Pre-Clinical Study on Natural Eggshell Membrane Evaluating Possible Mode of Action
ABSTRACT: Tumor necrosis factor alpha (TNF-a) plays an important role in inflammatory processes. This study examined the effects of NEM® (Natural Eggshell Membrane) on IL-2, IL-4, IL-6, IL-10, interferon gamma (IFN-g) and TNF-a cytokine production by 4-day peripheral blood mononuclear cell (PBMC) cultures exposed to serial dilutions of either an aqueous extract of NEM® (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-g and TNF-a cytokine production. NEM-AQ reduced levels of IL-6, IL-10, IFN-g 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. The in vitro digest of NEM® (NEM-IVD), at the two lowest concentrations of product, significantly reduced TNF-a production by PBMC cultures exposed to PWM compared to 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 cell-specific mitogen PHA, as well as the mitogen PWM that requires cellular cross-talk and these effects may be partially mediated through a reduction in the pro-inflammatory cytokine TNF-a. The suppression of TNF-a production in the presence of the in vitro digested NEM-IVD is promising for NEM® as a consumable anti-inflammatory product.
Full published study: Benson, K.F., Ruff, K.J., and Jensen, G.S. 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. Journal of Medicinal Food, (2012) 15(4):360-368.
Safety Evaluation of a Natural Eggshell Membrane-derived Product
ABSTRACT: Natural Eggshell Membrane is a dietary supplement ingredient that contains naturally occurring glycosaminoglycans and proteins essential for maintaining healthy joint and connective tissues. Natural Eggshell Membrane was evaluated for safety via in vitro and in vivo toxicological studies. This included testing for cytotoxicity, genotoxicity, acute oral toxicity, and 90-day repeated-dose oral toxicity. Natural Eggshell Membrane did not exhibit any cytotoxic effects at a dose of 100 mg in an in vitro human cell viability assay after incubation for up to 20 hours. Natural Eggshell Membrane did not exhibit any genotoxic effects in an in vitro assay of four strains of histidine-dependent Salmonella typhimurium and one strain of tryptophan-dependent Escherichia coli at a dose of up to 5,000 mg/plate. Natural Eggshell Membrane did not exhibit any signs of acute toxicity in rats at a single oral dose of up to 2,000 mg/kg body weight, nor signs of toxicity (via urinalysis, hematology, clinical chemistry, or histopathological evaluation) in rats at a repeated oral dose of up to 2,000 mg/kg body weight per day for 90 days. The results of these studies suggest that Natural Eggshell Membrane may be safe for human consumption.
Full published study: Safety Evaluation of a Natural Eggshell Membrane-derived Product. Kevin J. Ruff, Ph.D.a, John R. Endres, N.D.b*, Amy E. Clewell, N.D.c, James R. Szabo, D.V.M, Ph.D., DACVPd, Alex G. Schauss, Ph.D., FACNe. Food and Chemical Toxicology (2012).
Effects of eggshell membrane on pain and stiffness of the knee: a randomized, multicenter, double-blind, placebo-controlled clinical study.
ABSTRACT: Natural Eggshell Membrane, manufactured by ESM Technologies, is a dietary supplement ingredient that contains naturally occurring glycosaminoglycans and proteins essential for maintaining healthy articular cartilage and the surrounding synovium. The randomized, multicenter, double-blind, placebo-controlled clinical study was conducted to evaluate the efficacy and safety of eggshell membrane as a treatment for pain and stiffness associated with the knee. Sixty-seven patients were randomly assigned to receive either oral eggshell membrane 500 mg (n=34) or placebo (n=33) daily for 8 weeks. The primary endpoint was the change in overall Western Ontario and McMasters Universities (WOMAC) Osteoarthritis Index as well as pain, stiffness, and function WOMAC subscales measured at 10, 30, and 60 days. The clinical assessment was performed on the intent-to-treat population. Supplementation with eggshell membrane produced an absolute rate of response that was statistically significant (up to 26.6%) versus placebo at all time points for both pain and stiffness, but was not significantly improved for function and overall WOMAC scores, although trending toward improvement. Rapid responses were seen for mean pain subscores (15.9% reduction, P=0.036) and mean stiffness subscores (12.8% reduction, P=0.024) occurring after only 10 days of supplementation. There were no serious adverse events reported during the study and the treatment was reported to be well tolerated by study participants. Natural Eggshell Membrane is shown to be an effective and safe option for the treatment of pain and stiffness associated with the knee. Supplementation with eggshell membrane, 500 mg taken once daily, significantly reduced both joint pain and stiffness compared to placebo at 10, 30, and 60 days. The Clinical Trial Registration number for this study is NCT00750477.
Full published study: Kevin J. Ruff, Anne Winkler, Robert W. Jackson, Dale P. DeVore, Barry W. Ritz. Eggshell membrane in the treatment of pain and stiffness from osteoarthritis of the knee: a randomized, multicenter, double-blind, placebo-controlled clinical study. Clinical Rheumatology, April 2009: DOI 10.1007/s10067-009-1173-4.
Eggshell membrane: A possible new natural therapeutic for joint and connective tissue. Results from two open-label human clinical studies.
ABSTRACT: Natural Eggshell Membrane, manufactured by ESM Technologies, is a possible new effective and safe therapeutic option for the treatment of pain and inflexibility associated with joint and connective tissue (JCT) disorders. Supplementation with eggshell membrane, 500 mg taken once daily, significantly reduced pain, both rapidly (seven days) and continuously (30 days). It also showed clinically meaningful results from a brief responder analysis, demonstrating that significant proportions of treated patients may be helped considerably from eggshell membrane supplementation. The Clinical Trial Registration numbers for these trials are: NCT00750230 and NCT00750854.
Full published study: Kevin J Ruff, Dale P DeVore, Michael D Leu, Mark A Robinson. Eggshell membrane: A possible new natural therapeutic for joint and connective tissue disorders. Results from two open-label human clinical studies. Clinical Interventions in Aging 2009: 4 235–240.