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Short-Term Effects of Ovomet®, Eggshell Membrane, in Joint Pain. A Double-Blind and Placebo Study
... There has been a growing trend toward companies that are actively involved in the industrial separation of eggshell membranes from waste eggshells. Eggnovo, based in Spain, uses calcium carbonate extracted from eggshells for applications in the food, food supplement, cosmetic, and pharmaceutical industries [13]. Just Egg, based in the United Kingdom, has developed a pilot plant for the production of eggshell powder in collaboration with the University of Leicester. ...
Direct methanol fuel cells (DMFCs) have great potential for use in portable electronics. However, obstacles such as methanol crossover, insufficient proton conductivity, and the high cost of Nafion hinder the broad commercialization of this technology. In line with the prevailing “waste-to-wealth” movement, eggshell powder was chosen as the filler for the Nafion matrix (rN-ES). Nano-calcium carbonate (nano-CaCO₃) was first produced from eggshell waste by a mechanochemical process before inclusion in the Nafion polymer matrix by the solution casting process. Cyclic voltammetry and electrochemical impedance spectroscopy were used to measure methanol permeability and proton conductivity. The composite membrane showed the highest value for ion exchange capacity of 1.25 mmol g⁻1 and water uptake of 46.54%. Remarkably, the through-plane method showed better proton conductivity (4.87 mS cm⁻1) compared to N117. The methanol permeability of the rN-ES composite membranes decreased to 3.3 times the permeability of N117. In the passive single-cell test of the DMFC, the use of a composite membrane with 5 wt.% nano-CaCO₃ resulted in a rise in the maximum power density from 9.5 to 12.37 mW cm⁻2. These results prove that the incorporation of nano-CaCO₃ as a filler in a Nafion matrix is practicable for DMFC applications.
... While eggshell calcium is well-known for its ability to prevent bone loss and osteoporosis by increasing bone density, calcium up-take and bone formation (Rovenský et al., 2003), ESM is more often used as supplement in osteoarthritis therapy. In case, a significant improvement of clinical symptoms has been reported in patients suffering from osteoarthritis (Danesch, 2014;Quintana et al., 2018). Interestingly, none of these clinical studies have associated the beneficial effect of ESM with an influence of ESM on bone formation. ...
Bone loss is a well-known phenomenon in the older population leading to increased bone fracture risk,morbidity, and mortality. Supplementation of eggshell membrane (ESM) is evaluated due to its possible appli-cation to prevent bone loss and usage in osteoporosis therapy. The similar organic chemical composition of ESMand human bone is described in detail as both mainly consist of collagen type I, chondroitin sulfate, dermatansulfate, hyaluronic acid and elastan. ESM and its components are reported to improve mineralization in bonetissue. In many studies ESM intake reduced pain in patients with joint disorders and reduced inflammatoryprocesses. Additionally, ESM improved calcium uptake in human cells. These findings in comparison with aclinical pilot study reporting pain reduction in osteoporotic patients and increased osteoblast activity in in vitroassays support ESM to be a beneficial supplement for bone health. In this systematic review we combinedchemical structure analysis with clinical studies to give a more comprehensive picture with novel explanations.
... The increased demand for eggs for consumption has large increases in the deposition of eggshells as environmental pollution worldwide and increase high management costs [7,8]. For example, the breaker plant in the USA used US$100,000 per year to dispose of eggshells into the environment (landfills) [9], Just Egg (Chilled Foods) Ltd, UK, used US$65,000 per year to dispose eggshells into the environment (landfills) [10], while the small-and medium-sized egg processing companies in Europe spent US$112,000 per year to dispose eggshells into the environment (landfills) [11]. ...
The increased demands for eggs for consumption have large increases in the deposition of eggshells as environmental pollution worldwide and increase high management costs. To reduce the management costs of eggshells, this work attempt was made to utilize eggshells as a source of CaCO3 for improving the hardness values and wear behavior of composite coating on mild steel. The composite coating was produced using 0, 5, 10, 15, and 20 g CaCO3-derived eggshell particles. The response surface method was used to determine the influences of applied load, sliding speed, and weight of CaCO3-derived eggshell particles on the wear behavior. The results have shown that Zn- CaCO3-derived eggshell particles composite coatings were successfully formed on mild steel. A 90.17% improvement of hardness values was obtained at Zn-20 g CaCO3-derived eggshell particles. The Zn-20 g CaCO3-derived eggshell particles sample has a lower friction coefficient. Wear rate increase with the increase in applied load and sliding speed. The optimum condition was obtained at CaCO3-derived eggshell particles (+ 1), applied load (− 0.35), and speed (− 0.58) with the lowest wear rate of 0.99 mm³/Nm. It has been recommended that the Zn-20 g CaCO3-derived eggshell particles composite coating of mild steel can be used in areas where wear and roller body are paramount.
... ESM suppresses the production of IL-1β and TNF-α inflammatory cytokines, subsiding inflammation (Hewlings et al., 2019). The double-blind and placebo study by Gil-Quintana et al. (2018) on the effect of daily intake of ESM supplement displayed shortterm effects within the first 3 days. Joint pain was significantly reduced, as was joint dysfunction. ...
Eggshell waste is among the most abundant waste materials coming from food processing technologies. Despite the unique properties that both its components (eggshell, ES, and eggshell membrane, ESM) possess, it is very often discarded without further use. This review article aims to summarize the recent reports utilizing eggshell waste for very diverse purposes, stressing the need to use a mechanochemical approach to broaden its applications. The most studied field with regards to the potential use of eggshell waste is catalysis. Upon proper treatment, it can be used for turning waste oils into biodiesel and moreover, the catalytic effect of eggshell-based material in organic synthesis is also very beneficial. In inorganic chemistry, the eggshell membrane is very often used as a templating agent for nanoparticles production. Such composites are suitable for application in photocatalysis. These bionanocomposites are also capable of heavy metal ions reduction and can be also used for the ozonation process. The eggshell and its membrane are applicable in electrochemistry as well. Due to the high protein content and the presence of functional groups on the surface, ESM can be easily converted to a high-performance electrode material. Finally, both ES and ESM are suitable for medical applications, as the former can be used as an inexpensive Ca²⁺ source for the development of medications, particles for drug delivery, organic matrix/mineral nanocomposites as potential tissue scaffolds, food supplements and the latter for the treatment of joint diseases, in reparative medicine and vascular graft producing. For the majority of the above-mentioned applications, the pretreatment of the eggshell waste is necessary. Among other options, the mechanochemical pretreatment has found an inevitable place. Since the publication of the last review paper devoted to the mechanochemical treatment of eggshell waste, a few new works have appeared, which are reviewed here to underline the sustainable character of the proposed methodology. The mechanochemical treatment of eggshell is capable of producing the nanoscale material which can be further used for bioceramics synthesis, dehalogenation processes, wastewater treatment, preparation of hydrophobic filters, lithium-ion batteries, dental materials, and in the building industry as cement.
... In recent years there has been an upswing with several small-scale start-up companies that have separated the membrane from waste eggshells at an industrial scale. For instance, Eggnovo (Spain) utilized calcium carbonate from eggshells for use in food, food supplements, cosmetics and pharmaceutical sectors [17]. American Dehydrated Foods (USA) in conjunction with Georgia Institute of Technology researchers has been developing a separation facility to recover calcium carbonate from eggshells for possible use in plastics in an effort to reduce petroleum-based products [15]. ...
Disposal of massive amounts of eggshells and seashells from processing industries is a challenge. In recent years, there has been a focus to reuse these waste resources in the production of new thermoplastic and thermoset polymer materials. This paper reviews eggshell and seashell production by country and provides a perspective on the quantity of bio-calcium carbonate that could be produced annually from these wastes. The achievements obtained from the addition of recycled bio-calcium carbonate fillers (uncoated/unmodified) in polymer composites with a focus on tensile strength, flexural strength and impact toughness are discussed. To improve compatibility between calcium carbonate (mineral and bio-based) fillers and polymers, studies on surface modifiers are reviewed. Knowledge gaps and future research and development thoughts are outlined. Developing novel and innovative composites for this waste material could bring additional revenue to egg and seafood processors and at the same time reduce any environmental impact.
Osteoarthritis (OA) is a chronic degenerative joint disease that causes chronic pain, swelling, stiffness, disability, and significantly reduces the quality of life. Typically, OA is treated using painkillers and non-steroidal anti-inflammatory drugs (NSAIDs). While current pharmacologic treatments are common, their potential side effects have prompted exploration into functional dietary supplements. Recently, eggshell membrane (ESM) has emerged as a potential functional ingredient for joint and connective tissue disorders due to its clinical efficacy in relieving joint pain and stiffness. Despite promising clinical evidence, the effects of ESM on OA progression and its mechanism of action remain poorly understood. This study evaluated the efficacy of Ovomet®, a powdered natural ESM, against joint pain and disease progression in a monosodium iodoacetate (MIA)-induced rodent model of OA in mice and rats. The results demonstrate that ESM significantly alleviates joint pain and attenuates articular cartilage destruction in both mice and rats that received oral supplementation for 5 days prior to OA induction and for 28 days thereafter. Interestingly, ESM significantly inhibited mRNA expression levels of pro-inflammatory cytokines including tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6), as well as inflammatory mediators, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase in the knee joint cartilage at the early stage of OA, within 7 days after OA induction. However, this effect was not observed in the late stage at 28 days after OA induction. ESM further attenuates the induction of protein expression for cartilage-degrading enzymes like matrix metalloproteinase (MMPs) 3 and 13, and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5), in the late-stage. In addition, MIA-induced reduction of the protein expression levels of cartilage components, cartilage oligomeric matrix protein (COMP), aggrecan (ACAN) and collagen type II α-1 chain (COL2α1), and cartilage extracellular matrix (ECM) synthesis promoting transcriptional factor SRY-Box 9 (SOX-9) were increased via ESM treatment in the cartilage tissue. Our findings suggest that Ovomet®, a natural ESM powder, is a promising dietary functional ingredient that can alleviate pain, inflammatory response, and cartilage degradation associated with the progression of OA.
Under the shell of a chicken egg are two opposed proteinaceous disulfide-rich membranes. They are fabricated in the avian oviduct using fibers formed from proteins that are extensively coupled by irreversible lysine-derived crosslinks. The intractability of these eggshell membranes (ESM) has slowed their characterization and their protein composition remains uncertain. In this work, reductive alkylation of ESM followed by proteolytic digestion led to the identification of a cysteine rich ESM protein (abbreviated CREMP) that was similar to spore coat protein SP75 from cellular slime molds. Analysis of the cysteine repeats in partial sequences of CREMP reveals runs of remarkably repetitive patterns. Module a contains a C-X4-C-X5-C-X8-C-X6 pattern (where X represents intervening non-cysteine residues). These inter-cysteine amino acid residues are also strikingly conserved. The evolutionarily-related module b has the same cysteine spacing as a, but has 11 amino acid residues at its C-terminus. Different stretches of CREMP sequences in chicken genomic DNA fragments show diverse repeat patterns: e.g. all a modules; an alternation of a-b modules; or an a-b-b arrangement. Comparable CREMP proteins are found in contigs of the zebra finch (Taeniopygia guttata) and in the oviparous green anole lizard (Anolis carolinensis). In all these cases the long runs of highly conserved modular repeats have evidently led to difficulties in the assembly of full length DNA sequences. Hence the number, and the amino acid lengths, of CREMP proteins are currently unknown. A 118 amino acid fragment (representing an a-b-a-b pattern) from a chicken oviduct EST library expressed in Escherichia coli is a well folded, highly anisotropic, protein with a large chemical shift dispersion in 2D solution NMR spectra. Structure is completely lost on reduction of the 8 disulfide bonds of this protein fragment. Finally, solid state NMR spectra suggest a surprising degree of order in intact ESM fibers.
An immunohistochemical analysis of the eggshell membranes shows the occurrence of type X collagen while type I collagen was not detected by using an appropriate monoclonal antibody with untreated shell membranes. A positive immuno-reaction for type I collagen was obtained after digestion of the shell membranes with pepsin. These observations indicate the possibility that type I collagen epitope was masked by type X collagen and that type X collagen may serve as an inhibitory boundary for biomineralization.
Sepsis is a systemic inflammatory response syndrome during infection. Therapeutic agents are essential to protect the host from sepsis. Ovocalyxin-36 (OCX-36) isa chicken eggshell membrane protein and shares protein sequence and gene organization homology with bactericidal permeability-increasing protein (BPI), lipopolysaccharide-binding protein (LBP) and palate, lung and nasal epithelium clone (PLUNC) proteins that play a major role in innate immune protection. We recently reported that OCX-36 binds to both lipopolysaccharide (LPS) and lipoteichoic acid (LTA) (Cordeiro et al., 2013, PLOS ONE 8, e84112), which is an important activity to neutralize endotoxins and non-endotoxin pyrogens during an inflammatory response. Here we investigated the immune modulating effects of OCX-36 and enzymatically digested OCX-36 (dOCX-36) in vitro and in a mouse model of endotoxemia. OCX-36 alone dose-dependently induced both TNF-α and nitric oxide (NO) production by RAW 264.7 macrophage cells, and this immunostimulatory effect was reduced by enzymatic digestion. In the presence of LPS, dOCX-36 was more effective than intact OCX-36 at reducing LPS-induced secretion of TNF-α from RAW 264.7 cells, but did not reduce NO production. In contrast, OCX-36 increased LPS-induced NO production, both in the presence and absence of FBS, PCR array analysis confirmed that OCX-36 and dOCX-36 differentially regulated genes involved in innate immunity, and dOCX-36 down-regulated the expression of genes involved in LPS signaling and inflammatory responses. In vivo, dOCX-36 was more effective at reducing LPS-induced inflammatory symptoms and inhibiting the local production of pro-inflammatory mediators in the small intestine. These resultssuggest that OCX-36 and OCX-36 derived peptides may differentially modulate innate immune responses, and support our hypothesisthat OCX-36 derived peptides have potential therapeutic applications in sepsis.
Collagen-like proteins have been found in the egg shell membranes of the hen. Materials similar to types I and V collagens were detected in each of the two layers of this membrane, the thick outer membrane and the thin inner membrane. Collagen was extracted by acid-pepsin digestion and isolated by differential salt precipitation. Identification of type-specific collagen-like material was established by coelectrophoresis on SDS-polyacrylamide gels using known collagen standards. These bands were susceptible to digestion by bacterial collagenase. From differential staining of the gels it was estimated that the ratio of collagen types I:V was approximately 100:1. Further confirmation of these biochemical results was obtained with immunofluorescence microscopy using type-specific antisera against chicken types I and V collagen with the indirect sandwich technique. Both the inner and outer shell membranes contained the two types of collagen. Within each membrane, the large, coarse 2.5-micron fibers contained predominantly type I collagen-like material, while type V collagen was mainly associated with the delicate narrower fibers of approximately 0.6-micron diameter. These tended to be concentrated in the inner membrane. At the electron microscopic level, both types of fibers were coated with glycoproteins that stained positively with ruthenium red. The deposition of these collagen-like substances by the hen oviduct on to the surface of the developing egg is an additional example of interstitial-type collagen synthesis and secretion by epithelial rather than by mesenchymal cells.
Investigation of eggshell membrane by amino acid analysis and radioimmunoassay established the presence of the crosslinking amino acids desmosine and isodesmosine at levels of 28 microgram/100 mg dry weight. Specific enzymatic and chemical hydrolysis indicates that these crosslinks are present as constituents of eggshell membrane protein and do not reflect the presence of small amounts of elastin. The eggshell membrane protein has approximately 10 times the level of lysine-derived aldehyde found in elastin.
The avian eggshell is a composite biomaterial composed of non-calcifying eggshell membranes and the overlying calcified shell matrix. The calcified shell forms in a uterine fluid where the concentration of different protein species varies between the initial, rapid calcification and terminal phases of eggshell deposition. The role of these avian eggshell matrix proteins during shell formation is poorly understood. The properties of the individual components must be determined in order to gain insight into their function during eggshell mineralization. In this study, we have identified lysozyme as a component of the uterine fluid by microsequencing, and used western blotting, immunofluorescence and colloidal-gold immunocytochemistry to document its localization in the eggshell membranes and the shell matrix. Furthermore, Northern blotting and RT-PCR indicates that there is a gradient to the expression of lysozyme message by different regions of the oviduct, with significant albeit low levels expressed in the isthmus and uterus. Lysozyme protein is abundant in the limiting membrane that circumscribes the egg white and forms the innermost layer of the shell membranes. It is also present in the shell membranes, and in the matrix of the calcified shell. Calcite crystals grown in the presence of purified hen lysozyme exhibited altered crystal morphology. Therefore, in addition to its well-known anti-microbial properties that could add to the protective function of the eggshell during embryonic development, shell matrix lysozyme may also be a structural protein which in soluble form influences calcium carbonate deposition during calcification.
The eggshell is an highly ordered structure deposited in the distal oviduct and composed of calcium carbonate and an organic matrix which is believed to influence its fabric. We have identified ovotransferrin as an 80 kDa matrix protein observed at high concentration in the uterine fluid at the initial stage of shell mineralization, by N-terminal sequencing and western blotting using monoclonal and polyclonal antibodies. It is present in extracts from demineralized eggshell and was localized by immunofluorescence in the eggshell membranes and mammillae, which are the sites of calcite nucleation. Northern blotting and RT-PCR demonstrated that ovotransferrin message was expressed in the proximal oviduct (magnum and white isthmus), and at a lower magnitude in the distal oviduct (red isthmus and uterus). Ovotransferrin was revealed by immunofluorescence in the tubular gland cells of the uterus. Calcium carbonate crystals grown in vitro in the presence of purified ovotransferrin showed large modifications of the calcite morphology. These observations and its presence in eggshell and membranes suggest a dual role for ovotransferrin, as a protein influencing nucleation and growth of calcite crystals and as a bacteriostatic filter to reinforce its inhibition of Salmonella growth in egg albumen.
Clinical measurement in both clinical research and clinical practice requires tools and techniques that are valid, reliable and responsive. Patient-centred self-reported measures provide opportunity to evaluate consequences of osteoarthritis, that are important and relevant to patients with the condition. The WOMAC and AUSCAN Indices are health status measurement questionnaires that are valid, reliable and responsive, easy to complete, simple to score and available in multiple language forms and scaling formats. They provide opportunities to capture patient relevant information, relating to the impact of interventions, in clinical research and clinical practice environments. WOMAC data have also contributed to the development of proposed definitions for responder criteria and state-attainment criteria in osteoarthritis.
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