Eggshell Membrane Research Articles

Eggshell Membrane Modulates the Gut Microbiota to Provide Health-Promoting Benefits

Eggshell Membrane Modulates the Gut Microbiota to Provide Health-Promoting Benefits

Further screening of SNP loci of eggshell translucency related genes and evaluation of genetic effects

Eggshell translucency is a widespread issue in the field of egg quality. Previous research has established that the heritability of eggshell translucency is relatively low or moderate. Scientists have also successfully identified SNP loci related to eggshell translucency on different chromosomes by using gene chips and single-variant GWAS. However, the specific impact of single or multiple genes on the trait of eggshell translucency remains unknown. In an effort to investigate this, we examined 170 SNPs associated with eggshell translucency obtained by our research group. We selected 966 half-sibling laying hens from 2 generations in 3 pure lines: Dwarf Layer-White, Rhode Island Red-White Strain, and Rhode Island Red. Eggs were collected from each hen over a period of 5 consecutive days, and eggshell translucency was measured using a grading method in which the hens were divided into 2 groups: an opaque group and a translucent group. We collected blood samples from the laying hens and extracted DNA. Time of flight mass spectrometry (TOF-MS) was used for genotyping to identify SNP loci that influence the trait of eggshell translucency. The results of our analysis revealed that using TOF-MS in 3 chicken strains, we were able to eliminate loci with low gene polymorphism, genetic effect contribution less than 1%, and deviation from Hardy-Weinberg equilibrium. Ultimately, 5 SNPs (Affx-50362599, rs15050262, rs312943734, rs316121113, and rs317389181) were identified on chromosomes 1, 5, and 19. Additionally, nine candidate genes (DCN, BTG1, ZFP92, POU2F1, NUCB2, FTL, GGNBP2, ACACA, and TADA2A) were found to be associated with these SNPs. No linkage disequilibrium relationship was observed between the 2 pairs of SNP loci on chromosomes 1 and 19. Based on previous studies on the formation mechanism of eggshell translucency, we hypothesize that NUCB2, FTL, and ACACA genes may be affecting the eggshell structure through different mechanisms, such as increase the water permeability or make thin of eggshell membrane, which promote moisture or part of other egg contents and ultimately lead to the formation of eggshell translucency. These findings validate and identify five SNP loci that regulate the translucency trait, and provide molecular markers for breeding non-translucent populations. Furthermore, this study serves as a reference for further investigation of the genetic regulatory mechanisms underlying eggshell translucency.

Time-Dependent Effect of Eggshell Membrane on Monosodium-Iodoacetate-Induced Osteoarthritis: Early-Stage Inflammation Control and Late-Stage Cartilage Protection.

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.

Natural Eggshell Membrane Attenuates Chondrocyte Inflammation and Surgically Induced Osteoarthritis in Rats

Natural Eggshell Membrane Attenuates Chondrocyte Inflammation and Surgically Induced Osteoarthritis in Rats

Stimulated Full-Thickness Cutaneous Wound Healing with Bioactive Dressings of Zinc and Cobalt Ion-Doped Bioactive Glass-Coated Eggshell Membranes in a Diabetic Rabbit Model.

Eggshell membrane-based biomedical applications have recently received great attention for their wound-healing properties. However, there are limited studies on diabetic wound healing. In this regard, we devised four types of composite eggshell membrane mats with nanoscale coatings of bioactive glass/Zn/Co-doped bioactive glass (ESM + BAG, ESM + ZnBAG, ESM + CoBAG, and ESM + ZnCoBAG) as wound-dressing materials for chronic nonhealing diabetic wounds. A detailed study of the physicochemical properties of the mats was conducted. In vitro studies demonstrated cytocompatibility and viability of human dermal fibroblasts on all four types of mats. The cells also attached finely on the mats with the help of cellular extensions, as evident from scanning electron microscopy (SEM) and rhodamine-phalloidin and Hoechst 33342 staining of cellular components. Endowed with bioactive properties, these mats influenced all aspects of full-thickness skin wound healing in diabetic animal model studies. All of the mats, especially the ESM + ZnCoBAG mat, showed the earliest wound closure, effective renewal, and restructuring of the extracellular matrix in terms of an accurate and timely accumulation of collagen, elastin, and reticulin fibers. Hydroxyproline and sulfated glycosaminoglycans were significantly (p < 0.01, p < 0.05) higher in ESM-ZnCoBAG-treated wounds in comparison to ESM-BAG-treated wounds, which suggests that these newly developed mats have potential as an affordable diabetic wound care solution in biomedical research.

Eggshell waste transformation to calcium chloride anhydride as food-grade additive and eggshell membranes as enzyme immobilization carrier

Abstract In continuation of our efforts to fully utilize eggshell waste (ESW), here we report the possibility of ESW transformation to calcium chloride (CaCl2) anhydride of food-grade additive purity and eggshell membranes (ESMs) as potential enzyme immobilization carriers. ESW chemical transformation by 5% (w/v) hydrochloric acid to CaCl2 solution and ESM completely devoid of the remnants of ESW calcium carbonate was performed in the constructed 15 L batch reactor during 4 h at room temperature, followed by separation of ESM from CaCl2 solution by filtration. ESW-derived CaCl2 solution containing the excess hydrochloric acid was neutralized by adding calcium hydroxide, concentrated to approximately 1/8th of volume, and spray dried. Separated ESM was washed with water and acetone, dried, and ground to a size of less than 0.5 mm. The ESW transformation process produced 102.42 ± 3.31 g of CaCl2 anhydrous and 2.48 ± 0.28 g of ESM per 100 g of ESW dry matter. ESW-derived CaCl2 fulfilled all criteria for food-grade additive, while obtained ESM showed their suitability for Burkholderia cepacia lipase immobilization by adsorption.

Concise Review on Integral Structure of Egg Shell Membrane

The article outlines various valuable applications for eggshell waste, including its use as a catalyst in biodiesel production to minimize pollutants, as an absorbent for heavy metals in wastewater, as a biomaterial for bone tissue replacement, and as a fertilizer and calcium supplement in various domains. It highlights the increasing research interest in exploring these applications for eggshell waste. This highlights the potential of the eggshell membrane (ESM) as a biomaterial for wound dressing due to its abundant availability and favourable properties. The study developed an extraction protocol for ESM and evaluated its physical, chemical, mechanical, and biological properties for wound dressing applications. Results showed that ESM retained its structure and composition after extraction, with promising characteristics such as optical transparency, porosity, fluid absorption, thermal stability, and mechanical strength. Biological studies confirmed its excellent biocompatibility with corneal cells, suggesting its potential for ophthalmic wound treatment and other biomedical applications, contributing to sustainable biomaterial development. The article discusses the formation and mineralization of calcareous eggs, primarily focusing on studies of chicken eggshells. It highlights areas of uncertainty such as the role of amorphous calcium carbonate and the molecules involved in eggshell formation. Additionally, it mentions the recent advancements in avian genomics and proteomics, which will aid in comparative studies of egg shell constituents across different bird species.

Biodegradable flexible triboelectric nanogenerator for winter sports monitoring

With the energy crisis and environmental pollution becoming a growing concern worldwide, the development of clean and renewable energy from the environment has become an imperative for human survival and development. However, the equipment used to harvest clean renewable energy is large, subject to environmental impacts and regional differences (such as wind, solar and tidal energy). In this study, a biodegradable eggshell membrane triboelectric nanogenerator (EM-TENG) is introduced for the purpose of harvesting low-frequency mechanical energy. A Wireless Intelligent Motion Monitoring System (WIMMS) has been created using EM-TENG. It includes a Bluetooth sensor terminal and an intelligent processing terminal for digital signal reception on a host computer. The EM-TENG can be attached to knee and ankle joints to monitor posture. Therefore, for real-time monitoring of joint and kinetic chain changes during land training of ice dance athletes, the intelligent ice dance land training aid system is important. As a wearable motion monitoring sensor, EM-TENGs application in intelligent motion monitoring, intelligent wearable devices and big data analytics is being promoted.

The effect of avian eggshell membrane structure on microbial penetration: A simulation study

Avian eggshells exhibit excellent antimicrobial properties. In this study, we conducted simulation experiments to explore the defense mechanisms of eggshell membranes with regards to their physical features. We developed a mathematical model for the movement of microorganisms and estimated their penetration ratio into eggshell membranes based on several factors, including membrane thickness, microbial size, directional drift, and attachment probability to membrane fibers. These results not only suggest that an eggshell membrane with multiple layers and low porosity indicates high antimicrobial performance, but also imply that the fibrous network structure of the membrane might contribute to effective defense. Our simulation results aligned with experimental findings, specifically in measuring the penetration time of Escherichia coli through the eggshell membrane. We briefly discuss the significance and limitations of this pilot study, as well as the potential for these results, to serve as a foundation for the development of antimicrobial materials.

White Isthmus Transcriptome Analysis Reveals the Mechanism of Translucent Eggshell Formation.

The presence of translucent eggshells is a type of egg quality issue that impacts egg sales. While many researchers have studied them, the exact mechanisms behind their formation remain unclear. In this study, we conducted a transcriptomic differential expression analysis of the isthmus region of the oviduct in both normal egg- and translucent egg-laying hens. The analysis revealed that differentially expressed gene pathways were predominantly concentrated in the synthesis, modification, and transport of eggshell membrane proteins, particularly collagen proteins, which provide structural support. These findings suggest that variations in the physical structure of the eggshell membrane, resulting from changes in its chemical composition, are the fundamental cause of translucent eggshell formation. This research provides a theoretical reference for reducing the occurrence of translucent eggs.

In situ green immobilization of palladium nanoparticles onto polydopamine-modified eggshell membrane for catalytic reduction of nitrophenols

4-Nitrophenol (4-NP) poses significant threats to both ecosystems and human health, making it essential and highly beneficial to take effective measures to mitigate its impact. Herein, eggshell membrane (ESM) with mussel-inspired polydopamine coatings (ESM-PDA) was constructed and utilized as a template for the immobilization of Pd NPs, resulting in ESM-PDA-Pd formation. The catalyst ESM-PDA-Pd exhibits exceptional performance in reducing 4-NP, and the TOF (turnover frequency) for ESM-PDA-Pd was determined to be 3.3 × 10−5 mmol·mg−1·min−1. Moreover, ESM-PDA-Pd demonstrates excellent catalytic activity in reducing other nitrophenols, including 2-nitrophenol (0.2196 min−1) and 2,4,6-nitrophenol (0.1177 min−1). Notably, the catalyst demonstrated brilliant recyclability, removing 92.4 % of 4-NP within 20 min after undergoing five consecutive runs. This underscores its potential for sustained and effective use in multiple cycles of nitrophenol reduction reactions. Furthermore, the measured Kapp in various water samples is as follows: ultrapure water (0.135 min−1) > tap water (0.109 min−1) > river water (0.083 min−1), indicating its significant potential for industrial applications.

Eggshell membrane powder reinforces adhesive polysaccharide hydrogels for wound repair

Multifunctional polysaccharide hydrogels with strong tissue adhesion, and antimicrobial and hemostatic properties are attractive wound healing materials. In this study, a chitosan-based hydrogel (HCS) was designed, and its properties were enhanced by incorporating oxidized eggshell membrane (OEM). Hydrogel characterization and testing results showed that the hydrogel had excellent antimicrobial properties, cytocompatibility, satisfactory adhesion properties on common substrates, and wet-state adhesion capacity. A rat liver injury model confirmed the significant hemostatic effect of the hydrogel. Finally, the ability of the hydrogel to promote wound healing was verified using rat skin wound repair experiments. Our findings indicate that HCS/OEM hydrogels with added eggshell membrane fibers have better self-healing properties, mechanical strength, adhesion, hemostatic properties, and biocompatibility than HCS hydrogels, in addition to having superior repair performance in wound repair experiments. Overall, the multifunctional polysaccharide hydrogels fabricated in this study are ideal for wound repair.

Eggshell membrane thickness and its contribution to total eggshell thickness for 13 waterbird species

Eggshell membrane thickness and its contribution to total eggshell thickness for 13 waterbird species

Valorization of Egg Shell Membrane as Protein Source in Soft Gel Capsules

Valorization of Egg Shell Membrane as Protein Source in Soft Gel Capsules

Exploration of novel DPP-IV inhibitory peptides from discarded eggshell membrane: An integrated in silico and in vitro study

Dipeptidyl peptidase-IV (DPP-IV) inhibitory peptides exhibit great potential to alleviate type II diabetes. This study aimed to prepare, identify and explore the molecular mechanism of DPP-IV inhibitory peptides from eggshell membrane (ESM), an abundant byproduct of the food industry, based on combining in silico and in vitro assessments. In silico evaluation of ten major ESM proteins revealed the great potential of ESM as a novel source of DPP-IV inhibitors. Moreover, the optimal combined-enzymatic strategy (alkaline protease + papain) for the efficient production of DPP-IV inhibitory peptides from ESM was designed by simulated proteolysis. Corresponding with in silico predictions, in vitro assessments confirmed that sodium sulfite assisted the screened dual enzymes led to better production of DPP-IV inhibitory peptides from ESM, with preferable total nitrogen recovery (86.61% ± 4.83%), DPP-IV inhibitory activity (64.61% ± 2.75%) and total antioxidant activity (215.27 ± 1.75 μg/mL). Due to the correlation between diabetes and oxidative stress, the antioxidant and DPP-IV inhibitory activities of the hydrolysate facilitate its amelioration of diabetes. Three novel DPP-IV inhibitory peptides with high abundance in ESM hydrolysates (YPEPPPQ, HDGADVS and VTDGQPH) were identified by LC-MS/MS, in silico characterization and molecular docking. These peptides were synthesized and confirmed to possess DPP-IV inhibitory activity, potentially relating to their stable binding with key residues within the S1, S2 pockets and catalytic triad of DPP-IV via hydrogen bonding and hydrophobic interaction. Overall, this research suggests an effective and time-saving strategy to investigate novel DPP-IV inhibitory peptides from ESM, as well as opens new opportunities for ESM application in antidiabetic foods.

Fabrication of Functional Gypsum Boards Using Waste Eggshells to Prevent Sick Building Syndrome

Eggshells can adsorb chemicals, but no studies regarding the adsorption of gaseous chemicals using eggshells or eggshell membranes have been reported. The purpose of this study was to apply chemical adsorption using eggshells to the maintenance of human health. Building materials containing eggshells may reduce the concentrations of toxic substances, such as formaldehyde, via the adsorption functions of eggshells. In the bending study, the strength of board-shaped gypsum-containing eggshells was not compromised when the content of eggshells within the gypsum was ≤10%. Compared to those of gypsum boards containing seashells, which comprise calcium carbonate, similar to eggshells, gypsum containing eggshells displayed a higher strength. In the adsorption study, board-shaped gypsum containing eggshells placed inside a sealed box rapidly decreased the formaldehyde concentration. A gypsum board with an eggshell content of ≥5% could limit the formaldehyde concentration to ≤0.08 ppm. Furthermore, the results were compared with those of adsorption studies using plasterboard mixed with other natural materials. Eggshells displayed excellent functionalities as novel formaldehyde adsorbents.

Harnessing potential of avian eggshell membrane derived collagen hydrolysate for bone tissue regeneration.

Natural bone grafts are the highly preferred materials for restoring the lost bone, while being constrained of donor availability and risk of disease transmission. As a result, tissue engineering is emerging as an efficacious and competitive technique for bone repair. Bone tissue engineering (TE) scaffolds to support bone regeneration and devoid of aforesaid limitations are being vastly explored and among these the avian eggshell membrane has drawn attention for TE owing to its low immunogenicity, similarity with the extracellular matrix, and easy availability. In this study, the development of bone ingrowth support system from avian eggshell membrane derived collagen hydrolysates (Col-h) is reported. The hydrolysate, cross-linked with glutaraldehyde, was developed into hydrogels with poly-(vinyl alcohol) (PVA) by freeze-thawing and further characterized with ATR-FTIR, XRD, FESEM. The biodegradability, swelling, mechanical, anti-microbial, and biocompatibility evaluation were performed further for the suitability in bone regeneration. The presence of amide I, amide III, and -OH functional groups at 1639cm- 1,1264cm- 1, and 3308cm- 1 respectively and broad peak between 16°-21° (2θ) in XRD data reinstated the composition and form. The maximum ratio of Col-h/PVA that produced well defined hydrogels was 50:50. Though all the hydrogel matrices alluded towards their competitive attributes and applicability towards restorative bone repair, the hydrogel with 40:60 ratios showed better mechanical strength and cell proliferation than its counterparts. The prominent E. coli growth inhibition by the hydrogel matrices was also observed, along with excellent biocompatibility with MG-63 osteoblasts. The findings indicate strongly the promising application of avian eggshell-derived Col-h in supporting bone regeneration.

The Chicken Egg: An Advanced Material for Tissue Engineering.

The chicken egg, an excellent natural source of proteins, has been an overlooked native biomaterial with remarkable physicochemical, structural, and biological properties. Recently, with significant advances in biomedical engineering, particularly in the development of 3D in vitro platforms, chicken egg materials have increasingly been investigated as biomaterials due to their distinct advantages such as their low cost, availability, easy handling, gelling ability, bioactivity, and provision of a developmentally stimulating environment for cells. In addition, the chicken egg and its by-products can improve tissue engraftment and stimulate angiogenesis, making it particularly attractive for wound healing and tissue engineering applications. Evidence suggests that the egg white (EW), egg yolk (EY), and eggshell membrane (ESM) are great biomaterial candidates for tissue engineering, as their protein composition resembles mammalian extracellular matrix proteins, ideal for cellular attachment, cellular differentiation, proliferation, and survivability. Moreover, eggshell (ES) is considered an excellent calcium resource for generating hydroxyapatite (HA), making it a promising biomaterial for bone regeneration. This review will provide researchers with a concise yet comprehensive understanding of the chicken egg structure, composition, and associated bioactive molecules in each component and introduce up-to-date tissue engineering applications of chicken eggs as biomaterials.

Anti-inflammatory effects of eggshell membrane hydrolysates on carrageenan-induced rat

Background: Eggshells are one of the most common culinary wastes, yet using eggshell membrane detritus in medical treatments remains largely unexplored. Objective: This study aims to assess the anti-inflammatory potential of eggshell membrane hydrolysate. Method: Eggshell membrane hydrolysate was prepared using acid-base hydrolysis. The intraplantar carrageenan-induced inflammation model was used to test the anti-inflammatory effect. The experimental group consisted of a control, comparison (4.5 mg/kg BW diclofenac-Na, 4.5 mg/kg BW hyaluronic acid, and 45 mg/kg BW collagen), and eggshell membrane hydrolysates (9, 22.5, 45 and 90 mg/kg BW). Observations of inflammation, percentage of inflammation, and percentage of inhibition of inflammation were measured every 30 minutes for 180 minutes. Statistical analysis was performed using the t-test and also trendline analysis. Result: The findings revealed that collagen and hyaluronic acid demonstrated the ability to suppress inflammation compared to controls (p &lt; 0.05). Compared to the control group, the group that received eggshell membrane hydrolysate at doses of 9, 22.5, 45, and 90 mg/kg BW also exhibited inhibition of inflammation (p &lt; 0.05). Furthermore, increasing the dosage of eggshell membrane hydrolysate correlated with an increase in the percentage of inflammation inhibition in carrageenan-induced rats. Conclusion: Eggshell membrane hydrolysate has the potential to act as an anti-inflammatory agent.

Eggshell membrane as a novel and green platform for the preparation of highly efficient and reversible curcumin-based colorimetric sensor for the monitoring of chicken freshness

Herein, for the very first time, we report a paper-like biomass, eggshell membrane (ESM), as a suitable platform for the fabrication of a colorimetric sensor (E-Cot). Green ethanolic extract, curcumin (CUR), was used as a sensing material to coat with the ESM. The present E-Cot effectively changed its color (yellow to red) in the real-time monitoring for chicken spoilage. The E-Cot exhibits barrier properties due to its inherent semi-permeability characteristics. Interestingly, the E-Cot showed a significant change in total color difference value (ΔE, 0 days - 0.0-39.6, after 1 day – 39.6-42.1, after 2 days – 42.1-53.6, after 3 days- 53.6-60.1, and after 4 days – 60.1-66.3, detectable by the naked eye) in the real-time monitoring for chicken freshness. In addition, the present E-Cot smart colorimetric sensor is reversible with a change in pH, and the sensor can be reused. Further, the hydrophobic nature of the E-Cot was confirmed by water contact angle analysis (WCA, contact angle of 101.21 ± 8.39). Good antibacterial, barrier, and optical properties of the present E-Cot were also found. Owing to the advantages such as green, efficient, cost-effective, biodegradable, reusable, sustainable, and simple preparation, we believe that the present E-Cot would be a more attractive candidate.