Eggshell membrane as a regenerative material in alveolar bone grafting in combination with advanced platelet rich fibrin.

Abstract: The eggshell and the eggshell membrane (ESM) are significant by-products of the poultry industry and are being utilized for various valuable purposes in health care, like soft tissue healing and pain alleviation. The aim and objective of our study are to assess the effect of the eggshell membrane on alveolar bone regeneration after tooth extraction. A total of 40 extraction sockets (bilateral) among 20 patients were assessed clinically for healing, and radiographic parameters of bone density and socket volume were assessed on CBCT at baseline, 3 months, and 6 months. Advanced platelet-rich fibrin was created from 5 ml of autologous blood from the patient and centrifuged for 15 minutes at 1500 RPM/168 RCF. The commercially available powdered form of egg shell membrane was used in the study. Based on the randomized allotment (coin-flip), A-PRF alone or A-PRF mixed with eggshell membrane was placed inside the extraction socket and was stabilized using 3-0 silk sutures. It was ob-served that wound healing was uneventful in all 20 patients. No evidence of dry sockets or allergic reactions was noted in any patient. Statistical analysis was done using the un-paired t-test and Mann-Whitney U test with SPSS version 20.0. P<0.05 was considered significant. On comparison of the mean bone density at baseline, 3 months, and 6 months, the socket density in the eggshell with the PRF group was higher compared to the control group. To conclude, eggshell membrane has good regenerative properties and excellent osteogenic capacity; therefore, it could be a useful graft due to its low cost, abundant availability, and simple application.

On the Structure and Role of Avian Eggshells: A 31P, 1H, and 13C Solid-State NMR Study.

The eggshell is a composite and highly ordered structure formed by biomineralization. Besides other functions, it has a vital and intricate role in the protection of an embryo from various potentially harsh environmental conditions. Solid-state nuclear magnetic resonance (SSNMR) has been used for detailed structural investigations of the chicken, tinamou, and flamingo eggshell materials. 31P NMR spectra reveal that hydroxyapatite and ß-tricalcium phosphate in the ratio 3:2 represent major constituents of phosphate species in the eggshells. All three eggshells exhibit similar spectra, except for the line widths, which implies different structural order of phosphate species in the chicken, tinamou, and flamingo eggshells. 1H NMR spectra for these materials are comparable, differentiating overlapped peaks in three spectral regions at around 7, 4-5, and 1-2 ppm. These spectral regions have been attributed to protons from NH or CaHCO3, water, and possibly isolated monomeric water molecules or hydroxyl groups in calcium-deficient hydroxyapatite. 1H-13C CP MAS NMR revealed the presence of organic matter in the form of lipids and proteins. Two overlapped resonances in the carbonyl region at around 173 and 169 ppm are assigned to the carbonyls of the peptide bonds and the bicarbonate unit in calcite, respectively. Fourier-transform infrared spectroscopy (FTIR) spectra confirmed the presence of structural units detected in the NMR spectra.

An innovative protocol to increase egg production of chicken layers.

This study investigated the effects of different doses of limestone, light durations, light intensities, and vitamins on both the productive performance and egg quality. The study utilized two rearing houses (control and treatment), each accommodating 75000 Lohmann Brown Classic chicks reared in open-sided rearing cages from one day old until they reached 89 weeks of age. Throughout the laying period, the hens were subjected to a specific light regimen (light = 14 h; dark = 10 h a day). At the end of experiment, the treatment group displayed significant (p<0.05) differences compared to the control group across various parameters. Notably, the treatment group exhibited lower daily feed intake (treatment: 112 g/bird vs control: 115 g/bird), 9.6% higher egg production (treatment: 78.5% vs control: 68.9%), lower body weight (treatment: 2057 g vs control: 2073 g), lower feed conversion ratio (FCR)/egg (treatment: 1.44 vs control: 1.69), higher egg weight (treatment: 69.4 g vs control: 68.5 g), greater egg mass (treatment: 56.14 vs control: 48.76), greater shell thickness (treatment: 3.52 mm vs control: 3.44 mm), and greater shell weight (treatment: 9.3 g vs control: 8.79 g). However, the albumin weight, yolk weight, yolk diameter, shape index, and Haugh units (HU) were not significantly (p˃0.05) affected after 75 weeks of treatment when compared with those of the control group. Therefore, this study is the first of its kind to demonstrate that different ratios of limestone, different durations and intensities of light, and different vitamin supplementation doses in the treatment group (subjected to the novel rearing recommendations described in this study) may yield a profit of 180,541 USD, exceeding the baseline profit of the control group (subjected to conventional rearing methods).

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-1ß (IL-1ß), 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.

Identification and Characterization of the miRNA Transcriptome Controlling Green Pigmentation of Chicken Eggshells.

Green eggs are mainly caused by inserting an avian endogenous retrovirus (EVA-HP) fragment into the SLCO1B3 gene. Although the genotypes for this insertion allele are consistent, eggshell color (ESC) may vary after a peak laying period; light-colored eggs are undesired by consumers and farmers and result in financial loss, so it is necessary to resolve this problem. miRNAs are small non-coding RNAs that exert essential functions in animal development and diseases. However, the regulatory miRNAs and detailed molecular mechanisms regulating eggshell greenness remain unclear. In the present study, we determined the genotype of green-eggshell hens through the detection of a homozygous allele insertion in the SLCO1B3 gene. The shell gland epithelium was obtained from green-eggshell hens that produced white and green shell eggs to perform transcriptome sequencing and investigate the important regulatory mechanisms that influence the ESC. Approximately 921 miRNAs were expressed in these two groups, which included 587 known miRNAs and 334 novel miRNAs, among which 44 were differentially expressed. There were 22 miRNAs that were significantly upregulated in the green and white groups, respectively, which targeted hundreds of genes, including KIT, HMOX2, and several solute carrier family genes. A Gene Ontology enrichment analysis of the target genes showed that the differentially expressed miRNA-targeted genes mainly belonged to the functional categories of homophilic cell adhesion, gland development, the Wnt signaling pathway, and epithelial tube morphogenesis. A KEGG enrichment analysis showed that the Hedgehog signaling pathway was significantly transformed in this study. The current study provides an overview of the miRNA expression profiles and the interaction between the miRNAs and their target genes. It provides valuable insights into the molecular mechanisms underlying green eggshell pigmentation, screening more effective hens to produce stable green eggs and obtaining higher economic benefits.

A Network of Circular RNA and MicroRNA Sequencing Provides Insights into Pigment Deposition of Changshun Blue Eggshell Chickens.

Eggshell color plays important biological roles and attracts the attention of both egg retailers and researchers. However, whether non-coding RNAs are involved in pigment deposition among different eggshell colors remains unknown. In this study, RNA sequencing was used to analyse the uterine gland transcriptome (CircRNA and miRNA) of Changshun chicken blue-shell hens producing four different eggshell color eggs including dark blue PK(DB) and light blue (LB), dark brown and greenish (between blue and pink, DP) and pink (p). We found that miR-192-x, targeting SLC16a7, was expressed in DB, DP, and LB groups compared with the PK group, which indicates that miR-192-x may play a role in the blue eggshell color. KEGG and GO analyses showed that the "metabolic pathways" with targeted genes such BLVRA and HMOX1 were detected in dark and light blue color eggshell chickens, which confirms the different ratios of biliverdin and HO-1 involved in the deposition of blue color. As annotated by connectivity analysis, RASGRF1 and RASGRF2, belonging to the RASGRF family, are involved in the Ras signaling pathway, which plays an important role in cell growth, differentiation, metastasis and apoptosis. Our findings enrich the database of circRNA, miRNAs and genes for chicken uterine tissue, which will be useful in accelerating molecular selection for blue eggshell color layers.

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.

3D Printed Eggshell Microparticle-Laden Thermoplastic Scaffolds for Bone Tissue Engineering.

Three-dimensional (3D) printing, an additive manufacturing technique, is increasingly used in the field of tissue engineering. The ability to create complex structures with high precision makes the 3D printing of this material a preferred method for constructing personalized and functional materials. However, the challenge lies in developing affordable and accessible materials with the desired physiochemical and biological properties. In this study, we used eggshell microparticles (ESPs), an example of bioceramic and unconventional biomaterials, to reinforce thermoplastic poly(ε-caprolactone) (PCL) scaffolds via extrusion-based 3D printing. The goal was to conceive a sustainable, affordable, and unique personalized medicine approach. The scaffolds were fabricated with varying concentrations of eggshells, ranging from 0 to 50% (w/w) in the PCL scaffolds. To assess the physicochemical properties, we employed scanning electron microscopy, Fourier-transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, and X-ray diffraction analysis. Mechanical properties were evaluated through compression testing, and degradation kinetics were studied through accelerated degradation with the remaining mass ranging between 89.4 and 28.3%. In vitro, we evaluated the characteristics of the scaffolds using the MC3T3-E1 preosteoblasts over a 14 day period. In vitro characterization involved the use of the Alamar blue assay, confocal imaging, and real-time quantitative polymerase chain reaction. The results of this study demonstrate the potential of 3D printed biocomposite scaffolds, consisting of thermoplastic PCL reinforced with ESPs, as a promising alternative for bone-graft applications.

Multilevel sensor for monitoring external and internal environment of eggs.

Although it is well known that incubation environment has a great influence on embryogenesis and post-hatching performance of birds, not much is known about how external thermal, sound and light stimuli are isolated by eggshells and perceived by embryos. In this context, this study aimed to develop, calibrate and evaluate a multilevel sensor for integrated monitoring of the external (incubator) and internal environment of eggs. The variables of interest for the external environment were air temperature and relative humidity. For the internal environment, shell temperature, internal temperature, luminosity and sound pressure level were considered. The sensor was developed with an ATmega328 microcontroller, in open-source prototyping, using electronic components which are compatible with the egg's physical structure. Calibrations were carried out in a controlled environment, comparing the multilevel sensor with commercial equipment, obtaining coefficients of determination of R 2 > 0.90 for all variables studied. The multilevel sensor was also validated, simulating a commercial incubation situation and comparing eggs with 2 shell colors (white and brown) and internal volume (intact and empty). Validation results showed that white-shelled eggs insulate less external light (P < 0.001) and full eggs presented higher internal temperatures, greater light and lower sound pressure levels compared to empty eggs (P < 0.001). The multilevel sensor developed here is an innovative proposal for monitoring, simultaneously and in real time, different variables of interest in the commercial incubation environment.

Genome-wide association analysis of eggshell color of an F2 generation population reveals candidate genes in chickens.

Eggshell color is an important visual characteristic that affects consumer preferences for eggs. Eggshell color, which has moderate to high heritability, can be effectively enhanced through molecular marker selection. Various studies have been conducted on eggshell color at specific time points. However, few longitudinal data are available on eggshell color. Therefore, the objective of this study was to investigate eggshell color using the Commission International de L'Eclairage L*a*b* system with multiple measurements at different ages (age at the first egg and at 32, 36, 40, 44, 48, 52, 56, 60, 66, and 72 weeks) within the same individuals from an F2 resource population produced by crossing White Leghorn and Dongxiang Blue chicken. Using an Affymetrix 600 single nucleotide polymorphism (SNP) array, we estimated the genetic parameters of the eggshell color trait, performed genome-wide association studies (GWASs), and screened for the potential candidate genes. The results showed that pink-shelled eggs displayed a significant negative correlation between L* values and both a* and b* values. Genetic heritability based on SNPs showed that the heritability of L*, a*, and b* values ranged from 0.32 to 0.82 for pink-shelled eggs, indicating a moderate to high level of genetic control. The genetic correlations at each time point were mostly above 0.5. The major-effect regions affecting the pink eggshell color were identified in the 10.3-13.0 Mb interval on Gallus gallus chromosome 20, and candidate genes were selected, including SLC35C2, PCIF1, and SLC12A5. Minor effect polygenic regions were identified on chromosomes 1, 6, 9, 12, and 15, revealing 11 candidate genes, including MTMR3 and SLC35E4. Members of the solute carrier family play an important role in influencing eggshell color. Overall, our findings provide valuable insights into the phenotypic and genetic aspects underlying the variation in eggshell color. Using GWAS analysis, we identified multiple quantitative trait loci (QTLs) for pink eggshell color, including a major QTL on chromosome 20. Genetic variants associated with eggshell color may be used in genomic breeding programs.

Physico-chemical and biological characterization of synthetic and eggshell derived nanohydroxyapatite/carboxymethyl chitosan composites for pulp-dentin tissue engineering.

Hybrid nanohydroxyapatite/carboxymethyl chitosan (nHAp-CMC) scaffolds have garnered significant attention in the field of regenerative engineering. The current study comparatively analyzed the physicochemical and biological properties of synthetic nanohydroxyapatite (SnHA)- and eggshell-sourced nanohydroxyapatite (EnHA)- based CMC biocomposites for pulp-dentin regeneration. EnHA and CMC were synthesized through a chemical process, whereas SnHA was commercially obtained. Composite scaffolds of SnHA-CMC and EnHA-CMC (1:5 w/w) were prepared using freeze-drying method. All biomaterials were characterized by FTIR, micro-Raman, XRD, HRSEM-EDX, and TEM analyses, and their in vitro bioactivity was assessed by immersing them in simulated body fluid for 21 days. The biological properties of the composite scaffolds were evaluated by assessing cytocompatibility using MTT assay and biomineralization potential by analyzing the odontogenic gene expressions (ALP, DSPP, DMP-1 and VEGF) in human dental pulp stem cells (DPSCs) using RT-qPCR method. Characterization studies revealed that EnHA displayed higher crystallinity and superior surface morphology compared to SnHA. The composite scaffolds showed a highly interconnected porous microstructure with pore sizes ranging between 60 and 220 µm, ideal for cell seeding. All tested materials, SnHA, EnHA, and their respective composites, displayed high cytocompatibility, increased ALP activity and degree of mineralization with significant upregulation of odontogenic-related genes on DPSCs (p < 0.05). Nevertheless, the odontogenic differentiation potential of EnHA-CMC on DPSCs was significantly higher when compared to SnHA-CMC. The findings from this study highlight the potential of EnHA-CMC as a promising candidate for pulp-dentin engineering.

Biomimetic dentin remineralization using eggshell derived nanohydroxyapatite with and without carboxymethyl chitosan - An in vitro study.

The objective of this study was to evaluate the synergistic effect of eggshell-derived nanohydroxyapatite (EnHA) and carboxymethyl chitosan (CMC) in remineralizing artificially induced dentinal lesions. EnHA and CMC were synthesized using simple chemical processes and characterized using FTIR, XRD, HRSEM-EDX, TEM, DLS and TGA/DTA analyses. A total of 64 pre-demineralized coronal dentin specimens were randomly subjected to following treatments (n = 16):artificial saliva (AS), EnHA, CMC, and EnHA-CMC, followed by pH cycling for 7 days. HRSEM-EDX, Vickers-indenter, and micro-Raman analyses were used to assess surface-topography, microhardness, and chemical analysis, respectively. All tested materials demonstrated non-cytotoxicity when assessed on hDPSCs using MTT assay. FTIR, XRD and thermal analyses confirmed the characteristics of both EnHA and CMC. EnHA showed irregular rod-shaped nanoparticles (30-70 nm) with the presence of Ca,P,Na, and Mg ions. Dentin treated with EnHA-CMC exhibited complete tubular occlusion and highest microhardness whereas the AS group revealed the least mineral deposits (p < 0.05). No significant differences were observed between EnHA and CMC groups (p > 0.05). In addition, molecular conformation analysis revealed peak intensities in collagen's polypeptide chains in dentin treated with CMC and EnHA-CMC, whereas other groups showed poor collagen stability. The results highlighted that EnHA-CMC aided in rapid and effective biomineralization, suggesting its potential as a therapeutic solution for treating dentin caries.

High albumen height by expression of GALNT9 and thin eggshell by decreased Ca2+ transportation caused high hatchability in Huainan partridge chicken.

Hatchability could be quite different among individuals of indigenous chicken breed which might be affected by the egg quality. In this study, hatchability was individually recorded among 800 forty-wk-old Huainan partridge chickens. The chickens were then divided into high and low hatchability groups (HH and LH group) with 50 birds in each group. Egg quality was further determined in the 2 groups. Eight birds from each group were selected for slaughtering and tissue, responsible for egg formation, collection for structure observation by staining and candidate gene expression by transcriptome analysis. The hatchability in HH was 100% and 61.18% in LH. The eggshell thickness and shell strength were significantly lower, while the albumen height and Haugh unit were significantly higher in HH group than those in LH group (P < 0.05). The magnum weight and index, and the expression of polypeptide N-acetylgalactosaminyltransferase 9 (GALNT9), which responsible for thick albumen synthesis, in HH group were also significantly higher than that of LH group (P < 0.05). Compared with the LH group, there were 702 differentially expressed genes (DEGs) in HH group, of which 402 were up-regulated and 300 were down-regulated. Candidate genes of calbindin 1 (CALB1) and solute carrier family 26 member 9 (SLC26A9), which regulate calcium signaling pathway so as to affect Ca2+ transportation, exhibited significant high and low expression, respectively, in HH group compared to those in LH group (P < 0.05). Therefore, indigenous chicken with high expression of GALNT9 in magnum to form thick albumen to provide more protein for embryo, while high CALB1 and low expression of SLC26A9 to decrease Ca2+ transportation so as to form a thinner eggshell and provide better gas exchange during embryo development.

Does Nesting Material Affect Wood Duck (Aix sponsa) Nest Box Selection, Reproduction, and Eggshell Bacteria?

Wood Ducks (Aix sponsa) are secondary cavity nesters that use natural cavities and artificial nest boxes, the latter of which has been attributed to the recovery of populations across the southeastern US. Continual use of these boxes results in a buildup of bacteria, parasites, and other pathogens. To avoid the accumulation of these deleterious organisms, best management practices include the occasional removal of old nesting material (i.e., wood shavings) and replacement with fresh wood shavings. No studies have been performed on the effects of shaving material on nest box selection, nest success, and bacterial growth. We monitored 142 and 111 nest boxes in Florida and Georgia, USA, respectively, and filled a random sample with aspen or cedar shavings. We then swabbed the surface of 144 and 150 eggs during 2020 and 2021, respectively, to screen for culturable bacteria. We detected no effect of shaving type on nest box selection, nest success, or egg surface bacterial growth. We found 3-8 bacterial colony types (1-123 colony-forming units [CFU]/box) and 1-8 bacterial colony types (3-382 CFU/box) among the Georgia and Florida samples, respectively. We detected no effect from shaving type on Wood Duck reproduction or bacterial growth in the sampled nest boxes. We concluded that both shaving types are suitable nesting materials for box-nesting Wood Duck populations and the continued use of either would be a reasonable decision for managers.

Synergistic effect of phytic acid and eggshell bio-fillers on the dual-phase fire-retardancy of intumescent coatings applied on cellulosic substrates.

Cellulosic substrates, including wood and thatch, have become icons for sustainable architecture and construction, however, they suffer from high flammability because of their inherent cellulosic composition. Current control measures for such hazards include applying intumescent fire-retardant (IFR) coatings that swell and form a char layer upon ignition, protecting the underlying substrate from burning. Typically, conventional IFR coatings are opaque and are made of halogenated compounds that release toxic fumes when ignited, compromising the roofing's aesthetic value and sustainability. In this work, phytic acid, a naturally occurring phosphorus source extracted from rice bran, was used to synthesize phytic acid-based fire-retardants (PFR) via esterification under reflux, along with powdered chicken eggshells (CES) as calcium carbonate (CaCO3) bio-filler. These components were incorporated into melamine formaldehyde resin to produce the transparent IFR coating. It was revealed that the developed IFR coatings achieved the highest fire protection rating based on UL94 flammability standards compared to the control. The coatings also yielded increased LOI values, indicative of self-extinguishing properties. A 17 °C elevation of the IFR coating's melting temperature and a significant ∼172% increase in enthalpy change from the control were observed, indicating enhanced fire-retardancy. The thermal stability of the coatings was improved, denoted by reduced mass losses, and increased residual masses after thermal degradation. As validated by microscopy and spectroscopy, the abundance of phosphorus and carbon groups in the coatings' condensed phase after combustion indicates enhanced char formation. In the gas phase, TG-FTIR showed the evolution of non-flammable CO2, and fire-retardant PO and P-O-C. Mechanical property testing confirmed no reduction in the adhesion strength of the IFR coating. With these results, the developed IFR coating exhibited enhanced fire-retardancy whilst remaining optically transparent, suggestive of a dual-phase IFR protective mechanism involving the release of gaseous combustion diluents and the formation of a thermally insulating char layer.

Enamel remineralisation prospect of Moringa Oleifera hydrogel, eggshell hydrogel versus sodium fluoride varnish on artificially demineralised primary teeth: in vitro study.

PURPOSE: The purpose of the present in vitro study is to investigate and compare the remineralising potential of Moringa Oleifera extract, eggshell, and sodium fluoride varnish on microhardness of artificially demineralised enamel of primary teeth with biomimetic minimally invasive approach following the world paradigm shift towards natural products in paediatric dentistry. MATERIAL AND METHODS: Sample size included 44 primary molars. The mineral content and surface microhardness of all specimens were initially assessed using energy dispersive x-ray examination (EDX) and Vickers microhardness. The specimens were artificially demineralised for 96 h at a temperature of 37°C and then reassessed directly after demineralisation. The demineralised enamel specimens were randomly divided into four groups according to the remineralisation regimen utilised. Group 1: Artificial saliva (control); Group 2: Sodium fluoride varnish; Group 3: Eggshell hydrogel; and Group 4: Moringa Oleifera hydrogel. The specimens were stored for 8 days and then subsequently evaluated using EDX and microhardness assessment by Vickers microhardness test and scanning electron microscope (SEM).  Results: Regarding the microhardness test, there was a significant difference between the Moringa Oleifera group and Eggshell group compared to fluoride varnish (p < 0.05). Regarding EDX analysis, there was a statistically significant difference (p < 0.05) between Moringa Oleifera group and Eggshell group compared to fluoride varnish as the highest values were for Moringa Oleifera and Eggshell. On the other hand, there was no statistically significant difference (p > 0.05) between Moringa Oleifera and Eggshell in both the measurements. CONCLUSION: Moringa Oleifera and Eggshell might be considered as a biomimetic natural material capable of guiding enamel tissue remineralisation in early carious lesion of primary teeth. CLINICAL RELEVANCE: This research demonstrated the capability for early enamel caries to be remineralised using novel materials with a naturally counterpart implicated in biomineralisation as proved to be more effective than traditionally used fluoride varnish in primary teeth.

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.

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.

Genomic insights into shank and eggshell color in Italian local chickens.

Eggshell and shank color in poultry is an intriguing topic of research due to the roles in selection, breed recognition, and environmental adaptation. This study delves into the genomics foundations of shank and eggshell pigmentation in Italian local chickens through genome-wide association studies analysis to uncover the mechanisms governing these phenotypes. To this purpose, 483 animals from 20 local breeds (n = 466) and 2 commercial lines (n = 17) were considered and evaluated for shank and eggshell color. All animals were genotyped using the Affymetrix Axiom 600 K Chicken Genotyping Array. As regards shank color, the most interesting locus was detected on chromosome Z, close to the TYRP1 gene, known to play a key role in avian pigmentation. Additionally, several novel loci and genes associated with shank pigmentation, skin pigmentation, UV protection, and melanocyte regulation were identified (e.g., MTAP, CDKN2A, CDKN2B). In eggshell, fewer significant loci were identified, including SLC7A11 and MITF on chromosomes 4 and 12, respectively, associated with melanocyte processes and pigment synthesis. This comprehensive study shed light on the genetic architecture underlying shank and eggshell color in Italian native chicken breeds, contributing to a better understanding of this phenomenon which plays a role in breed identification and conservation, and has ecological and economic implications.

Whole transcriptome sequencing reveals key genes and ceRNA regulatory networks associated with pimpled eggs in hens.

Eggshell is one of the most important indicators of egg quality, and due to low shell strength, pimple eggs (PE) are more susceptible to breakage, thus causing huge economic losses to the egg industry. At the current time, the molecular mechanisms that regulate the formation of pimple eggs are poorly understood. In this study, uterine tissues of PE-laying hens (n = 8) and normal egg (NE) -laying hens (n = 8) were analyzed by whole transcriptome sequencing, and a total of 619 differentially expressed mRNAs (DE mRNAs), 122 differentially expressed lncRNAs (DE lncRNAs) and 21 differentially expressed miRNAs (DE miRNAs) were obtained. Based on the targeting relationship among DE mRNAs, DE lncRNAs and DE miRNAs, we constructed a competitive endogenous RNA (ceRNA) network including 12 DE miRNAs, 19 DE lncRNAs, and 128 DE mRNAs. Considering the large amount of information contained in the network, we constructed a smaller ceRNA network to better understand the complex mechanisms of pimple egg formation. The smaller ceRNA network network contains 7 DE lncRNAs (LOC107056551, LOC121109367, LOC121108909, LOC121108862, LOC112530033, LOC121113165, LOC107054145), 5 DE miRNAs (gga-miR-6568-3p, gga-miR-31-5p, gga-miR-18b-3p, gga-miR-1759-3p, gga-miR-12240-3p) and 7 DE mRNAs (CABP1, DNAJC5, HCN3, HPCA, IBSP, KCNT1, OTOP3), and these differentially expressed genes may play key regulatory roles in the formation of pimpled eggs in hens. This study provides the overall expression profiles of mRNAs, lncRNAs and miRNAs in the uterine tissues of hens, which provides a theoretical basis for further research on the molecular mechanisms of pimpled egg formation, and has potential applications in improving eggshell quality.