Preeclampsia is a pregnancy complication associated with de novo gestational hypertension, proteinuria, and dysregulations in the coagulation and fibrinolytic systems. Platelet-derived factors include various growth and differentiation factors, such as transforming growth factor (TGF)-β. The TGF-β type III receptor (TGFBR3), also known as betaglycan, is a co-receptor for the TGF-β superfamily and an important regulator of reproduction and fetal development. Here, we test the hypothesis whether or not TGFBR3 is involved in platelet-mediated upregulation of placental plasminogen activator inhibitor-1 (PAI-1). Perivillous fibrin deposits and PAI-1 levels were analyzed via immunohistochemistry, gene expression as well as protein expression in healthy and preeclamptic placental tissue. Placental villi and trophoblast cell line BeWo were incubated in presence or absence of washed platelets from pregnant women, platelet-derived soluble factors, or platelet-derived extracellular vesicles. Additionally, BeWo cells were treated with TGF-β-receptor inhibitors and silenced for TGFBR3 expression. PAI-1 and its encoding gene SERPINE1, as well as perivillous fibrin deposits, were significantly increased towards term of gestation and showed a significant upregulation in placental villi from preeclamptic pregnancies. Furthermore, SERPINE1 and PAI-1 were substantially upregulated in response to washed platelets and platelet-derived factors, but not to platelet-derived extracellular vesicles. The upregulation of SERPINE1 was blocked to control levels with the TGF-β inhibitor P144, whereas silencing of TGFBR3 had no effect. Our results suggest that platelet-derived factors induce placental PAI-1 expression independently of TGF-β receptor 3. Increased maternal platelet activation may contribute to significantly upregulated placental PAI-1 and perivillous fibrin deposits in preeclampsia.

Background: Low-dose aspirin (LDA) reduces preeclampsia (PE) risk by up to 40%, yet its molecular effects on chorion trophoblast cells (CTCs), a fetal membrane lineage at the feto-maternal interface, remain obscure. CTCs form a structural and immunoregulatory barrier whose dysfunction drives inflammation-associated membrane pathology in PE. Extracellular vesicles (EVs) released by CTCs may encode cellular stress and adaptation states, offering a molecular window into aspirin's timing-dependent effects on PE risk modification. Methods: Human CTCs were challenged with cigarette smoke extract (CSE) to model oxidative stress-driven PE pathology. Two paradigms were tested: (1) prophylactic aspirin (4 and 40 µg/mL) before and/or flanking the CSE, and (2) therapeutic aspirin after the CSE challenge. The EVs were isolated via ultracentrifugation and size-exclusion chromatography, characterized by nanoparticle tracking and immunoblotting, and profiled by quantitative mass spectrometry. A network pathway analysis and machine learning biomarker selection defined the EV-encoded molecular states. Results: The CTC-derived EVs from the CSE-exposed cells carried a PE-like proteomic signature marked by suppressed VEGF/ECM remodeling, activated TNF-p53 apoptotic signaling, and heightened inflammation. Prophylactic low-dose aspirin shifted the EV cargo toward an EV-encoded signature consistent with preserved angiogenic potential (enrichment of VEGFA, COL1A1, and MMP14) and predicted attenuation of apoptotic and NF-κB pathway activity by an Ingenuity Pathway Analysis. High-dose aspirin produced broad transcriptional suppression without an accompanying pro-angiogenic EV signature. Therapeutic (post-injury) aspirin partially attenuated the injury-associated EV cargo but did not restore the angiogenic EV signature. An exploratory machine learning analysis of EV proteomes identified a candidate prophylactic biomarker panel anchored by HSPA8, SERPINF2, COL4A1, and PLOD1, mapped to the predicted angiogenic recovery and redox-balance pathways. These EV cargo readouts represent the predicted molecular states and require functional validation before clinical interpretation. Conclusions: The CTC-derived EV proteomic signatures capture the dose- and timing-dependent aspirin effects in this in vitro CTC model, positioning the chorion as a candidate pharmacological "secondary responder" favoring cellular resilience over classical anti-inflammatory suppression. As an exploratory hypothesis-generating study, EV-based molecular profiling could provide a foundation for future investigations aimed at stratifying aspirin responders from non-responders, although clinical validation in maternal plasma cohorts will be required before any translational application.

Human follicular development depends on coordinated communication between granulosa cells (GCs) and oocytes through endocrine cues, direct contacts, and extracellular vesicles (EVs). Exosomes are key EV mediators of intrafollicular signaling, but their cargo and functions in gonadotropin-stimulated GCs remain poorly defined. The human granulosa-like tumor cell line KGN was used to investigate exosome secretion and protein composition following human chorionic gonadotropin (hCG) stimulation. Exosomes were isolated by ultracentrifugation, characterized via nanoparticle tracking analysis (NTA), Scanning Electron Microscopy (SEM) and Western blotting, and analyzed using high-resolution mass spectrometry. Comparative proteomics integrating exosomal profiles with the whole secretome were performed, followed by bioinformatic analyses of protein networks, gene ontology, and pathway enrichment. hCG reshaped exosomal cargo, identifying 59 proteins enriched in exosomes, including Integrin α3 (ITGα3), Galectin-3-binding protein (LGALS3BP), tetraspanins (CD63, CD151), and proteasome subunits. Functional enrichment indicated roles in extracellular matrix remodeling, integrin signaling, proteostasis, and steroidogenesis. Comparison with the secretome revealed distinct protein distributions, supporting selective exosomal packaging. Western blot confirmed increased ITGα3 and LGALS3BP levels in exosomes upon hCG treatment. In conclusion, hCG modulates exosome cargo composition in granulosa cells, uncovering a novel mechanism of extracellular regulation.

Successful embryo implantation requires dynamic, bidirectional communication between a developmentally competent blastocyst and a receptive endometrium, integrating hormonal, molecular, and immunologic signals. Increasing evidence indicates that endometrial receptivity is critically dependent on a specialized immune microenvironment that supports trophoblast invasion while maintaining maternal tolerance. This review synthesizes current knowledge on the immunologic regulation of implantation, with emphasis on uterine natural killer (uNK) cells, regulatory T cells (Tregs), macrophages, dendritic cells, and cytokine networks. We further examine intracellular signaling pathways-including JAK/STAT, PI3K/AKT, NF-κB, and MAPK-that integrate immune and decidual responses. The bidirectional embryo-endometrium dialogue is explored through embryo-derived mediators such as human chorionic gonadotropin (hCG), cytokines, growth factors, and extracellular vesicles. The endometrium is increasingly recognized as a biosensor of embryo quality, selectively supporting viable embryos. Disruption of this complex communication network is implicated in recurrent implantation failure and early pregnancy loss. Despite substantial mechanistic advances, clinical translation remains limited. Emerging strategies, including immune profiling, microbiome modulation, and extracellular vesicle-based diagnostics, hold promise for precision reproductive medicine.

The persistent hyperglycemic microenvironment in diabetic wounds causes dysfunction of repair cells, resulting in impaired angiogenesis and disorganized extracellular matrix (ECM) deposition. Small extracellular vesicles (sEVs) derived from hypoxia-challenged chorionic plate mesenchymal stem cells (CP-MSC-sEVsHypo) have been explored as novel acellular therapeutics. However, the effects of oxygen tension during culturing parent cells on the pro-regenerative efficacy and mechanisms of CP-MSC-sEVs have not been systematically investigated. In addition, their rapid clearance and limited interaction with target cells at wound sites constrained therapeutic efficacy. In this study, oxygen tension was first optimized systematically during the precondition of CP-MSCs to improve the pro-regenerative properties of the derived sEVsHypo. Then, sEVsHypo-engineered nanofiber scaffolds were fabricated to achieve the sustained release of sEVsHypo at wound sites by polydopamine (PDA)-mediated interfacial adhesion. In vitro experiments revealed that sEVs obtained under 5% oxygen tension (sEVsHypo-5%) exhibited the most pronounced angiogenic and collagen-regenerative performance. Furthermore, small RNA sequencing and bioinformatic analyses revealed that miR-21-5p was the most abundant miRNA in sEVsHypo-5%, and functional rescue assays validated that miR-21-5p was the key mediator of endothelial activation and ECM remodeling. To improve the bioavailability of sEVsHypo-5%, PDA was leveraged to immobilize sEVsHypo-5% onto a biocompatible, ECM-mimicking poly(ε-caprolactone) nanofiber scaffold to achieve high loading efficacy and sustained release. In a full-thickness diabetic wound model, the sEVsHypo-5%-loaded nanoscaffold accelerated wound closure and achieved superior pro-healing effects on D14 via enhancing neovascularization and ECM deposition, as confirmed by histological and immunofluorescence analyses. Collectively, this sEVsHypo-5%-loaded nanoscaffold achieved efficient immobilization and sustained delivery of sEVsHypo-5% to promote angiogenesis and matrix repair by endogenously delivering miR-21-5p at diabetic wound sites, with strong potential for clinical translation.

The pink bollworm, Pectinophora gossypiella Saunders is a major pest of cotton, notorious for its high reproductive potential and rapid evolution of resistance to Bacillus thuringiensis (Bt) toxins. Despite its economic significance, detailed knowledge of its reproductive anatomy and egg ultrastructure has remained limited, constraining the development of advanced molecular control strategies such as CRISPR/Cas9-based genome editing. The present study provides the first comprehensive characterization of the reproductive system and egg surface morphology of P. gossypiella using stereomicroscopy and scanning electron microscopy (SEM) techniques. The male reproductive system consists of fused, bean-shaped testes, seminal vesicles, duplex and simplex ejaculatory ducts, and paired accessory glands. The female reproductive system comprises paired ovaries with four polytrophic ovarioles per ovary, lateral and common oviducts, accessory glands, corpus bursae, and spermathecal glands. Eggs are oval, dorsoventrally flattened, exhibit a reticulated chorion with distinct micropylar and aeropylar regions. SEM images revealed 6-9 rosette cells encircling a circular micropylar plate, 14-19 first order and 17-23 s order ribs, and 250-291 polygonal surface cells. The structural features of P. gossypiella eggs reveal key sites for sperm entry, aeropylar respiration, and candidate zones for microinjection in gene editing applications. These findings establish a morphological baseline critical for optimizing embryo manipulation and ribonucleoprotein (RNP) delivery in lepidopteran genome editing. This study represents a pioneering effort to integrate classical egg morphology with molecular entomology, thereby advancing precision genetic interventions aimed at resistance management and population suppression in P. gossypiella.

Invasive hydatidiform mole is a rare subtype of gestational trophoblastic disease characterized by myometrial invasion and the potential for significant morbidity. Cesarean scar pregnancy (CSP) is an uncommon type of ectopic pregnancy with increasing incidence due to the rise in cesarean delivery rates. The coexistence of an invasive hydatidiform mole implanted in a cesarean scar is exceptionally rare and poses significant diagnostic and therapeutic challenges. This is a case of a 32-year-old Malay woman with a prior history of cesarean section who presented with per-vaginal bleeding in early pregnancy. Assessment revealed a viable gestational sac with vesicles within, suggesting that a partial hydatidiform mole had implanted at the cesarean scar site. Serum beta-human chorionic gonadotropin (β-hCG) levels were markedly elevated (>200,000 IU/L). MRI of the pelvis was reported as suggestive of partial invasive molar pregnancy with serosal breach through the cesarean scar and bladder invasion, along with a fetus seen, and proceeded with six cycles of methotrexate and folinic acid regimen. Serum β-hCG levels reduced accordingly (368 IU/L pre-sixth cycle and prior surgery). Proceeded with total abdominal hysterectomy, bilateral salpingectomy, and histopathological examination confirmed the diagnosis of an invasive hydatidiform mole arising from a CSP. The patient was managed with a multidisciplinary approach with close β-hCG surveillance. Clinical and biochemical remission was achieved. Invasive hydatidiform mole presenting as a CSP is extremely rare. Early recognition through imaging, β-hCG monitoring, and histopathology is crucial to prevent life-threatening complications. Prompt multidisciplinary management and careful follow-up are essential for favorable outcomes.

The objective of this study was to evaluate the effects of exogenous GnRH administration at the beginning of estrus synchronisation in mares during the spring transitional period. Estrus was synchronised using a progesterone releasing intravaginal device (PRID). The PRID was left in the vagina for 10 days, followed by an injection of 0.4 mg of cloprostenol at PRID removal. The GnRH group (n = 32) was subjected to intramuscular administration of 100 μg of the GnRH agonist triptorelin at PRID insertion, while the control group (n = 32) received 1 mL of sterile physiological saline solution. Ovulation was induced by an intramuscular injection of 3000 IU of human chorionic gonadotropin until the dominant follicle reached a diameter of 35 mm. The mares were examined and insemination was performed. Subsequently, insemination was carried out every 12 h until ovulation. Transrectal palpation and ultrasound were carried out 15 days after ovulation to confirm the presence or absence of an embryonic vesicle. The days of ovulation induction and insemination in the control group were more dispersed than in the GnRH group. Compared with the control group, the time of ovulation induction and insemination in the GnRH group were accelerated and concentrated. In summary, GnRH given at the beginning of the estrus synchronisation program significantly increased synchronisation of ovulation in mares; however, it did not increase pregnancy rates.

Cells can recover lost protein functions through a process we term exofection, in which extracellular vesicles deliver functional molecular cargo to recipient cells and transiently reprogram their activity. Here we show that exosomes derived from fetal chorion trophoblast cells (CTCs) restore P-glycoprotein (P-gp) efflux transporter function in inflammation-impaired maternal decidual cells (DECs) at the feto-maternal interface. CTCs maintain high P-gp expression under inflammatory stress, whereas DECs exhibit marked downregulation of transporter genes and proteins. Proteomic analysis revealed that CTC-derived exosomes package P-gp as a stable cargo that enters DECs through clathrin- and heparan sulfate-dependent uptake pathways. Delivery of CTC exosomes reinstated P-gp abundance and efflux capacity in LPS-stimulated or P-gp-deficient DECs, as shown by calcein efflux and immunofluorescence assays. In pregnant P-gp knockout mice, exosome treatment restored systemic clearance of the P-gp substrate tacrolimus and improved pharmacokinetic parameters. These findings establish exofection as a naturally occurring mechanism of transporter rescue at the feto-maternal interface, where fetal exosomes compensate for inflammation-induced maternal loss of efflux capacity. By restoring P-gp-mediated barrier function, exofection provides a protective strategy that limits the accumulation of xenobiotics and cytokines in maternal tissues and safeguards the fetus. This work reveals a previously unrecognized form of intercellular communication with broad implications for fetal protection, placental biology, cellular engineering, and the delivery of therapeutic proteins.

Trypanosoma cruzi, the causative agent of Chagas disease, can cross the placental barrier and be transmitted congenitally, yet the mechanisms underlying this process remain incompletely understood. Recent evidence suggests that T. cruzi-derived extracellular vesicles (TcEVs) may facilitate placental invasion by modulating host-pathogen interactions. In this study, we examined the effects of TcEVs on human placental explants (HPEs), focusing on their capacity to disrupt tissue architecture and modulate matrix metalloproteinases MMP-2 and MMP-9, enzymes critical for extracellular matrix remodeling. Term placental chorionic villi were cultured ex vivo and exposed to TcEVs, heat-inactivated TcEVs, infective trypomastigotes, or combinations thereof. TcEVs induced ultrastructural damage, including trophoblast detachment and basal lamina disorganization, which were exacerbated by co-incubation with parasites. Immunohistochemistry and Western blotting revealed significant upregulation of MMP-2 and MMP-9, while gelatin zymography confirmed increased enzymatic activity. Our findings demonstrate that TcEVs independently and synergistically with T. cruzi compromise placental integrity by enhancing MMP expression and activity, thereby priming the placental microenvironment for parasite invasion. Targeting TcEVs signaling or MMP activation may represent a novel strategy to prevent congenital transmission of T. cruzi.

Extracellular vesicles (EVs) play a critical role in intercellular communication and have shown great potential in disease treatment and tissue repair. However, large-scale production and clinical application of EVs still face challenges such as donor heterogeneity, batch-to-batch variability, and high production costs. This study aimed to establish a stable and reproducible EV production platform using hTERT-immortalized placental mesenchymal stromal cells (PC-MSCs). We demonstrated that stable expression of the TERT gene promotes cell proliferation in vitro while retaining surface marker expression patterns and differentiation capabilities similar to those of primary cells. The EVs produced by immortalized cells maintain similar particle size distributions, protein markers, and protein expression profiles to those of their parental cells, and their therapeutic efficacy was validated in a bleomycin (BLM)-induced pulmonary fibrosis (PF) rat model. In summary, we systematically described the effect of immortalization via ectopic hTERT gene expression on EVs produced from PC-MSCs. These findings provide a new theoretical basis for the sustainable production and scale-up of EVs, as well as a new solution for the treatment of PF.

Insect egg glue plays a crucial role in adhering eggs together and to the substrate, thus providing them protection and stability. In most insect species, the glue is secreted by the accessory glands in the female reproductive tract, known as 'colleterial glands'. However, in Nezara viridula L. (Hemiptera: Pentatomidae), these glands are absent, leaving its production site unclear. Through light microscopy, Transmission Electron Microscopy (TEM), and Scanning Electron Microscopy (SEM), a distinct glue layer was identified between the follicular cells and the mature eggs in the vitellarium region, extending to the pedicel region of the ovariole. Ultrastructural analysis revealed that follicular cells contained numerous glue-filled vesicles, indicating that they actively secrete the egg glue via microvilli. During oviposition, the barrel-shaped eggs are laid in clutches with glue at their bottom and lateral sides. The glue subsequently hardens to form a cement-like layer that secures the eggs to the substrate. This glue adapts to the surface, faithfully replicating its features. N. viridula eggs show chorion decorations with conical projections and aero-micropylar processes along the circumference of the operculum. Mushroom-shaped structures are present around the aero-micropylar processes, making this area unwettable. The aero-micropylar processes vary in number and have porous texture with the opening for sperm entry in the apical part. This study, together with further investigations aiming to characterize the mechanical and chemical properties of the egg glue of N. viridula, enhances our understanding of the reproductive biology of this harmful insect and can have practical implications for pest control strategies and innovations in materials science.