Uterine luminal fluid influences embryonic development and the subsequent phenotype of offspring, yet its detailed metabolomic composition remains poorly characterized. Here, minimally invasive transcervical techniques were employed to collect neat uterine fluid from postpartum dairy cows and cyclic beef cows to allow for metabolomic profiling via targeted mass spectrometry. Objectives were to 1) compare the metabolomic profile of uterine fluid with plasma in dairy cows and 2) assess the impact of dietary rumen-protected methionine and stage of estrous cycle (day 0 vs 7) on plasma and uterine fluid metabolomic profile in beef cows. Results revealed that the concentrations of many metabolites, including amino acids, signaling molecules (e.g. dopamine, gamma-aminobutyric acid) and lipids (e.g. ceramides, diacylglycerols), were higher in uterine fluid than in plasma. An oral bolus of rumen-protected methionine increased uterine concentration of methionine on day 0 of the estrous cycle. The uterine metabolome remained relatively stable between days 0 and 7 although there was temporal variability for a select number of metabolites (cysteine, methionine, methionine sulfoxide, asymmetric dimethylarginine, ceramides, and glycerophospholipids). Correlations between plasma and uterine fluid concentrations were strong or moderate for many amino acids. Collectively, these findings highlight that the uterine lumen is a specialized, selectively regulated biochemical compartment.

Objective: Estrogen receptor α (ERα/ Esr1 ) is essential for uterine function during early pregnancy. The uterus has a dynamic immune environment, including transient endometrial inflammation in response to post-coital semen. The uterine epithelium is the first contact for uterine lumen contents (e.g., semen). We aimed to determine the function and mechanism of uterine epithelial ERα in regulating the uterine immune response to semen. Methods: Uterine tissues and uterine flushes were collected from naturally mated uterine epithelial ERα-deficient epiERα -/- ( Esr1 f/- Wnt7a Cre/+ ) mice and Esr1 f/- control mice on day 0.5 post-coitus (D0.5) and D3.5. Histology, immunohistochemistry, mRNA-seq, real-time PCR, flow cytometry, and cytokine assays were employed to determine spatiotemporal distribution of immune cells and assess their functional states. Results: There was a sharp decline in neutrophils from D0.5 to D3.5 in both Esr1 f/- and epiERα -/- uteri. Immunohistochemistry detected ~7-fold ELANE+ neutrophils in uterine luminal epithelium (LE) layer and ~1.5-fold in stromal layer of D0.5 epiERα -/- uterus compared to D0.5 Esr1 f/- counterparts. Gelled uterine lumen contents with varied densities of neutrophils and sperm were detected in both D0.5 Esr1 f/- and epiERα -/- mice. Flow cytometry revealed significantly increased percentages of neutrophils among CD45+ leukocytes in uterine digests with a trend of increased neutrophil cell number and proportions in uterine flushes from D0.5 epiERα -/- mice. Dysregulation of immune genes at both mRNA and protein levels was noted in D0.5 epiERα -/- LE/uterus, especially an upregulation of inflammatory cytokines. In particular, upregulation of genes involved in signaling by interleukin (IL)-1β, a potent pro-inflammatory cytokine involved in mating-induced uterine inflammation, was observed in both the uterine tissue and uterine flush. Conclusion: These findings demonstrate an essential function of uterine epithelial ERα in controlling mating-induced inflammation in the LE layer, stromal layer, and uterine lumen, and highlight the IL-1β signaling pathway among the molecular mechanisms involved in regulating uterine inflammation.

Pregnancy establishment in mammals requires a complex sequence of events, including bi-lateral embryo-maternal communication, leading up to implantation. This is the time when most pregnancy loss occurs in mammals (including humans and food production species) and dysregulation in embryo-maternal communication contributes to pregnancy loss. Embryo-derived factors modify the function of the endometrium for pregnancy success. We hypothesise that these previously unexplored conceptus-derived proteins may be involved in altering the function of the endometrium to facilitate early pregnancy events in mammals with different early pregnancy phenotypes. Here, we show that protein disulphide-isomerase (PDI) is a highly conserved protein among mammals and provide evidence for a species-specific role for PDI in endometrial function in mammals with different implantation strategies. We show how PDI alters the endometrial transcriptome in human and bovine in vitro in a species-specific manner and using a microfluidic approach we demonstrate that it alters the secretome capability of the endometrium. We also provide evidence from in vitro assays using human-derived cells that MNS1, a transcript commonly downregulated in response to PDI in human and bovine endometrial epithelial cells, may be involved in the attachment phase of implantation. We propose that the trophoblast-derived protein PDI, is involved in supporting the modulation of the uterine luminal fluid secreted by the endometrium to support conceptus nourishment, and in the process of embryo attachment to the uterine lumen for pregnancy success in mammals.

Evaluating differences in uterine microbiome and inflammatory status at 1 month postpartum associated with metritis and antibiotic treatment.

In manmalian early pregnancy, maternal immune cells are critical during early pregnancy for initiating embryogenesis and maintaining implantation. However, the preimplantation immune environment within the reproductive tract is largely uncharacterized. Here, we detected a massive influx of immune cells into the lumen of the oviduct and uterusafter mating in mice. In immune cells, CD11b+ cells up to about 38.1% in the oviduct and 59.5% in the uterus, of which neutrophils accounted for 81.7% and 97.7%, respectively. Neutrophils presented a antigen-presenting cells like phenotype (nAPCs), a specialized population expressing Ly6G and MHCII. We followed the dynamic changes and fate of neutrophils, and found that these cells rapidly disappeared from the lumen of the reproductive tract from 0.5 days post-coitus (dpc) to 1.5 dpc. Instead of apoptosis, they migrated from the oviduct to the uterus and infiltrated the endometrium at 1.0-1.5 dpc. At 3.5 dpc, the number of immune cells decreased and natural killer cells became the predominant population. The cytokines involved in modulating immune cell responses (GM-CSF, IFN-γ and IL17A), pro-inflammation (IL-1β, IL-6, IL-12p70, TNF-α), and anti-inflammation (IL-10) were detected in luminal fluid, in which GM-CSF and IL-6 decreased, IL-10 increased in 3.5 dpc uterus. However, spontaneous abortion mice (CBA/J×DBA/2) exhibited altered immune patterns at the stage of embryogenesis and implantation. Our findings reveal a dynamic and finely tuned immune state in the lumen of the oviduct and uterus, which provide a microenvironment for early embryo development, and provide insights into pre-implantation embryo-uterine cross-talk in mice.

Endometrial organoids (EOs) have gained attention as a promising in vitro model for investigating uterine physiology, reproductive disorders, and embryo-maternal interactions, providing an alternative to in vivo studies while minimizing ethical concerns. Despite their increasing use across species, a well-characterized rat EO model is limited. We established and validated a rat EO platform that recapitulates the structural and functional characteristics of the native endometrium. We established and validated a rat EO platform that recapitulates the structural and functional characteristics of the native endometrium. Organoids were generated from epithelial-rich stem-cell populations isolated from adult female rats and cultured in 3D Matrigel. EO formation efficiency was assessed in relation to plasma progesterone concentration, and organoids were evaluated for long-term viability, cryopreservation tolerance, and morphological consistency over serial passages. Functional relevance was examined by real-time polymerase chain reaction and RNA sequencing of sex steroid hormone receptors (progesterone receptor and estrogen receptor α) and CD34. GFP (Green Fluorescent Protein)-labeled EOs were transplanted into the uterine lumen of wild-type rats to evaluate engraftment and persistence. Rat EOs displayed morphological and molecular characteristics comparable to native uterine tissue, maintaining viability and integrity over multiple passages and after cryopreservation. Immunohistochemical analyses using epithelial (E-cadherin), stromal (Vimentin), and proliferative (Ki-67) markers confirmed the presence of multiple cell types resembling those in native uterine tissue. Formation efficiency positively correlated with circulating progesterone concentrations. Gene expression confirmed key endometrial markers, including hormone receptors and stromal-associated genes. GFP-expressing EOs successfully engrafted into wild-type uterine lumens and persisted long term, demonstrating functional and structural compatibility with the in vivo uterine environment. The rat EO model developed here provides a physiologically relevant platform for studying endometrial biology, enabling research on reproductive mechanisms and disease modeling. Its ability to mimic and engraft in the uterine environment suggests applications in regenerative medicine and therapeutic transplantation. This rat EO model provides a physiologically relevant platform for studying uterine biology and reproductive mechanisms without extensive animal use. Its ability to mimic and engraft in the uterine environment supports potential applications in disease modeling, drug testing, and regenerative medicine.

Ultrasonography is a useful non-invasive technique to measure the foetal growth, age and viability in goats. The maternal and foetal parameters such as uterine lumen diameter, placentome size, crown rump length, biparietal diameter, foetal orbit length, trunk diameter, occipito-snout diameter, thoracic diameter, long and short axis of heart, diameter of aorta, umbilical cord diameter, humerus, femoral and tibial length were studied in different breeds of goats to assess the foetal development. Doppler ultrasonic technique has been used to monitor the uterine and umbilical arterial blood flow in pregnant animals. However, there is a scarcity of data on the haemodynamic indices and blood flow velocity of the uterine artery during pregnancy to determine the normal physiological or pathological states. These parameters could serve as valuable determinants for the normal intrauterine foetal development and thus differentiates from gestational pathologies in goats. This review highlights the pivotal role of ultrasonography and Doppler techniques in advancing our understanding of foetal growth and maternal hemodynamics in goats.

Abstract In cattle, during the second week of pregnancy, the embryo transitions from the spherical, zona-enclosed blastocyst to the initially elongated conceptus. The overarching hypothesis is that endometrial function changes temporally to support embryo development. The experimental hypothesis is that concentrations of amino acids (AA) and lipids in the uterine luminal mucus (ULM), and the gene expression of luminal epithelial cells (LE) change between day 7 (D7) and D14 of the estrous cycle. Objective was to compare the concentrations of AA and lipids in the ULM, and the expression of target genes in LE among D7, D10 and D14, which correspond to the blastocyst stage, hatching from the zona pellucida and conceptus initial elongation, respectively. ULM and LE were collected from the uterine body of primiparous, non-lactating, cyclic, Bos indicus-influenced crossbred cows (n=5) using a cytology brush in each of D7, D10 and D14 after synchronized estrus. Targeted metabolomics of ULM was performed by mass spectrometry and gene expression of LE was determined by RNA-seq. Univariate analysis of select metabolites and expression of select genes was conducted to evaluate the effect of day. The concentration of five essential (Arg, Gln, His, Trp and Val) and six non-essential (Ala, Asn, Asp, Glu, Gly and Ser) AA changed over time (P≤ 0.1), out of the 17 AA detected in the ULM. In general, AA concentrations increased from D7 to D10 and decreased from D10 to D14. A list was generated with 152 genes expressed in the bovine LE that potentially participate in synthesis, degradation and transport of AA in ULM, that were affected by time (P ≤ 0.1). There were 37 genes associated with synthesis of non-essential AA (e.g., PHGDH, PSPH, ADSS2), 43 genes associated with catabolism (e.g., FAH, ASRGL1, HPD) and 12 genes associated with transport of AA (e.g. SLC36A2, SLC6A14, SLC6A9). The expression of 36 genes increased from D7 to D10 and then remained constant, and that of 47 genes decreased from D7 to D10 then remained constant. The concentration of three diacylglycerols, 33 glycerophospholipids, five glycosylceramides, six sphingolipids and nine triacylglycerols out of 25, 60, 32, 13 and 24 detected in ULM, respectively, changed overtime (P ≤ 0.1). In general, there was a gradual increase in the concentration of lipids in the ULM from D7 to D14. Analysis of the expression of genes potentially involved in the regulation of lipid concentrations in the ULM is currently underway. In conclusion, the luminal metabolome and gene expression changed dramatically during the second week of the estrous cycle, associated with the blastocyst-to-elongation transition. Disruption in these dynamics may influence embryonic development and pregnancy outcomes in cattle. This research was supported by USDA NIFA Award 2022-67015-36839.

Mammals exhibit unique and highly variable mechanisms for the establishment and maintenance of pregnancy. Ruminants (e.g., sheep, cows, and goats) have novel mechanisms whereby the conceptus (embryo and its extra-embryonic membranes) signals for the establishment of pregnancy and exhibits unique metabolic pathways favoring conceptus development. Embryos of ruminants reach the spherical blastocyst stage at 5 to 10 mm in diameter and then elongate rapidly to elongated filamentous conceptuses of greater than 250 mm as they make contact with the uterine luminal epithelium (LE) for implantation. During conceptus elongation the trophectoderm cells secrete interferon tau (IFNT), a novel pregnancy recognition signal for ruminants to ensure maintenance of a functional corpus luteum (CL) to secrete progesterone (P4) required for pregnancy. P4 induces uterine epithelia cells to express the endogenous Jaagsiekte Retrovirus (enJSRV) that may transactivate toll-like receptors 7 and 8 in the conceptus trophectoderm to induce secretion of IFNT, a classical viral-antiviral mechanism. IFNT silences expression of receptors for estradiol (E2) and oxytocin (OXTR), which abrogates the mechanism whereby oxytocin from CL and posterior pituitary would otherwise induce large pulses of prostaglandin F2α (PGF) by uterine epithelia to cause regression of the CL and its secretion of P4. IFNT has another novel role in silencing expression of not only ESR1 and OXTR, but all classical interferon-stimulated genes in the uterine LE and superficial glandular epithelium (sGE), but with P4 increasing expression of genes for transport of nutrients such as glucose and arginine into the uterine lumen to support conceptus development. Ruminant conceptuses convert glucose to fructose, a novel hexose sugar that cannot be transported back to the maternal circulation. Fructose is converted to fructose-1-PO4 for metabolism, not via the pathway for glycolysis but via the novel fructolysis pathway uninhibited by low pH, citrate, or ATP as is the case for glycolysis. Thus, fructose and its metabolites support the pentose cycle, hexosamine biosynthesis pathway, one-carbon metabolism, and the citric acid cycle for all cells of the conceptus. Arginine is another key nutrient transported into the uterine lumen by the uterine LE/sGE in response to P4 and IFNT. Arginine is metabolized to generate nitric oxide, polyamines, and creatine, essential for conceptus growth and development, while enhancing production of IFNT as a novel pregnancy recognition signal, and upregulating expression of genes in the uterine LE/sGE for transport of nutrients. Fructose is the major hexose sugar supporting major metabolic pathways required for conceptus growth and development in ruminants.

The peri-implantation period of pregnancy in pigs is characterized by rapid morphological transitions of the conceptus necessitating a precisely regulated uterine environment to support elongation, survival, and implantation. Uterine histotroph, composed of nutrients and signaling molecules secreted by or transported by endometrial epithelia, plays a central role in mediating these events. However, dynamic changes in the metabolic composition of uterine luminal fluid (ULF) during early pregnancy are incompletely defined. In this study, we performed stage-resolved, untargeted metabolomic profiling of ULF collected from cyclic and pregnant gilts on Days 10, 12, 14, and 16 of the estrous cycle and pregnancy (n = 2–6/group). A total of 206 metabolites were identified, with amino acids, fatty acids, and carbohydrates being the dominant classes. Principal component and supervised learning analyses revealed progressive divergence in ULF composition between pregnant and cyclic gilts with the most distinct profiles observed by Day 16. Notably, pregnancy induced substantial increases in amino acids associated with mechanistic target of rapamycin (mTOR) signaling and trophectoderm proliferation, including arginine, glutamine, proline, lysine, and phenylalanine. Kyoto Encyclopedia of Genes and Genomes enrichment analyses identified gestational age-dependent activation of pathways involved in amino acid biosynthesis, nucleotide metabolism, and phospholipid turnover. Metabolites such as phosphorylcholine, succinic acid, and asymmetric dimethylarginine increased markedly in pregnancy, suggesting coordinated regulation of membrane remodeling, energy production, and nitric oxide signaling. Targeted quantification of 19 amino acids revealed both linear and quadratic trends across time and pregnancy status, with distinct differences in glycine and serine trajectories between pregnant and cyclic ULF. Collectively, these findings describe the evolving biochemical landscape of the uterine lumen during early pregnancy and highlight key metabolic pathways that likely support conceptus development and uterine receptivity to implantation.

Pyometra is one of the most common pathologies affecting the reproductive system of females, including canines. This condition is characterized by mucopurulent secretion in the uterine lumen and can be classified into open- and closed-cervix pyometra. This pathology is often seen in middle-aged to elderly female dogs because of increased exposure to progesterone and in animals subjected to the use of exogenous progestogens. The most common clinical signs are mucopurulent or bloody discharge from the vulva, swollen vulva, fever, vomiting, anorexia, lethargy, polyuria, polydipsia, weight loss, and prostration. Pyometra may result in uterine enlargement, torsion, rupture, and septicemia. This report describes the case of a young female dog with open pyometra, which was 1 year and 22 days old and had not been exposed to exogenous progestogens. This fact highlights the need to understand the pathology as timely diagnosis aids in rapid and effective treatment.

Histotroph is regulated by P4 and paracrine factors and contains amino acids essential for porcine conceptus development. We hypothesized P4 contributes to amino acid transport during pregnancy in pigs. Ovariectomized gilts received daily P4 or corn oil (CO) injections from Days 12 through 39 post-estrus. High-performance liquid chromatography (HPLC) determined increased abundances of amino acids including glycine, in uterine flushings from P4-treated gilts. SLCs transport amino acids across membranes. rtPCR analyses revealed endometrial expression of SLC6A9 mRNA increased during the peri-implantation period of pregnancy. SLC7A8 and SLC38A1 mRNA expression also increased during that period but decreased later in pregnancy. SLC1A4, SLC1A5, and SLC7A10 mRNA expression was greatest later in pregnancy. Immunofluorescence analyses localized SLC6A9, which transports glycine exclusively, to the endometrial luminal epithelium from Day 12 through 25, uterine glandular epithelium throughout gestation, conceptus trophectoderm and endoderm between Days 15 and 20, and chorionic epithelia (CE) of interareolar and areolar CE between Days 40 and 60. Expression in interareolar regions decreased by Day 60 but remained high in the areolae. Expression of SLC6A9 was greater for endometrium from P4-treated than CO-treated pigs. These results illustrate the complexity of, and stage-dependent changes in, expression of amino acid transporters in endometria from pregnant pigs. The results suggest that transport of glycine into the uterine lumen and across the chorioallantois by SLC6A9 may be critical for delivery of this amino acid that is essential for conceptus development.

Despite the common usage of monosodium glutamate (MSG) as flavor enhancer, its excessive consumption has been linked to oxidative stress, inflammatory responses, and disruptions in cellular function, including within the reproductive systems. This study aimed to evaluate the protective role of Vitamin E against monosodium glutamate-induced utero-toxicity in Wistar rats. Twenty-four adult Wistar rats were randomly divided into four groups: Group A served as Control and received 1 ml of distilled water; Group B was administered 200 mg/kg of monosodium glutamate only; Group C received 100 IU/kg body weight of vitamin E and 200 mg/kg of monosodium glutamate; and Group D was given 100 IU/kg body weight of vitamin E only. Administration lasted for 28 days, after which the animals were sacrificed and the uterus were collected for histological assessments. Results from this study showed a significant increase (P<0.05) in final body weight when compared to the initial body weight in the Control and Vitamin E only groups. There was a significant decrease (P<0.05) in body weight changes in the MSG+ Vit. E and the Vitamin E only groups when compared to control. There was no significant difference (P>0.05) in uterine weight and utero-somatic index across the experimental groups. Histological assessment showed that MSG induced histological alterations evidenced as dilated uterine lumen, thin endometrium, and myometrial layers as well as reduced number and size of endometrial. However, in the group given MSG + vitamin E, there was significant improvement as evidenced by normal histological features similar to the control group; slit-like uterine lumen, endometrium containing several endometrial glands in the lamina propria, myometrium and perimetrium. In conclusion, findings from this study shows that Vitamin E attenuates MSG-induced uterotoxicity in Wistar rats.

The biology and morphology of the uterus were explored at approximately one month postpartum for cows that were diagnosed with metritis (uterine disease; n = 18) or healthy (n = 17) at 7-10 days postpartum and either treated or untreated with ceftiofur hydrochloride (2 × 2 factorial). Cows were slaughtered at one month postpartum, and the uterine horns were flushed to identify cows with a purulent or clear (non-purulent) content. RNA was isolated from caruncular and inter-caruncular endometria for RNA sequencing. Light microscopy was used to quantify inflammation, and scanning electron microscopy was used to assess the closure of the luminal surface of the uterus and the morphology of uterine glands. Results were that postpartum antibiotic treatment did not affect the endpoints that were measured including gene expression and morphology of the uterus. The primary biological driver of uterine function and morphology was inflammation at one month postpartum (endometritis; purulent material in the uterine lumen). Inflammation decreased epithelial growth factor signaling pathway expression (Wnt, Hippo, and Hedgehog) and biological processes involved in the formation of ciliated cells. Based on morphology, epithelial cells closed open areas of a denuded uterine surface using a process that involved depolarization and cell movement (planar cell polarity). Non-canonical Wnt signaling drives planar cell polarity, and the loss of Wnt signaling may explain the slower tissue repair in cows with endometritis. Future studies and therapeutic treatments should consider uterine involution as a process of wound repair and tissue regeneration in the face of bacterial infection.

Short- and long-term effects of uterine disease on oocyte developmental capacity in postpartum dairy cows.

The present investigation was carried out on clinical cases brought to the Teaching Veterinary Clinical Complex, College of veterinary Science and Animal husbandry, Mhow and in private clinics across Indore (M.P.). Eighteen female dogs (six in each group) of variable age brought with the history of vaginal discharge and or associated with clinical symptoms (depression, in-appetence and abdominal enlargement) with CEHP Complex were thoroughly investigated and on confirmation of the ailment, they were randomly allocated in the following 3 treatment groups viz. 1, 2 and 3. Each group comprises of 6 female dogs. The efficacy of treatment was judged on the basis of cessation of vaginal discharge, reduction in uterine lumen diameter and drift in haemato-biochemical parameters. Based on the results obtained in the present study it could be concluded that Haemato-biochemical parameters are of prognostic value. In this study group 1 (Cabergoline + Ciprofloxacin) was considered most effective with cessation of vaginal discharge by 8th day after treatment and significant difference in Haemato-biochemical parameters before and after treatment was observed

In briefDuring uterine receptivity, exocytosis from uterine epithelial cells (UECs) contributes to uterine fluid composition, playing a role in communication with an implanting blastocyst. SNAREs, a family of proteins involved in exocytosis, are increased in the receptive uterine epithelium of the rat.Uterine luminal fluid is composed of secretions from the uterine luminal and glandular epithelial cells. The fluid composition plays a role in cell-to-cell communication between the receptive endometrium and an invading blastocyst. Part of this fluid is released from the epithelial cells via exocytosis, mechanisms regulating this are not yet understood. Using transmission electron microscopy, this study identified extracellular vesicles in the uterine lumen at the time of fertilisation and uterine receptivity. Immunofluorescence microscopy showed SNARE proteins syntaxin 2 and SNAP23 in the apical area of UECs at the time of receptivity. SNAP23 was also found in the uterine fluid on day 5.5 of early pregnancy. Western blotting of isolated UECs demonstrated a significant increase in syntaxin 2 and SNAP23 at the time of uterine receptivity compared to the time of fertilisation. The morphological evidence of extracellular vesicles in the uterine lumen and the presence of SNARE proteins syntaxin 2 and SNAP23 in the apical part of the luminal epithelium at the time of uterine receptivity suggests that exocytosis contributes to the composition of the uterine luminal fluid, a potential component of maternal–foetal communication during early pregnancy.

Uterine diseases in cattle are frequently linked to bacterial infections, with pathogens commonly isolated from the uterine lumen. Bovine Gammaherpesvirus Type 4 (BoGHV-4) is notably prevalent in certain regions of Argentina and is associated with uterine diseases in postpartum cattle. This study aims to evaluate the impact of platelet-rich plasma (PRP) on the gene expression related to BoGHV-4 infection in the presence of lipopolysaccharide (LPS), exploring the potential of PRP as a therapeutic alternative. The interaction between LPS and Toll-like receptor 4 (TLR4) plays a crucial role in inflammatory responses, triggering cytokine production and immune activation. Our results show that PRP modulates TLR4 and TNF-α gene expression, indicating a potential inhibitory role in inflammatory processes. Furthermore, PRP alter the temporal dynamics of BoGHV-4 replication by modulating the expression of the viral immediate-early gene (IE-2) and delaying proinflammatory cytokine responses such as IL-8. Notably, PRP enhances IFN-γ expression, which could help prevent tissue damage caused by bacterial and viral coinfection. These findings highlight the potential of PRP as an anti-inflammatory agent with therapeutic benefits in treating uterine diseases, offering an alternative to traditional antibiotic treatments.

Background/Objectives: The aim of the present study was to explore the histopathological effects and tissue Vascular Endothelial Growth Factor (VEGF) levels of filgrastim and hyaluronic acid treatment in a rat model with experimentally induced Asherman syndrome. Methods: In this study, 26 female Sprague Dawley rats were used. First, a rat model of Asherman syndrome was established in two rats, and the remaining rats were randomly divided into three groups. A total of 0.1 mL trichloroacetic acid was applied to the right uterine horns of all groups to induce adhesion formation.Group I received no treatment, Group II received intrauterine hyaluronic acid treatment (0.01), Group III received subcutaneous Filgrastim treatment (50 μg/kg/day), and Group IV received both intrauterine hyaluronic acid and subcutaneous Filgrastim treatment. Histopathological analysis of uterine horns in the rats with and without Asherman syndrome, inflammation, glandular count, and fibrosis levels were examined. Tissue VEGF levels were investigated immunohistochemically. Results: Hyaluronic acid treatment resulted in an increase only in uterine lumen diameter and VEGF levels, while Filgrastim treatment led to an increase in uterine wall diameter, lumen diameter, gland count, and VEGF levels, as well as a decrease in fibrosis and inflammation scores. Combined treatment with filgrastim and hyaluronic acid showed an increase in lumen diameter, gland count, and VEGF levels, along with a decrease in inflammation and fibrosis scores (p < 0.05). Filgrastim treatment resulted in better effects for Asherman syndrome compared to hyaluronic acid treatment. There were no beneficial effects seen with the combined therapy. Conclusions: Filgrastim treatment resulted in better outcomes for Asherman syndrome compared to hyaluronic acid treatment. The combined therapy did not show additional benefits beyond what was achieved with Filgrastim treatment alone.

Pregnancy presents specific metabolic demands, and disruption caused by a high-fat high-sugar diet (HFHSD) have been associated with significant complications, including maternal health risk, fetal developmental issues, and infertility. Obesity-related changes in the uterine tissues may contribute to these challenges. This study analyzed structural changes in the uterus and adipose tissue of pregnant rats on gestation day 22 fed an HFHSD using various staining techniques. Hematoxylin and eosin staining showed morphological changes in the adipose tissue and the uterine structure, including the lumen size and the thickness of the myometrium, endometrium, and perimetrium. The amount of collagen in the uterus was determined by PicroSirius red staining, while PAS-D staining was used to observe glycogen content. Key protein expressions, such as insulin and leptin receptors and UCP1 and UCP3, were analyzed by immunohistochemistry. The HFHSD promoted hypertrophy of visceral and gonadal adipocytes, suggesting metabolic alterations. By the end of pregnancy, a significant reduction in uterine lumen size was observed. Additionally, a decrease in insulin and higher leptin receptor expressions in the myometrium indicated significant physiological alteration. These findings offer insight into how an HFHSD affects uterine structure and function during late pregnancy but should be interpreted within the physiological context of gestation-related metabolic changes. Further research is needed to understand the functional consequences of these alterations on reproductive and metabolic health.