Eggshell Membrane Research Articles

Covalent Organic Framework-Based Nanomotor for Multimodal Cancer Photo-Theranostics.

Developing efficient integrated diagnosis and treatment agents based on fuel-free self-movement nanomotors remains challenging in antitumor therapy. In this study, a covalent organic framework (COF)-based biomimetic nanomotor composed of polypyrrole (PPy) core, porphyrin-COF shell, and HCT116 cancer cell membrane coating is reported. Under near-infrared (NIR) light irradiation, the obtained mPPy@COF-Por can overcome Brownian motion and achieves directional motion through self-thermophoretic force generated from the PPy core. The HCT116 cancer cell membrane coating enables the nanomotor to selectively recognize the source cell lines and reduces the bio-adhesion of mPPy@COF-Por in a biological medium, endowing with this NIR light-powered nanomotor good mobility. More importantly, such multifunctional integration allows the COF-based nanomotor to be a powerful nanoagent for cancer treatment, and the high infrared thermal imaging/photoacoustic imaging/fluorescence trimodal imaging-guided combined photothermal/photodynamic therapeutic effect on HCT116 tumor cell is successfully achieved. The results offer considerable promise for the development of COF nanomotors with integrated imaging/therapy modalities in biomedical applications.

Comparative proteomics elucidates the potential mechanism of heritable carapace color of three strains Chinese mitten crab Eriocheir sinensis

The carapace coloration is important for the environmental adaptation and reproductive behaviors of crustaceans. We selected red, green and white three carapace color strains of Chinese mitten crab (Eriocheir sinensis) strains. These three carapace colors have stable heritability, but the mechanism for their coloration remains unclear.Through histological observations, we have found significant differences in the composition of pigment cells and pigments within the inner membrane of the three color strains, which may be one of the reasons for the color variation. The levels of various carotenoids in both the shell and inner membrane tissues of red and green strains were significantly higher than those of the white strain, while there was no significant difference between the red and green strains. Proteomics studies have identified 2, 034 and 947 different proteins in the shell and inner membrane, respectively. In the shell, there were 18, 13 and 43 differential proteins between red and white strains, green and white strains and green and red strains, respectively. In the inner membrane, there were 44, 24 and 16 differential proteins between red and white strains, green and white strains and green and red strains, respectively. It is clear that the deposited quantity of carotenoids affects the shell formation of three color strains. Some members of the hemocyanin family showed significant variation among different strains. The study yielded two crustacyanin proteins, which were extracted from both the shell and membrane. Of the two proteins, only Crustacyanin-A1 expression showed a difference between the red and green shells strains. In conclusion, these results indicated that the carapace color formation of E. sinensis may be accomplished through pigment binding proteins (PBPs) and pigment cells, which enhance the understanding of color formation mechanism for crustacean.

Fabrication of avian eggshell membrane derived dispersed collagen hydrogels for potential bone regeneration

Abstract Tissue engineering is emerging as an effective and alternate strategy for healing the impaired bones. This study reports the development of potential and novel bone tissue regenerating matrices from the avian eggshell membrane derived collagen dispersed in poly-(vinyl alcohol) (PVA). The concentration of the aqueous collagen dispersions within the PVA solution were varied from 0.5 % to 2 % (w/V); and the sols were subjected to varying freeze-thaw cycles to achieve gelation; leading to the fabrication of hydrogel matrices. The developed hydrogels were investigated for their physico-chemical characteristics through the ATR-FTIR, XRD, FESEM and biocompatibility measurements. The ATR-FTIR results showed the presence of amide A, amide I and hydroxyl groups; which were further reinstated by the XRD studies. The morphological and topological analysis of the different hydrogel groups was made through FESEM. It comprised of the measurements of pore dimensions ranging (3.98 ± 1.84 µm to 9.24 ± 5.55 µm), percentage porosity (47 %–97 %), and average surface roughness (21–39 µm); thus, indicating them to be analogous support systems for bone tissue regeneration. Further, L929 mouse fibroblasts grown over these support systems showed excellent cell viability, thus hinting towards its competitive features and application towards remedial bone regeneration.

Bio composite of porous hydroxyapatite and collagen extracted from eggshell membrane and Oreochromis niloticus fish skin for bone tissue applications

AbstractPorous hydroxyapatite (HA) was prepared by a chemical reaction between eggshell and phosphoric acid mixed with a foaming agent. Hydroxyapatite was composited with collagen prepared from a fermentation process of Acetobacter xylinum with eggshell membranes, extraction process of Oreochromis niloticus fish skin, and commercial‐grade collagen. Results of physical‐mechanical‐thermal properties of the bio composite materials yielded them as potential candidates for bone tissue engineering, drug delivery through the body, as well as tissue regeneration. The porous hydroxyapatite added foaming agent namely 64HA‐1NaHCO3‐500 and 64HA‐1NaHCO3‐600 having pore size and pore volume suitable for good adhering of collagen fiber types prepared from the fermentation process of Acetobacter xylinum of eggshell membrane and freeze‐drying process of Oreochromis niloticus. Because after reinforcing with collagen gave the highest compressive strength and three‐point bending strength values higher than HA samples without collagen three to five magnitudes. Furthermore, both bio composite samples served as positive thermal shock absorbers with significantly strong scanning electron microscope adhesion microstructures of collagen fibers within the porous HA matrices.

Evaluation of bone marrow-derived mesenchymal stem cells with eggshell membrane for full-thickness wound healing in a rabbit model.

Biomaterials capable of managing wounds should have essential features like providing a natural microenvironment for wound healing and as support material for stimulating tissue growth. Eggshell membrane (ESM) is a highly produced global waste due to increased egg consumption. The unique and fascinating properties of ESM allow their potential application in tissue regeneration. The wound healing capacity of bone marrow-derived mesenchymal stem cells (BM-MSCs), ESM, and their combination in rabbits with full-thickness skin defect (2 × 2cm2) was evaluated. Twenty-five clinically healthy New Zealand White rabbits were divided into five groups of five animals each, with group A receiving no treatment (control group), group B receiving only fibrin glue (FG), group C receiving FG and ESM as a dressing, group D receiving FG and BM-MSCs, and group E receiving a combination of FG, ESM, and BM-MSCs. Wound healing was assessed using clinical, macroscopical, photographic, histological, histochemical, hematological, and biochemical analysis. Macroscopic examination of wounds revealed that healing was exceptional in group E, followed by groups D and C, compared to the control group. Histopathological findings revealed improved quality and a faster rate of healing in group E compared to groups A and B. In addition, healing in group B treated with topical FG alone was nearly identical to that in control group A. However, groups C and D showed improved and faster recovery than control groups A and B. The macroscopic, photographic, histological, and histochemical evaluations revealed that the combined use of BM-MSCs, ESM, and FG had superior and faster healing than the other groups.

Permeability Model of Liquid Microcapsule Based on Multiple Linear Regression Method

The release rate of liquid core material from microcapsules is crucial for the surface properties of self-protective metal/liquid microcapsule composite plating coating. However, there is no method to accurately predict the release rate of microcapsules. In this paper, the permeability experiments of different shell membranes and core materials were carried out using the weight loss method, and the permeability model of liquid microcapsules was studied based on a multiple linear regression method. The results show that three-variable mathematical model C, including membrane porosity, the viscosity of core material, and membrane thickness is suitable to describe permeability, and the membrane thickness is the most significant influence factor. Additionally, the accuracy of model C was experimentally verified, and the error of the permeation rate is about 2.06% between predictive and experimental values.

Erythrocyte membrane-camouflaged and double-factor sequential delivery nanocarriers postpone the progression of osteoarthritis

Osteoarthritis is a commonly severe joint disease featured by the progressive loss of cartilage and reduction in lubricating synovial fluid, which currently lacks the effective treatment in clinic. Herein, we designed an erythrocyte membrane-camouflaged and double-factor sequential delivery of shell-core nanocarrier to deliver two diverse drugs. The interior core of nanocarriers was loaded with the transforming growth factor-β3 (TGF-β3), and subsequently modified with a natural anti-inflammatory drug of curcumin, and finally encapsulated with erythrocyte membrane as a protective and regulation shell. The as-prepared engineered nanocarriers were injected into joint capsule to play a dual function on inflammation modulation and cartilage remodeling. The curcumin absorbed on the internal surface of shell membrane was firstly released in large quantities to tune the inflammatory microenvironment whereas the TGF-β3 in the interior core was released subsequently with the nanoparticle degradation to alleviate chondrocyte senescence. Overall, our current work creates an engineered nanocarrier to control macrophage type and induce cartilage restoration on postponing the progression of osteoarthritis.

One‐Step Synthesis of Hydrogen‐Bonded Microcapsules for pH‐Triggered Protein Release

One‐step microfluidic approach in which interfacial complexation of hydrogen bonding polymers is utilized to prepare pH‐responsive microcapsules is presented. Using water‐in‐oil‐in‐oil‐in‐water (W1/O1/O2/W2) triple‐emulsion droplets as templates, it is shown that polymeric microcapsules with an aqueous core and a shell that consists of two complementary hydrogen bonding polymers, poly(propylene oxide)(PPO) and poly(methacrylic acid)(PMAA), can be prepared in a robust manner. The presence of a buffer layer enables controlled interfacial complexation of PMAA and PPO at the water/oil interface, followed by spontaneous dewetting of the oil droplet to result in hydrogen‐bonded microcapsules dispersed in aqueous media. In addition, it is demonstrated that the permeability of the PPO/PMAA membrane can be readily tuned by varying the molecular weight of PMAA. Furthermore, it is shown that the resulting PPO/PMAA microcapsules are stable at a high salt concentration (>1 m NaCl) unlike analogous capsules prepared through electrostatic complexation while they release the encapsulated protein above the critical pH of which the PMAA ionizes and results in disassembly of the shell membrane.

A drug-incorporated-microparticle-eggshell-membrane-scaffold (DIMES) dressing: A novel biomaterial for localised wound regeneration

Chronic wounds affect millions of people annually and have emotional and financial implications in addition to health issues. The current treatment for chronic wounds involves the repeated use of bandages and drugs such as antibiotics over an extended period. A cost-effective and convenient solution for wound healing is the development of drug-incorporated bandages. This study aimed to develop a biocompatible bandage made of drug-incorporated poly (lactic-co-glycolic acid) (PLGA) microparticles (MPs) and eggshell membrane (ESM) for cornea wound healing. ESM has desirable properties for wound healing and can be isolated from eggshells using acetic acid or ethylenediaminetetraacetic acid (EDTA) protocols. Fluorescein isothiocyanate-labelled Bovine Serum Albumin (FITC-BSA) was used as a model drug, and the PLGA MPs were fabricated using a solvent extraction method. The MPs were successfully attached to the fibrous layer of the ESM using NaOH. The surface features of the ESM samples containing MPs were studied using a field emission scanning electron microscope (FESEM) and compared with blank ESM images. The findings indicated that the MPs were attached to the ESM fibres and had similar shapes and sizes as the control MPs. The fibre diameters of the MPs samples were assessed using Fiji-ImageJ software, and no significant changes were observed compared to the blank ESM. The surface roughness, Ra values, of the MPs incorporated ESM samples were evaluated and compared to the blank ESM, and no significant changes were found. Fourier transform infrared (FTIR) spectroscopy was used to analyse the chemical Composition of the bandage, and the spectra showed that the FBM were effectively incorporated into the ESM. The FTIR spectra identified the major peaks of the natural ESM and the PLGA polymer in the bandage. The bandage was transparent but had a reduced visibility in the waterproof test card method. The bandage achieved sustained drug release up to 10 days and was found to be biocompatible and non-toxic in a chorioallantoic membrane (CAM) assay. Overall, the drug-incorporated PLGA MPs-ESM bandage has great potential for treating chronic wounds.

Preparation and characterization of propolis reinforced eggshell membrane/ GelMA composite hydrogel for biomedical applications

Gelatin methacrylate-based hydrogels (GelMA) were widely used in tissue engineering and regenerative medicine. However, to manipulate their various chemical and physical properties and create high-efficiency hydrogels, different materials have been used in their structure. Eggshell membrane (ESM) and propolis are two nature-derived materials that could be used to improve the various characteristics of hydrogels, especially structural and biological properties. Hence, the main purpose of this study is the development of a new type of GelMA hydrogel containing ESM and propolis, for use in regenerative medicine. In this regard, in this study, after synthesizing GelMA, the fragmented ESM fibers were added to it and the GM/EMF hydrogel was made using a photoinitiator and visible light irradiation. Finally, GM/EMF/P hydrogels were prepared by incubating GM/EMF hydrogels in the propolis solution for 24 h. After various structural, chemical, and biological characterizations, it was found that the hydrogels obtained in this study offer improved morphological, hydrophilic, thermal, mechanical, and biological properties. The developed GM/EMF/P hydrogel presented more porosity with smaller and interconnected pores compared to the other hydrogels. GM/EMF hydrogels due to possessing EMF showed compressive strength up to 25.95 ± 1.69 KPa, which is more than the compressive strength provided by GM hydrogels (24.550 ± 4.3 KPa). Also, GM/EMF/P hydrogel offered the best compressive strength (44.65 ± 3.48) due to the presence of both EMF and propolis. GM scaffold with a contact angle of about 65.41 ± 2.199 θ showed more hydrophobicity compared to GM/EMF (28.67 ± 1.58 θ), and GM/EMF/P (26.24 ± 0.73 θ) hydrogels. Also, the higher swelling percentage of GM/EMF/P hydrogels (343.197 ± 42.79) indicated the high capacity of this hydrogel to retain more water than other scaffolds. Regarding the biocompatibility of the fabricated structures, MTT assay results showed that GM/EMF/P hydrogel significantly (p-value < 0.05) supported cell viability. Based on the results, it seems that GM/EMF/P hydrogel could be a promising biomaterial candidate for use in various fields of regenerative medicine.

Stabilization of zirconia nanoparticles by collagen protein and calcium carbonate extracted from eggshell and its biodegradation, radical scavenging and mineralization activity

In this current study,collagen protein extracted from eggshell membrane and calcium carbonate (a main component of eggshell) are used as additive to enhance the stability and hardness of the zirconia crystals. Five different samples are prepared by adding membrane containing eggshell content as, 1 g, 2 g, 3 g, 4 g and 5 g in aqueous zirconium oxychloride sol. Phase purity is confirmed by XRD and FTIR. Phase pure and dense particles divulges the high hardness (∼1389 HV) and fracture toughness (12.89 MPa m1/2).FESEManalysis illustrates the formation of dense, well separated, non-agglomerated and spherical nanoparticles at 5 g of eggshell content. Eggshell works as a surfactant and stabilizer for formation of phase pure tetragonal spherical nanoparticles. Biodegradation study of optimized tetragonal zirconia (t-ZrO2)in phosphate buffered saline (PBS) presents that minor change in weight and hardness after 72 h of immersion. Antioxidant study shows the 96% of radical scavenging activity (RSA). In vitro bio-mineralization study shows the formation of new bone after 5 weeks. After 5 weeks all pores were filled and minerals were deposited on the surface of the scaffolds. SEM images confirms that eggshell-zirconia composite form new bone. So eggshell addition results in formation of phase pure t-ZrO2 nanoparticles with well-defined boundaries which exhibit higher hardness, fracture toughness, stability and enhanced antioxidant properties.

Biocompatibility and histological responses of eggshell membrane for dental implant-guided bone regeneration.

Guided bone regeneration (GBR) utilizing eggshell membrane (ESM) as a potential biomaterial for dental implant therapy augmentation was explored in this study. ESM, an environmentally friendly waste product, possesses collagen-rich characteristics. The biocompatibility and histological responses of ESM were investigated in a rat model. Twelve young adult Wistar rats were used in this study. ESM samples were implanted in subcutaneous and intramuscular pockets, and samples were collected at 48 hours, 4 weeks, and 8 weeks post-implantation. Histological analysis revealed the changes in ESM over time. Results showed that ESM maintained its structural integrity, induced a moderate cellular response, and exhibited slow degradation, indicating potential biocompatibility. However, the lack of organized collagen arrangement in ESM led to the formation of irregular and polymorphic spaces, allowing cell migration. Encapsulation of ESM by newly proliferating collagen fibers and multinucleated giant cells was observed at later time points, indicating a foreign body reaction. Crosslinking might improve its performance as a separation membrane, as it has the potential to resist enzymatic degradation and enhance biomechanical properties. In conclusion, ESM demonstrated biocompatibility, slow degradation, and lack of foreign body reaction. While not suitable as a complete separation membrane due to irregular collagen arrangement, further research involving crosslinking could enhance its properties, making it a viable option for guided bone regeneration applications in dental implant therapy. This study highlights the potential of repurposing waste materials for medical purposes and underscores the importance of controlled collagen structure in biomaterial development.

Congo red dye removal from aqueous solution by encapsulated eggshell membrane using central composite design

Eggshell is solid wastes produced from the food industry. Commonly, eggshell and its membrane used to treat water pollution such as heavy metal removal and also dye removal. Thus, on the treatment process, it’s was having the material handling problem, which was needed filtration method to remove out the adsorbent from the solution, and it was not efficient when commercializing in the wastewater treatment plant. Therefore, this study was using alcohol-sodium alginate (PVA/SA) to encapsulate the eggshell membrane, and this encapsulation method also would be able to enhance the performance of raw material on the dye adsorption. In this study, the eggshell membrane (ESM) are prepared in bead form, and the optimization of the bead are performed by the removal of congo red dye at several parameters such as pH, initial dye concentration, contact time, and dosage of ESM. The central composite design was selected to optimize the adsorption process by generating 30 runs of the experiment using software Design Expert® version 11. The Fourier Transforms Infrared Spectroscopy (FTIR) analysis and Brunauer-Emmett-Teller (BET) analysis are used to characterize the ESM bead. There was found out that main functional group on ESM bead contribute on adsorption process were carbonyl group, hydroxyl group and ESM bead also having the 42.74 nm of pore size. The optimize point for each parameter was 12.63mg/L of initial dye concentration, 6.12 g of ESM dosage, pH 2.19 and 12.83 min of contact time and the optimum congo red dye removal efficiency was 98.86 %.

Haemato-biochemical alterations associated with the use of eggshell membrane as a dressing material for full-thickness wounds in a rabbit model

Haemato-biochemical alterations associated with the use of eggshell membrane as a dressing material for full-thickness wounds in a rabbit model

Eggshell-Waste-Derived Calcium Acetate, Calcium Hydrogen Phosphate and Corresponding Eggshell Membranes

The development of innovative transformation techniques for various wastes generated by the agri-food industry is one of the goals of sustainable waste management oriented toward “zero-waste” approach. This includes the production of bulk and fine chemicals, bioactive compounds, enzymes and functional materials from various waste. The present research shows the possibility of the production of value-added products from eggshell waste using chemical transformation by acetic and o-phosphoric acid at a laboratory scale level. Eggshell-derived calcium acetate monohydrate and calcium hydrogen phosphate, both of high purity, as well as corresponding eggshell membranes as valuable by-products, were produced. Chemical transformation of 100 g of eggshell waste with 10% (w/v) acetic acid resulted in 111.41 ± 2.13 g of calcium acetate monohydrate and 2.90 ± 0.14 g of eggshell membranes, with a chemical yield in calcium acetate monohydrate of 79.16 ± 1.98%. On the other hand, the transformation of 100 g of eggshell waste in 15% (w/v) o-phosphoric acid yielded 77.06 ± 6.21 g of calcium hydrogen phosphate and 2.94 ± 0.08 g of eggshell membranes, with a chemical yield in calcium hydrogen phosphate of 71.36 ± 5.58%. Eggshell-derived calcium hydrogen phosphate met all prescribed criteria for food-grade additive, as determined by physicochemical analysis. On the other hand, calcium acetate monohydrate met the majority, except water insoluble and formic acid and oxidizable impurities content. Based on the obtained results, it can be concluded that the proposed eggshell waste transformation process by two different acids might be useful for the production of calcium acetate and calcium hydrogen phosphate as food-grade additives, as well as eggshell membranes as valuable by-products.

Eggshell Membrane Mediated Synthesis of CdO–NiO Nanocomposite for Multifunctional Applications

Eggshell Membrane Mediated Synthesis of CdO–NiO Nanocomposite for Multifunctional Applications

Synthesis and Assessment of Antimicrobial Composites of Ag Nanoparticles or AgNO3 and Egg Shell Membranes.

Engineering research has been expanded by the advent of material fusion, which has led to the development of composites that are more reliable and cost-effective. This investigation aims to utilise this concept to promote a circular economy by maximizing the adsorption of silver nanoparticles and silver nitrate onto recycled chicken eggshell membranes, resulting in optimized antimicrobial silver/eggshell membrane composites. The pH, time, concentration, and adsorption temperatures were optimized. It was confirmed that these composites were excellent candidates for use in antimicrobial applications. The silver nanoparticles were produced through chemical synthesis using sodium borohydride as a reducing agent and through adsorption/surface reduction of silver nitrate on eggshell membranes. The composites were thoroughly characterized by various techniques, including spectrophotometry, atomic absorption spectrometry, scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy, as well as agar well diffusion and MTT assay. The results indicate that silver/eggshell membrane composites with excellent antimicrobial properties were produced using both silver nanoparticles and silver nitrate at a pH of 6, 25 °C, and after 48 h of agitation. These materials exhibited remarkable antimicrobial activity against Pseudomonas aeruginosa and Bacillus subtilis, resulting in 27.77% and 15.34% cell death, respectively.

Methods of In Ovo and Ex Ovo Ostrich Embryo Culture with Observations on the Development and Maturation of the Chorioallantoic Membrane.

Culture of shell-free and windowed eggs for drug testing and other experiments has been perfected for smaller eggs such as those of chickens, where the developing blood vessels of the chorioallantoic membrane (CAM) become accessible for manipulative studies. However, due to the thickness and hardness of the ostrich egg shell, such techniques are not applicable. Using a tork craft mini rotary and a drill bit, we established windowed egg, in-shell-membrane windowed egg, and in-shell-membrane shell-free methods in the ostrich egg, depending on whether the shell membranes were retained or not. Concomitant study of the developing CAM revealed that at embryonic day 16 (E16), the three layers of the CAM were clearly delineated and at E25, the chorionic capillaries had fused with the epithelium while the CAM at E37 had reached maturity and the chorion and the allantois were both 3-4 times thicker and villous cavity (VC) and capillary-covering cells were well delineated. Both intussusceptive and sprouting angiogenesis were found to be the predominant modes of vascular growth in the ostrich CAM. Development and maturation of the ostrich CAM are similar to those of the well-studied chicken egg, albeit its incubation time being twice in duration.

Multicomponent SERS imprinted bio-membrane based on eggshell membrane for selective detection of spiramycin in water

A novel imprinting membrane (Ag/CuO/CNTs/Eggshell membrane -Molecularly Imprinted polymeric Membrances, ACCE-MIMs) based on surface enhanced Raman scattering (SERS) is successfully synthesized for the specific detection of spiramycin (SP). Firstly, CNTs and CuO are successively loaded on eggshell membrane (ESM) to increase the uniformity and sensitivity by virtue of the unique reticular structure of CNTs and excellent chemical enhancement of CuO. Through the abundant amino and carboxyl groups on ESM, Ag nanoparticles (Ag NPs) are anchored and growth on the surface of membrane by two in-situ reduction process and form a three-dimensional (3D) architecture. Eventually, the ACCE-MIMs are synthetized by silicon-based sol-gel method. With the help of imprinted polymers, the ACCE-MIMs present anti-interference performance during selective detection. The sensor presents a great liner relationship (R2 = 0.9965) between SP concentration and the Raman peak intensity when the concentration of SP ranges between 1 × 10−6 M ∼1 × 10−12 M. The limit of detection concentration is 1 × 10−12 M and the enhancement factor (EF) is 0.2956 × 107. The ACCE-MIMs exhibits sensitive detection property and this procedure provides a promising future for detection process.

Eggshell translucency in late-phase laying hens and its effect on egg quality and physiological indicators.

Eggshell translucency severely affects external egg quality, and variations in the eggshell or eggshell membrane are considered the structural basis of the trait. Research has shown that 1.85% additional mixed fatty acids in the diet would greatly decrease the occurrence of eggshell translucency. Only a few studies have examined the phenotypic regularity of eggshell translucency with the increasing age of hens. Therefore, two strains, 1139 Rhode Island Red-White (RIR-White) and 836 Dwarf Layer-White (DWL-White), were used, and from each strain, 30 hens each that consecutively laid translucent or opaque eggs at 67 wks of age were selected. Subsequently, eggshell translucency, internal quality and external quality of eggs, and total cholesterol, albumin, calcium binding protein and other physiological indicators related to lipid, lipoprotein, and calcium metabolisms at the 75th, 79th, and 83rd wks of age in the late phase of the laying cycle were determined. Results: (1) In terms of flocks, for both strains, the translucency scores of the translucent groups were significantly higher than those of the opaque groups (P < 0.05); in terms of individuals, 81.1% RIR-White and 82.8% DWL-White hens consecutively laid eggs of the same or similar translucency, indicating the stability of the trait with increasing hen age; (2) In RIR-White, the eggshell strength of the translucent group at 75 weeks was significantly higher than that of the opaque group (P < 0.05); in DWL-White, the eggshell membrane thickness of the translucent group at the 75th and 83rd weeks was significantly lower than that of the opaque group (P < 0.05); (3) Compared to the opaque groups, the translucent groups had lower total cholesterol content in both RIR-White and DWL-White, lower albumin content in DWL-White at the 79th weeks (P < 0.05), and higher calcium-binding protein (CALB1) in RIR-White at the 83rd weeks (P < 0.05). In summary, this study illustrates the stability of eggshell translucency in late-phase laying hens and provides a reference of physiological indicators for exploring the formation of translucent eggs.