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.

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.

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.

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).

Fracture repair is a constant clinical challenge, and finding a method to promote and improve restoration is a primary goal for researchers. This is examined from various perspectives, such as fewer complications, increased speed, and cost-effectiveness. The present study aimed to investigate the effectiveness of eggshell powder, compared to the commercial form of demineralized bone matrix (DBM), in critical-size defects in rat calvarial bone. In this study, 40 adult male Wistar rats were selected and randomly divided into four groups of 10. The first group was the control group (C), the second was the eggshell powder group (E), the third was the DBM group (D), and the fourth was the one simultaneously receiving eggshell powder and DBM (DE). In these groups, a 5 mm diameter defect was created in the calvaria using a trephine. All animals received the appropriate treatment for their group. Each group was then divided into two subgroups of five. On days 30 and 60 post-surgery, these subgroups were euthanized, followed by sampling and histopathology examinations. After evaluating the repair percentage using Quick Photo software, the DE group had the highest repair percentage on days 30 and 60. Groups E and D had similar recovery percentages, with group D having a slightly higher one. There was a significant difference between all three groups and the control group. In conclusion, eggshell powder may potentially serve as a suitable substitute for some transplants.

Duck eggs are widely-consumed food and cooking ingredient. The heavier yolk weight (YW) corresponds to a larger size and greater value. However, there is no nondestructive method available to estimate the weight of the yolk. Accurate weight prediction of duck egg yolks must combine both phenotypic and internal information. In this research, we used Visible-Near Infrared (VIS-NIR) spectroscopy to obtain internal information of duck eggs, and a high-definition camera to capture their phenotypic features. YW was predicted by combining the reduced spectral and RGB image information with the whole egg weight. We also investigated the impact of color and thickness of the duck egg on spectral transmittance (ST), as these factors can influence the extent of ST. The results showed that the spectral curves of duck eggs produced 2 peaks and 1 valley, which may be caused by the dual-frequency absorption of the C-H group and O-H group, and can be used to symbolize the internal information of duck eggs. The ST was somewhat affected by the color and thickness of the duck eggshell. Before modelling, Principal component analysis (PCA) was used to significantly reduce the dimensionality of the RGB image with spectral data. A partial least squares regression (PLSR) model was utilized to fit all the features. The test set yielded a coefficient of determination (R2) of 0.82 and a Root Mean Squared Error (RMSE) of 1.05 g. After removing the eggshell\'s color and thickness features, the model showed an R2 of 0.79 and an RMSE of 1.11 g. This study demonstrated that the yolk weight of duck eggs can be estimated using VIS-NIR spectroscopy, RGB images and whole egg weight. Furthermore, the effects of shell color and thickness can be neglected.

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.

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.

OBJECTIVE: 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.
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.
CONCLUSIONS: 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.
CONCLUSIONS: 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.

Calcium and vitamin D deficiencies have been ongoing problems in Koreans due to a lack of food sources of calcium and vitamin D. Postmenopausal women aged 50 to 64 years (n = 25) were randomly assigned to consume three home meal replacements (HMRs)/week with (treatment) and without (control) eggshell powder and vitamin D for 6 months. Additionally, subjects who agreed to continue the study consumed the same three HMRs/week for an additional 6 months in this randomized double-blind study. We confirmed the high compliance of the study participants by analyzing carotenoids, the bioactive substances of HMRs, in the blood. The treatment group consumed an additional 261 mg/d of calcium and 10.3 μg/d of vitamin D from the HMRs, thus meeting the recommended intakes of calcium and vitamin D for Koreans. As a result of consuming fortified HMRs for 6 months, the decline in femoral neck bone density was significantly reduced in the treatment group (p = 0.035). This study indicates that inexpensive eggshell powder may be a good source of calcium for populations with low consumption of milk and dairy products. Additionally, functional HMRs fortified with eggshell powder and vitamin D can be a good dietary strategy for bone health.