell (PBMC) cultures exposed to serial dilutions of either an aqueous extract of natural eggshell membrane (NEM-AQ) or NEM subjected to in vitro digestion (NEM-IVD). The effects on cytokine production were also assessed in the presence of phytohemagglutinin (PHA) and pokeweed mitogen (PWM) where exposure to NEM-AQ resulted in reduced levels of proliferation and statistically significant effects on IL-6, IL-10, IFN-c, and TNF-a cytokine production. NEM-AQ reduced levels of IL-6, IL-10, IFN-c, and TNF-a in cultures exposed to PHA. In cultures containing PWM, NEM-AQ reduced production of IL-10 and at the highest dose tested increased IL-6 and decreased TNF-a cytokine levels. NEM-IVD, at the two lowest concentrations of product, significantly reduced TNF-a production by PBMC cultures exposed to PWM compared with the in vitro digest control or native NEM. Taken together, these results suggest that NEM-AQ can influence signaling events in response to the T cellspecific mitogen PHA as well as to the mitogen PWM that require cellular cross-talk and that these effects may be partially mediated through a reduction in level of the pro-inflammatory cytokine TNF-a. The suppression of TNF-a production in the presence of NEM-IVD is promising for the use of NEM as a consumable anti-inflammatory product."

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