Search Results (296)

Objectives: Diminished ovarian reserve (DOR) significantly compromises in vitro fertilization (IVF) success. Although systemic markers such as anti-Müllerian hormone (AMH) serve as valuable clinical indicators of the ovarian reserve, they lack the sensitivity to reflect the qualitative deterioration of the follicular microenvironment. Therefore, in this study, we aimed to characterize the inflammatory proteome of follicular fluid (FF) to establish a high-performance auxiliary diagnostic model for DOR. Methods: Utilizing the ultra-sensitive Olink proximity extension assay, we quantified 92 inflammation-related proteins in the FF of 88 participants (67 with DOR and 21 normal controls). Differentially expressed proteins (DEPs) were identified, and their relationships with key clinical indices were evaluated. A robust predictive signature was refined through integrated Least Absolute Shrinkage and Selection Operator (LASSO) regression and Random Forest algorithms, with diagnostic performance assessed via 10-fold cross-validation. Results: Thirty-five DEPs were significantly dysregulated in the FF of patients with DOR, demonstrating strong associations with serum AMH and basal estradiol concentrations. A minimized diagnostic panel comprising four core proteins, adenosine deaminase (ADA), vascular endothelial growth factor A (VEGFA), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), and matrix metalloproteinase-1 (MMP-1), was established. This multivariable model achieved an excellent area under the receiver operating characteristic curve (AUC) of 0.953. Conclusions: The identified four-protein signature reflects localized chronic inflammation and early pathophysiological shifts in the DOR follicular microenvironment. As a high-performance molecular index, this panel could complement conventional systemic assessments, provide a reliable means of evaluating follicular viability, and optimize individualized therapeutic strategies. Full article

Oocyte maturation is a pivotal event in teleost reproduction that directly determines egg quality, fertilization success, and the developmental competence of early embryos. However, the transcriptional and post-transcriptional regulatory mechanisms operating within oocytes during maturation in marine teleosts remain poorly understood. In the present study, the orange-spotted grouper (Epinephelus coioides), an economically important marine aquaculture species, was used as a model. Oocytes at four distinct maturation stages were obtained by microscopically removing the surrounding follicular layers, followed by integrated mRNA-seq and miRNA-seq analyses to characterize the molecular regulatory landscape underlying oocyte maturation and hydration. The results showed that, as maturation progressed, oocyte diameter and wet weight increased significantly, accompanied by a marked decrease in Na⁺ content, a significant increase in K⁺ content, and the continuous accumulation of most free amino acids, indicating the gradual establishment of an osmotic basis favorable for oocyte hydration. Transcriptomic analysis further revealed extensive transcriptional remodeling during both the early and late phases of maturation. Differentially expressed genes were significantly enriched in pathways related to oocyte meiosis, cytokine signaling, lipid metabolism, DNA replication, cell cycle regulation, ribosome biogenesis, spliceosome function, oxidative phosphorylation, and mitochondrial activity, suggesting that oocyte maturation is a dynamic process characterized by a shift from basal growth maintenance to metabolic reprogramming, maternal transcript remodeling, and terminal maturation responses. miRNA profiling identified a large number of stage-specific differentially expressed miRNAs, including let-7d-5p, miR-22a-3p, and novel-miR-20/27/118, whose predicted target genes were mainly enriched in ribosome-related pathways, oxidative phosphorylation, DNA replication, transcriptional regulation, and signal transduction. Moreover, the miRNA–TF–mRNA regulatory network demonstrated that miRNAs may not only directly repress target genes, but also mediate hierarchical regulatory cascades through transcription factors, thereby coordinately participating in cell cycle progression, cytoskeletal remodeling, vesicular transport, and immune- and cell communication-related responses. Collectively, this study provides the first systematic temporal atlas of mRNA and miRNA regulation during oocyte maturation and hydration at the oocyte level in a marine teleost, thereby deepening our understanding of the molecular basis of meiotic resumption and egg quality formation, and offering valuable theoretical support for the optimization of artificial breeding and the identification of key molecular targets in grouper reproduction. Full article

Ovarian dysfunction resulting from metabolic or toxic injury is characterized by follicular depletion, stromal remodeling, oxidative stress, and endocrine dysregulation. Placenta-derived mesenchymal stem cells (PD-MSCs) have been proposed as a potential therapeutic approach due to their paracrine factors, including placental growth factor (PlGF). However, the pathways through which PD-MSCs exert protective effects on the ovary remain insufficiently defined. In this study, we examined whether PD-MSC transplantation ameliorates ovarian injury in a thioacetamide (TAA)-induced ovarian insufficiency model and explored the signaling events potentially associated with this response. Female rats were administered TAA for 12 weeks, and PD-MSCs were transplanted at week 8. We assessed ovarian morphology, fibrosis, oxidative stress markers, hormonal profiles, and follicle development. Complementary in vitro experiments using TAA-treated KGN granulosa-like cells were performed to investigate potential mechanistic associations. PD-MSC transplantation improved ovarian architecture, reduced collagen deposition, enhanced follicle growth, and mitigated oxidative stress. These changes were accompanied by increased PlGF expression and enhanced activation of fms-like tyrosine kinase-1 (Flt-1), p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), and nuclear factor erythroid 2-related factor 2 (Nrf2)-related antioxidant pathways. In vitro, PD-MSCs coculture similarly attenuated oxidative stress and partially improved mitochondrial membrane potential in damaged KGN cells. Together, these findings suggest that PD-MSCs ameliorate ovarian structural damage and oxidative stress in TAA-induced injury, potentially through paracrine mechanisms partly involving PlGF/Flt-1-associated antioxidant signaling. This work supports the therapeutic potential of PD-MSCs for metabolic or toxicant-induced ovarian insufficiency while underscoring the need for further studies to fully delineate the specific contribution of PlGF and its interaction with downstream antioxidant pathways. Full article

Nerve Growth Factor (NGF), a member of the neurotrophin family, is currently regarded as a key regulator of ovarian physiology beyond its well-known neurotrophic functions. The mammalian ovary is one of the most highly innervated peripheral organs. Increasing evidence indicates that NGF and its receptors, TrkA and p75NTR, are widely expressed in ovarian tissues. Through the activation of the PI3K/AKT, MAPK/ERK, and PLCγ signaling pathways, NGF influences granulosa cell proliferation, steroidogenesis, and ovulation. Physiological levels of NGF are essential for primordial follicle activation, FSH receptor expression, and effective bidirectional communication between oocytes and surrounding somatic cells. As a result, NGF also regulates oocyte maturation and developmental competence. The disruption of NGF signaling can lead to serious health issues. Both low and high levels of NGF negatively affect folliculogenesis and fertility. Elevated intraovarian NGF results in sympathetic over-innervation, altered steroid production, and polycystic ovarian features. In addition, increased NGF expression has been linked to endometriosis and ovarian cancer progression. Clinical studies further suggest that follicular NGF levels may serve as indicators of ovarian reserve and reproductive outcomes in assisted reproduction. This narrative review synthesizes the current knowledge on NGF roles in ovarian physiology and disease. It highlights NGF’ dual functions as a central regulator of follicular dynamics, and as a potential biomarker and therapeutic target for common reproductive system diseases. Full article

Ovarian tissue cryopreservation is the primary fertility preservation strategy for prepubertal girls and patients requiring urgent gonadotoxic therapy. However, the risk of reintroducing malignant cells has prompted the development of safer alternatives, including follicle isolation followed by three-dimensional scaffold encapsulation for transplantation. Fibrin is a promising biomaterial for bioengineered ovary construction, although its ability to support early human follicle maintenance remains unclear. Follicles isolated from cryopreserved ovarian tissues of six patients were encapsulated within fibrin scaffolds of graded concentrations (high, medium, low). After 7 days of in vitro culture, follicle survival and diameter change were quantified. A total of 282 follicles (45.4 ± 10.1 µm) were embedded into fibrin scaffolds. After culture, 237 viable follicles were detected, yielding an overall survival of 84%. Follicle diameter increased to 58.8 ± 12.0 µm. Follicle survival rates were comparable across groups, while mean follicle diameter was 56.3 ± 12.5 µm (high), 61.9 ± 13.4 µm (medium), and 57.4 ± 9.3 µm (low). Follicles cultured in medium-concentration fibrin demonstrated significantly larger diameters compared with both high and low groups (p < 0.05), with no difference between high and low groups. Fibrin-based bioprosthetic ovary scaffolds support short-term in vitro maintenance of isolated human follicles, preserving spherical morphology and granulosa cell layer integrity. Medium-concentration fibrin was associated with greater follicle diameter expansion compared with higher and lower concentrations, indicating that scaffold composition influences early morphometric changes during in vitro follicle culture. Full article

Dermal papilla (DP) cells orchestrate hair follicle growth and cycling by secreting signaling molecules that stimulate follicular epithelial stem cells. The signal peptide CUB-EGF-like domain-containing protein 3 (SCUBE3) was recently identified as a potent anagen stimulator secreted by DP cells. Ageratum houstonianum ethanolic extract (AHE) and its active constituent agerarin exhibit anti-inflammatory properties; however, their effects on hair follicle growth remain unclear. This study aimed to investigate the effects of AHE and agerarin on SCUBE3 expression in primary human DP cells and to elucidate the underlying molecular signaling pathway. Cell viability was assessed by measuring cell confluency. Ex vivo hair growth was analyzed using organ cultures of human hair follicles. Gene and protein expression were determined using reverse transcription-PCR, immunoblot analysis, immunofluorescent staining, tyramide signal amplification-based multiplex immunohistochemistry, and gene promoter-reporter assay in primary human follicle DP cells. In a hair follicle organ culture model, both AHE and agerarin increased the population of the anagen phase and promoted hair shaft elongation. AHE and agerarin significantly upregulated SCUBE3 expression at both the mRNA and protein levels. Mechanistically, AHE and agerarin induced activator protein-1 (AP-1) expression by activating mitogen-activated protein kinase signaling pathways, thereby increasing SCUBE3 gene promoter activity. AHE and agerarin promoted hair follicle growth by upregulating SCUBE3 expression via activation of the MAPK–AP-1 signaling axis. In conclusion, AHE and agerarin may serve as potential therapeutic agents for the prevention and treatment of alopecia (hair loss). Full article

Human hair performs a number of important physiological and esthetic functions. Hair loss and alopecia are complex disorders which affect people all over the world. Hair loss can be an early manifestation of various autoimmunological disorders. Despite a growing interest of researchers in the role of immune factors—especially autoantibodies—in the etiology of certain types of alopecia, their role in alopecia remains uncertain. Several potential autoantigens of follicular components, mainly derived from keratinocytes and melanocytes of the hair follicles, have been found to play a role in the development of alopecia areata. The list of autoantigens includes trichohyalin, keratin 16, fibroblast growth factor receptor 3, glycoprotein-100, melanoma-associated antigen recognized by T cells 1, dopachrome tautomerase/tyrosinase-related protein 2, tyrosinase, and tyrosine hydroxylase. This narrative review presents different aspects of immunopathogenesis of alopecia, from physiology (hair follicle immune privilege) to pathology (disruption of hair follicle immune privilege) and signaling pathways. Identification of key autoantigens could potentially pave the way for the development of new, effective, and more targeted immunotherapies for alopecia. Full article

Bidirectional communication between the oocyte and surrounding follicular cells coordinates follicle growth, meiotic maturation, and the acquisition of competence. We aimed to identify genes related to follicular crosstalk and the secretory pathway as candidate mediators of cumulus–oocyte complex (COC) crosstalk in cattle. Expressed sequence tags (ESTs) from bovine COCs were retrieved from databases and screened for genes related to secretion and the secretory pathway using SignalP and SecretomeP, and transmembrane proteins were removed, yielding 13 candidate genes. Candidate expression was examined in two GEO RNA-seq datasets to assess enrichment in oocytes versus cumulus cells. RT–qPCR profiling across tissues and reproductive cell types enabled principal component analysis and correlation/network analysis, visualized as heatmaps and Cytoscape, revealing an INBHA-SERPINE2-SDF2L1 co-expression pattern. INHBA and SERPINE2 protein products are secreted, whereas SDF2L1 protein is a secretory pathway-associated, endoplasmic reticulum-resident chaperone. Promoter sequences of INHBA, SERPINE2, and SDF2L1 were scanned with FIMO using JASPAR motifs, identifying shared SMAD-associated motifs and FSH/cAMP-related motif families. The data support a co-regulation model in which endocrine FSH/cAMP and activin/TGF-β–SMAD inputs converge on a shared transcriptional program consistent with a putative INHBA–SERPINE2–SDF2L1 co-regulated module, linking cumulus extracellular matrix remodeling/protease control with oocyte ER protein folding capacity during COC maturation. Full article

Skin and hair follicles regenerate through coordinated stem cell niches and cyclic signaling associated with transitions among anagen, catagen, and telogen phases. In alopecia and chronic skin diseases, follicular miniaturization, immune dysregulation, persistent inflammation, impaired vascularization, and a compromised stratum corneum barrier limit the effectiveness of conventional topical and systemic therapies. Bio-nanovesicles (BNVs), including natural extracellular vesicles such as exosomes and microvesicles, as well as engineered artificial or hybrid nanovesicles, offer a targeted, cell-free delivery platform for miRNAs, proteins, and growth factors. By modulating key pathways—Wnt/β-catenin, PI3K/AKT, MAPK/ERK, and TGF-β/BMP—BNVs have the potential to restore regenerative crosstalk, enhance angiogenesis, and help initiate hair and skin repair. Full article

Hair loss affects over half of adults by age 70 and represents a major determinant of overall hair health, imposing significant psychosocial burden across genders. Nutritional factors play a critical role in follicle biology, yet targeted strategies remain underexplored. This comprehensive review examines five key hair-constituent macromolecules—type I collagen, elastin, keratin, ceramides, and melanin—and their physiological and clinical impacts on hair structure, density, shining, and growth. We conducted a structured literature search in PubMed and Google Scholar through January 2025, selecting in vitro studies, animal experiments, and human clinical trials that evaluated each macromolecule’s effects on follicular function and hair fiber integrity. Type I collagen enhances dermal papilla cell proliferation, upregulates Wnt/β-catenin and growth factors, and improves hair thickness and breakage resistance in randomized controlled trials. Keratin hydrolysates replenish cortical protein, reinforce disulfide cross-links, and reduce telogen shedding, with clinical studies demonstrating 30–50% decreases in hair loss and gains in tensile strength. Oral ceramide formulations restore the cuticular lipid barrier, shift follicles toward anagen, and increase hair density in double-blind trials. Although direct clinical data on melanin supplementation are lacking, ex vivo and animal models confirm its role as a UV-protective pigment, preserving keratin integrity and color fastness. Together, these macromolecules constitute a coherent framework for hair health, and clinical studies increasingly provide evidence that their combined or parallel application can meaningfully enhance hair density, strength, shine, and resilience. Full article

Reproductive efficiency in cattle is critically dependent on embryo quantity and quality, particularly in assisted reproductive technology (ART) programs such as superovulation, embryo transfer, and embryo production. Nutrition is a key determinant of embryo yield through its regulatory effects on metabolic signaling, ovarian function, oocyte competence, and early embryogenesis. This review synthesizes the current evidence describing mechanistic links between nutritional status and embryo production in dairy and beef cattle across both in vivo and in vitro systems. Energy balance, protein supply, micronutrients, and fatty acids influence metabolic hormones including insulin, insulin-like growth factor-1, and leptin, which regulate hypothalamic–pituitary–gonadal axis activity, follicular recruitment, and steroidogenesis. Negative energy balance disrupts endocrine signaling, elevates circulating non-esterified fatty acids, increases oxidative stress, and impairs oocyte mitochondrial function, resulting in reduced embryo yield, compromised blastocyst quality, and diminished cryotolerance. Targeted micronutrients such as selenium, zinc, vitamins A and E, B-complex vitamins, and omega-3 fatty acids enhance antioxidant capacity, membrane integrity, and epigenetic regulation, thereby supporting embryo viability and post-transfer survival. Furthermore, early-life nutrition programs long-term reproductive capacity by influencing ovarian reserve establishment and oocyte epigenetic competence. Strategic nutritional management is therefore essential to optimize ART outcomes and promote sustainable genetic progress in cattle production systems. Full article

Goat reproductive performance is a key determinant of the productivity and economic value of goat farming, especially in meat and milk production. In a previous study, to investigate the genetic basis of prolificacy, we divided goats into groups according to their consistent reproductive performance (producing either single kids or twins) over five consecutive kidding cycles, and performed whole-genome resequencing and RNA-seq analysis on their ovarian tissues. Through integrated analysis, we identified three candidate genes—IGF2BP1 (insulin-like growth factor 2 mRNA-binding protein 1), CDC25A (cell division cycle 25A), and RXFP2 (relaxin family peptide receptor 2)—as potential key regulators of reproductive capacity. Using goat ovarian granulosa cells, we systematically assessed the impact of each gene through gain- and loss-of-function experiments. Overexpression of IGF2BP1 promoted cell proliferation and suppressed apoptosis, underscoring its role in maintaining cellular viability. Conversely, its knockdown significantly impeded growth and induced cell death. Similarly, CDC25A enhanced granulosa cell proliferation, whereas its knockdown led to marked growth impairment and increased apoptosis. Proliferation was also enhanced by RXFP2 overexpression but impaired upon its knockdown, suggesting that RXFP2 is functionally important for follicular development. Collectively, these findings establish IGF2BP1, CDC25A, and RXFP2 as fundamental regulators of granulosa cell dynamics and ovarian follicular development, providing crucial functional insights and promising targets for genetic selection to enhance reproductive efficiency in goats. Full article

Background/Objectives: Dynamic remodeling of the zona pellucida (ZP) is a fundamental biochemical and structural process during human preimplantation development; however, its quantitative characterization and clinical relevance remain incompletely defined. The objective of this study was to evaluate dynamic ZP thinning as a functional marker of embryo developmental competence and to examine its relationship with follicular fluid (FF) biomarkers and clinical pregnancy. Methods: This prospective observational study included 47 IVF cycles performed at a single center, yielding 64 transferred blastocysts with complete time-lapse data. ZP thickness was measured from fertilization to 120 h post-fertilization using time-lapse imaging. Two quantitative parameters were derived: the relative thinning ratio (Δrel) and the linear thinning rate (slope). FF concentrations of growth differentiation factor 9 (GDF-9), hyaluronic acid (HA), and syndecan-4 (Syn4) were quantified by ELISA. Embryo-level associations with spontaneous blastocyst hatching were assessed using logistic regression and multivariate analyses, while patient-level models evaluated predictors of clinical pregnancy. Results: Embryos that underwent spontaneous hatching exhibited significantly greater Δrel than non-hatching embryos (p < 0.001). Δrel remained the strongest predictor of hatching in multivariable models (AUC = 0.91). Among FF biomarkers, only GDF-9 showed a positive association with spontaneous hatching. At the patient level, higher Δrel values of transferred embryos were associated with clinical pregnancy (OR 3.65, p = 0.009), whereas FF biomarkers and assisted hatching showed no significant association. Conclusions: Dynamic ZP thinning quantified by Δrel represents a promising indicator of embryo developmental competence. The concordance between embryo-level hatching behavior and patient-level clinical pregnancy suggests potential clinical relevance of ZP dynamics as an integrative embryological marker, warranting validation in larger cohorts. Full article

Hair aging, a complex physiological process involving progressive hair thinning and loss of luster, is primarily driven by functional decline of hair follicle components and sebaceous glands due to cumulative oxidative stress. This decline manifests as dermal papilla cell (DPC) senescence, with reduced insulin-like growth factor-1 (IGF-1) secretion, impaired hair matrix keratinocyte proliferation, and decreased keratin synthesis. We investigated the restorative potential of poly-D,L-lactic acid (PDLLA) filler, a biostimulatory polymer with antioxidant properties, against these age-related changes. PDLLA filler treatment significantly reduced oxidative stress—as indicated by decreased 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels—in hydrogen peroxide-induced senescent human DPCs, alleviated cell-cycle arrest, and significantly upregulated IGF-1 secretion. Conditioned medium from PDLLA filler-treated DPCs stimulated proliferation and pan-keratin expression in senescent hair follicular keratinocytes (HFKs). Intradermal PDLLA filler injection in aged mice significantly reduced 8-OHdG levels, restored DPC proliferative capacity (indicated by proliferating cell nuclear antigen [PCNA] positivity), increased IGF-1 expression within the dermal papilla, and enhanced HFK proliferation in the hair matrix. Consequently, PDLLA filler treatment robustly upregulated hair cortex keratins (K35, K85) and inner root sheath markers (AE15, K25, K71), leading to improved cuticle integrity and the attenuation of follicular miniaturization. Senescence within sebaceous glands was also mitigated, as evidenced by increased PCNA and peroxisome proliferator-activated receptor gamma (PPAR-γ) expression, accompanied by enhanced hair shaft reflectivity and shine. Overall, PDLLA filler ameliorated senescence-associated phenotypes and restored senescence-associated functional decline, supporting its potential as an intervention for age-related hair thinning and quality deterioration. Full article

Background/Objectives: Lifestyle interventions are first-line treatment for women with polycystic ovary syndrome (PCOS) to improve metabolic health. Impacts on reproductive function are less clear. Previous research has been limited by inconsistencies in evaluation of ovulatory function and lack of comparisons between women with and without PCOS. Methods: The present study implemented a prospective clinical trial of 28 women (PCOS, N = 10 and Non-PCOS Control, N = 18) undergoing a 1-month baseline assessment followed by a 6-month hypocaloric dietary intervention. Results: Both groups reached clinically meaningful weight loss with the intervention (PCOS group: 6.5 ± 5.5%; Non-PCOS Control group: 10.0 ± 4.7%). Largest follicle diameter and growth rate of ovulatory dominant follicle, menstrual cycle length and luteal phase length did not change during the intervention in either group (all p > 0.05). The Non-PCOS Control group had increased mid-luteal phase progesterone levels and secretory phase maximum endometrial thickness during the intervention (all p < 0.05), whereas the PCOS group showed a reduction in follicular phase length (p < 0.05). Additionally, changes in ovulatory function and endometrial development were not associated with the rate of weight loss (all p > 0.05). Conclusions: This study demonstrates that women with PCOS are unlikely to experience changes in menstrual cyclicity and endometrial development with a short-term hypocaloric dietary intervention. The shortening of the follicular phase suggests that women with PCOS may need a longer intervention to achieve clinically meaningful improvements in ovulatory function and endometrial health. Full article