Pub Date : 2025-11-01DOI: 10.1016/j.xfss.2025.07.004
Carolyn A. Nietupski Ph.D. , Megan R. Sax M.D. , Rose Dean B.S., M.B.A. , Andreja Moset Zupan B.S. , Emily G. Hurley M.D. , Stacey C. Schutte Ph.D.
Objective
To study how compressive forces influence fibroid and myometrial cells. Our work aimed to identify proteins and signaling pathways that are altered in fibroids in response to compressive forces.
Design
Laboratory-based.
Subjects
Patient-matched fibroid and myometrial cells were isolated from five women undergoing hysterectomy or myomectomy for the treatment of uterine fibroids. Only samples from women who had not had hormonal modulation within 3 months of surgery were used for this study. An embedded spheroid model was developed to model the fibroid tissue and provide a cushion that would help with the distribution of compressive force.
Exposure
Weights, 0 or 6.4 mm Hg, were added on top of an agarose cushion. Spheroids were cultured for 7 days.
Main Outcome Measures
Histological evaluation, RNA-sequencing (n = 5), and proteomics characterization (n = 3). Paired multi-test t-tests were performed for statistical analysis. Differentially expressed genes (DEGs) were considered clinically relevant if the same genes were also significantly differentially expressed in at least one of the four existing fibroid and myometrium RNA-sequencing datasets.
Results
A total of 61 clinically relevant DEGs were identified between cell types that were only differentially expressed when the spheroids were under compression. This included EPHB1 which encodes ephrin signaling receptor EphB1; it was upregulated log2 fold-change of 2.81 in fibroid cells (q = 5.35 × 10-3). Compression led to the enrichment of genes involved in extracellular matrix (ECM) organization; however, the genes varied between the cell types. At the protein level, myometrial spheroids had alterations in proteins associated with uterine fibroids (q = 1.00 × 10–33). There were alterations in collagen abundance in fibroid spheroids, but not collagen 1, although the collagenase MMP-1 was significantly lower in fibroid spheroids. Enrichment analysis identified ECM-receptor interactions as enriched in compression-induced changes between the cell types.
Conclusions
Compressive forces must be considered to study some of the important differences between fibroids and myometrium, including ephrin signaling. Enrichment analysis of the proteins with different abundances suggests that compression may also be involved in fibroid tumor initiation.
{"title":"Compressive force induces differential gene and protein expression in uterine fibroids","authors":"Carolyn A. Nietupski Ph.D. , Megan R. Sax M.D. , Rose Dean B.S., M.B.A. , Andreja Moset Zupan B.S. , Emily G. Hurley M.D. , Stacey C. Schutte Ph.D.","doi":"10.1016/j.xfss.2025.07.004","DOIUrl":"10.1016/j.xfss.2025.07.004","url":null,"abstract":"<div><h3>Objective</h3><div>To study how compressive forces influence fibroid and myometrial cells. Our work aimed to identify proteins and signaling pathways that are altered in fibroids in response to compressive forces.</div></div><div><h3>Design</h3><div>Laboratory-based.</div></div><div><h3>Subjects</h3><div>Patient-matched fibroid and myometrial cells were isolated from five women undergoing hysterectomy or myomectomy for the treatment of uterine fibroids. Only samples from women who had not had hormonal modulation within 3 months of surgery were used for this study. An embedded spheroid model was developed to model the fibroid tissue and provide a cushion that would help with the distribution of compressive force.</div></div><div><h3>Exposure</h3><div>Weights, 0 or 6.4 mm Hg, were added on top of an agarose cushion. Spheroids were cultured for 7 days.</div></div><div><h3>Main Outcome Measures</h3><div>Histological evaluation, RNA-sequencing (n = 5), and proteomics characterization (n = 3). Paired multi-test t-tests were performed for statistical analysis. Differentially expressed genes (DEGs) were considered clinically relevant if the same genes were also significantly differentially expressed in at least one of the four existing fibroid and myometrium RNA-sequencing datasets.</div></div><div><h3>Results</h3><div>A total of 61 clinically relevant DEGs were identified between cell types that were only differentially expressed when the spheroids were under compression. This included EPHB1 which encodes ephrin signaling receptor EphB1; it was upregulated log2 fold-change of 2.81 in fibroid cells (<em>q</em> = 5.35 × 10<sup>-3</sup>). Compression led to the enrichment of genes involved in extracellular matrix (ECM) organization; however, the genes varied between the cell types. At the protein level, myometrial spheroids had alterations in proteins associated with uterine fibroids (<em>q</em> = 1.00 × 10<sup>–33</sup>). There were alterations in collagen abundance in fibroid spheroids, but not collagen 1, although the collagenase MMP-1 was significantly lower in fibroid spheroids. Enrichment analysis identified ECM-receptor interactions as enriched in compression-induced changes between the cell types.</div></div><div><h3>Conclusions</h3><div>Compressive forces must be considered to study some of the important differences between fibroids and myometrium, including ephrin signaling. Enrichment analysis of the proteins with different abundances suggests that compression may also be involved in fibroid tumor initiation.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 463-474"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144710108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<div><h3>Objective</h3><div>To study the effect of laparoscopic ovarian drilling (LOD) vs. step-up gonadotropin therapy on follicular response in infertile anovulatory polycystic ovary syndrome (PCOS) women resistant to sequential letrozole + human menopausal gonadotropin (HMG)–based ovulation induction (OVI) cycles.</div></div><div><h3>Design</h3><div>Open-labeled, pilot, randomized controlled trial.</div></div><div><h3>Subjects</h3><div>Infertile anovulatory PCOS women (diagnosed according to modified Rotterdam criteria), resistant to sequential letrozole 5 mg + HMG-based OVI cycle (no dominant follicle >10 mm after 14 days of stimulation). Other inclusion criteria were: age 19–38 years; body mass index ≤35 kg/m<sup>2</sup>; patent fallopian tubes documented on either hysterosalpingography/saline infusion sonography or laparoscopy; antimüllerian hormone (AMH) levels >6 ng/mL. Exclusion criteria were AMH ≤6 ng/mL; moderate to severe male factor infertility and endometriosis.</div></div><div><h3>Intervention</h3><div>Participants in group 1 (N = 35) underwent LOD, after which OVI cycles were started with letrozole 5 mg from the following menses. Gonadotropin were added in a sequential manner if required as per the follicular response. Women in group 2 (N = 35) were administered injection of HMG (75 IU) from day 2 of menses and dose increments were done from day 9 onward as per response.</div></div><div><h3>Main Outcome Measures</h3><div>Primary outcome was follicular response (dominant follicle >16 mm). Secondary objectives were gonadotropin requirement per cycle, duration of stimulation, time to conception (months), clinical pregnancy rate and ongoing pregnancy rate (>12 weeks). The study also compared the effect of LOD on hormonal parameters (AMH, serum testosterone) and metabolic parameters (fasting insulin, fasting blood glucose, lipid profile, homeostasis model assessment of insulin resistance) after 1–2 months of procedure.</div></div><div><h3>Results</h3><div>Majority of the study participants (82.85%) belonged to PCOS phenotype A. The baseline clinical, hormonal, and metabolic characteristics and phenotype distribution were comparable in both groups. The follicular response was significantly higher in the LOD group (93.25%; 83/89) compared with step-up gonadotropin group (28.20%; 11/39). With four spontaneous conceptions, the median time to conception in LOD group was 3.9 (0–8.4) months. The clinical pregnancy rate per patient was significantly higher in LOD group [54.28% (19/35)] as compared with step-up gonadotropin group [8.57% (3/35)]. The ongoing pregnancy rate in the LOD group was 45.71% (16/35) vs. 0% (0/35) in the gonadotropin group. There was a significant fall in the AMH levels from 15.2 ± 2.7 ng/mL to 10.2 ± 4.4 ng/mL after LOD. Although statistically insignificant, the levels of luteinizing hormone/follicle-stimulating hormone ratio, testosterone, fasting insulin, fasting glucose and homeostasis model assessment of
{"title":"Laparoscopic ovarian drilling vs. step-up gonadotropin therapy in infertile anovulatory polycystic ovary syndrome women resistant to sequential letrozole and gonadotropin-based ovulation induction cycles: a randomized controlled trial","authors":"H.V. Bhavana M.D. , Reeta Mahey M.D., D.N.B. , Aarthi K. Jayraj M.D., D.N.B. , Ashish Datt Upadhyay Ph.D. , Archana Kumari M.D. , Garima Kachhawa M.D. , Ayushi Negi M.D. , Khushbu Bashir M.D. , Srikar Yedlapalli M.S.","doi":"10.1016/j.xfss.2025.07.006","DOIUrl":"10.1016/j.xfss.2025.07.006","url":null,"abstract":"<div><h3>Objective</h3><div>To study the effect of laparoscopic ovarian drilling (LOD) vs. step-up gonadotropin therapy on follicular response in infertile anovulatory polycystic ovary syndrome (PCOS) women resistant to sequential letrozole + human menopausal gonadotropin (HMG)–based ovulation induction (OVI) cycles.</div></div><div><h3>Design</h3><div>Open-labeled, pilot, randomized controlled trial.</div></div><div><h3>Subjects</h3><div>Infertile anovulatory PCOS women (diagnosed according to modified Rotterdam criteria), resistant to sequential letrozole 5 mg + HMG-based OVI cycle (no dominant follicle >10 mm after 14 days of stimulation). Other inclusion criteria were: age 19–38 years; body mass index ≤35 kg/m<sup>2</sup>; patent fallopian tubes documented on either hysterosalpingography/saline infusion sonography or laparoscopy; antimüllerian hormone (AMH) levels >6 ng/mL. Exclusion criteria were AMH ≤6 ng/mL; moderate to severe male factor infertility and endometriosis.</div></div><div><h3>Intervention</h3><div>Participants in group 1 (N = 35) underwent LOD, after which OVI cycles were started with letrozole 5 mg from the following menses. Gonadotropin were added in a sequential manner if required as per the follicular response. Women in group 2 (N = 35) were administered injection of HMG (75 IU) from day 2 of menses and dose increments were done from day 9 onward as per response.</div></div><div><h3>Main Outcome Measures</h3><div>Primary outcome was follicular response (dominant follicle >16 mm). Secondary objectives were gonadotropin requirement per cycle, duration of stimulation, time to conception (months), clinical pregnancy rate and ongoing pregnancy rate (>12 weeks). The study also compared the effect of LOD on hormonal parameters (AMH, serum testosterone) and metabolic parameters (fasting insulin, fasting blood glucose, lipid profile, homeostasis model assessment of insulin resistance) after 1–2 months of procedure.</div></div><div><h3>Results</h3><div>Majority of the study participants (82.85%) belonged to PCOS phenotype A. The baseline clinical, hormonal, and metabolic characteristics and phenotype distribution were comparable in both groups. The follicular response was significantly higher in the LOD group (93.25%; 83/89) compared with step-up gonadotropin group (28.20%; 11/39). With four spontaneous conceptions, the median time to conception in LOD group was 3.9 (0–8.4) months. The clinical pregnancy rate per patient was significantly higher in LOD group [54.28% (19/35)] as compared with step-up gonadotropin group [8.57% (3/35)]. The ongoing pregnancy rate in the LOD group was 45.71% (16/35) vs. 0% (0/35) in the gonadotropin group. There was a significant fall in the AMH levels from 15.2 ± 2.7 ng/mL to 10.2 ± 4.4 ng/mL after LOD. Although statistically insignificant, the levels of luteinizing hormone/follicle-stimulating hormone ratio, testosterone, fasting insulin, fasting glucose and homeostasis model assessment of","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 453-462"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144786088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-01DOI: 10.1016/j.xfss.2025.07.003
Edson Borges Jr. Ph.D. , Daniela Braga Ph.D. , Maite del Collado Ph.D. , Assumpto Iaconelli Jr. M.D. , Jullin Fjeldstad M.Sc. , Natalie Mercuri M.Sc. , Parisa Mojiri M.Sc. , Amanda Setti M.Sc.
Objective
To study whether artificial intelligence (AI)–driven oocyte evaluation is associated with blastocyst development and quality in couples with severe male factor infertility (SMF) undergoing intracytoplasmic sperm injection (ICSI) cycles.
Design
Cohort study.
Subjects
Fourteen thousand six hundred two oocyte images from 2,156 ICSI cycles performed between January 2020 and May 2024 in a private, university-affiliated in vitro fertilization center. Cycles were categorized into the following two groups: SMF (n = 200 cycles, 1,478 embryos) and non-SMF (n = 1,956 cycles, 13,124 embryos). Severe male factor infertility was defined as <5 million sperm in the ejaculate.
Exposure
Oocyte images were captured before ICSI and scored using the AI tool MAGENTA. The predictive value of Magenta Scores (MS) on embryonic development was assessed. The association between MS and oocyte fertilization and blastocyst formation was analyzed.
Main Outcome Measures
Oocyte fertilization, blastulation rate, and blastocyst quality.
Results
Magenta scores were significantly lower in oocytes that failed to fertilize compared with those that successfully fertilized (5.00 ± 0.04 vs. 6.44 ± 0.03). Blastulation rate was lower in the SMF group (46.61% vs. 50.80%), and blastocysts exhibited higher MS than nonblastocysts (5.12 ± 0.3 vs. 6.69 ± 0.3). The top-quality blastocyst rate was lower in SMF (56.6% vs. 65.2%), and high-quality blastocysts had higher MS than lower-quality ones (7.2 ± 0.6 vs. 6.8 ± 0.5). Among SMF cycles, MS were lower in oocytes that failed to fertilize (4.91 ± 0.12 vs. 6.34 ± 0.10). Magenta scores also differed between embryos that reached the blastocyst stage and those that did not (6.70 ± 0.11 vs. 4.96 ± 0.10). Top-quality blastocysts had significantly higher MS than others (7.00 ± 0.21 vs. 6.39 ± 0.19). Paternal age negatively correlated with fertilization, blastulation, and blastocyst quality; however, differences remained significant after adjusting for paternal age.
Conclusion
Artificial intelligence–based oocyte evaluation is associated with fertilization, blastulation, and blastocyst quality in SMF couples undergoing ICSI cycles. Magenta score values were consistently higher for blastocysts than nonblastocysts, demonstrating the AI tool’s utility in identifying oocytes with greater developmental potential, regardless of male infertility factors. However, the absence of sperm-specific factors in the MAGENTA algorithm may limit its ability to fully account for male infertility.
{"title":"Artificial intelligence–driven oocyte assessment for predicting blastulation and high-quality blastocyst formation in severe male factor infertility","authors":"Edson Borges Jr. Ph.D. , Daniela Braga Ph.D. , Maite del Collado Ph.D. , Assumpto Iaconelli Jr. M.D. , Jullin Fjeldstad M.Sc. , Natalie Mercuri M.Sc. , Parisa Mojiri M.Sc. , Amanda Setti M.Sc.","doi":"10.1016/j.xfss.2025.07.003","DOIUrl":"10.1016/j.xfss.2025.07.003","url":null,"abstract":"<div><h3>Objective</h3><div>To study whether artificial intelligence (AI)–driven oocyte evaluation is associated with blastocyst development and quality in couples with severe male factor infertility (SMF) undergoing intracytoplasmic sperm injection (ICSI) cycles.</div></div><div><h3>Design</h3><div>Cohort study.</div></div><div><h3>Subjects</h3><div>Fourteen thousand six hundred two oocyte images from 2,156 ICSI cycles performed between January 2020 and May 2024 in a private, university-affiliated in vitro fertilization center. Cycles were categorized into the following two groups: SMF (n = 200 cycles, 1,478 embryos) and non-SMF (n = 1,956 cycles, 13,124 embryos). Severe male factor infertility was defined as <5 million sperm in the ejaculate.</div></div><div><h3>Exposure</h3><div>Oocyte images were captured before ICSI and scored using the AI tool MAGENTA. The predictive value of Magenta Scores (MS) on embryonic development was assessed. The association between MS and oocyte fertilization and blastocyst formation was analyzed.</div></div><div><h3>Main Outcome Measures</h3><div>Oocyte fertilization, blastulation rate, and blastocyst quality.</div></div><div><h3>Results</h3><div>Magenta scores were significantly lower in oocytes that failed to fertilize compared with those that successfully fertilized (5.00 ± 0.04 vs. 6.44 ± 0.03). Blastulation rate was lower in the SMF group (46.61% vs. 50.80%), and blastocysts exhibited higher MS than nonblastocysts (5.12 ± 0.3 vs. 6.69 ± 0.3). The top-quality blastocyst rate was lower in SMF (56.6% vs. 65.2%), and high-quality blastocysts had higher MS than lower-quality ones (7.2 ± 0.6 vs. 6.8 ± 0.5). Among SMF cycles, MS were lower in oocytes that failed to fertilize (4.91 ± 0.12 vs. 6.34 ± 0.10). Magenta scores also differed between embryos that reached the blastocyst stage and those that did not (6.70 ± 0.11 vs. 4.96 ± 0.10). Top-quality blastocysts had significantly higher MS than others (7.00 ± 0.21 vs. 6.39 ± 0.19). Paternal age negatively correlated with fertilization, blastulation, and blastocyst quality; however, differences remained significant after adjusting for paternal age.</div></div><div><h3>Conclusion</h3><div>Artificial intelligence–based oocyte evaluation is associated with fertilization, blastulation, and blastocyst quality in SMF couples undergoing ICSI cycles. Magenta score values were consistently higher for blastocysts than nonblastocysts, demonstrating the AI tool’s utility in identifying oocytes with greater developmental potential, regardless of male infertility factors. However, the absence of sperm-specific factors in the MAGENTA algorithm may limit its ability to fully account for male infertility.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 426-432"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144661170","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-01DOI: 10.1016/j.xfss.2025.08.003
Valerie A. Flores M.D., Pablo A. Delis M.D., Ramanaiah Mamillapalli Ph.D., Hugh S. Taylor M.D.
Objective
Asherman syndrome (AS) is characterized by intrauterine adhesions/fibrosis, resulting from damage to the endometrial basalis layer. The consequences of AS include infertility, recurrent pregnancy loss, preterm rupture of membranes, and placental abruption. Hysteroscopic adhesiolysis does not consistently restore endometrial function; there is a need for more effective treatments. Bone marrow–derived mesenchymal stem cells (BMD-MSCs) circulate systemically and contribute to tissue repair/regeneration, suggesting that they may serve as a source of progenitor cells for endometrial regeneration. Increasing the supply of BMD-MSCs to the endometrium may treat AS by allowing for regeneration/replenishment of endometrial progenitor cells. Mobilization of autologous BMD-MSCs using AMD3100, a CXC-motif-receptor-4 antagonist, is approved for bone marrow transplantation. We aimed to determine whether the optimal timing of AMD3100 administration—on the basis of CXCL12 production—would better recruit BMD-MSCs in a murine model of severe AS and restore functioning endometrium/fertility.
Design
Severe AS murine model.
Subjects
C57BL/6 mice in the diestrus phase undergoing surgical AS induction.
Exposure
Single injection with AMD3100 (treatment) or vehicle (saline). AMD3100 administration timing was based on the determination of maximum CXCL12 release after AS induction. Mice were then mated.
Main Outcome Measures
Time to pregnancy, litter size, and miscarriage rate.
Results
Maximum uterine CXCL12 production occurred 48 hours after AS induction; thus, AMD3100 vs. saline was administered 48 hours after induction. Of the AMD3100-treated mice, all achieved pregnancy and delivered. The treatment group became pregnant and delivered significantly sooner, indicating a faster time to conception (20 vs. 26 days). The treatment group had significantly larger litter sizes (6.5 vs. 4.2 pups) and significantly more live pups at delivery than the control group (6.0 vs. 2.7).
Conclusion
AMD3100 had a significant effect on uterine repair and regeneration in AS. The likelihood of pregnancy was significantly higher and more rapid in AS mice treated with AMD3100. Treated mice also had larger litter sizes and fewer miscarriages than controls. Furthermore, we determined for the first time the levels of CXCL12 in uteri after uterine injury, which allowed for the determination of the optimal timing of AMD3100 administration, to ensure mobilized BMD-MSCs homed to the uterus.
目的：Asherman综合征（AS）以子宫内膜基底层损伤引起的宫内粘连/纤维化为特征。AS的后果包括不孕症、反复流产、胎膜早破和胎盘早剥。宫腔镜下粘连松解术不能始终恢复子宫内膜功能；需要更有效的治疗方法。骨髓间充质干细胞（BM-MSCs）具有系统性循环，有助于组织修复/再生，这表明它们可能是子宫内膜再生的祖细胞来源。增加向子宫内膜提供BM-MSCs可能通过允许子宫内膜祖细胞的再生/补充来治疗AS。使用AMD3100（一种CXC-motif-receptor-4拮抗剂）动员自体骨髓间充质干细胞被批准用于骨髓移植。我们的目的是确定基于CXCL12生成的AMD 3100给药的最佳时间是否能更好地在严重AS小鼠模型中招募BM-MSCs，并恢复功能子宫内膜/生育能力。设计：重度AS小鼠模型研究对象：处于绝育期接受手术诱导的C57BL/6小鼠。暴露：单次注射AMD3100（治疗）或载体（生理盐水）。AMD3100给药时间基于AS诱导后CXCL12最大释放量的测定。然后对老鼠进行交配。测量的主要结果：妊娠时间、产仔数和流产率。结果：AS诱导后48小时子宫CXCL12产生量最大，因此在诱导后48小时给予AMD3100与生理盐水对照。在接受AMD3100治疗的小鼠中，所有小鼠都成功怀孕并分娩。治疗组怀孕和分娩明显更快，表明受孕时间更快（20天对26天）。治疗组产仔数明显大于对照组（6.5 vs 4.2），分娩时活仔数明显多于对照组（6.0 vs 2.7）。结论：AMD3100对AS患者的子宫修复和再生有明显的促进作用。用AMD3100治疗的AS小鼠，怀孕的可能性明显更高、更快。与对照组相比，接受治疗的小鼠产仔量更大，流产率更低。此外，我们首次测定了子宫损伤后子宫内CXCL12的水平，从而确定了AMD3100给药的最佳时机，以确保动员的BMD-MSCs回到子宫。
{"title":"Use of AMD3100 for bone marrow–derived mesenchymal stem cell mobilization in the treatment of murine Asherman's syndrome","authors":"Valerie A. Flores M.D., Pablo A. Delis M.D., Ramanaiah Mamillapalli Ph.D., Hugh S. Taylor M.D.","doi":"10.1016/j.xfss.2025.08.003","DOIUrl":"10.1016/j.xfss.2025.08.003","url":null,"abstract":"<div><h3>Objective</h3><div>Asherman syndrome (AS) is characterized by intrauterine adhesions/fibrosis, resulting from damage to the endometrial basalis layer. The consequences of AS include infertility, recurrent pregnancy loss, preterm rupture of membranes, and placental abruption. Hysteroscopic adhesiolysis does not consistently restore endometrial function; there is a need for more effective treatments. Bone marrow–derived mesenchymal stem cells (BMD-MSCs) circulate systemically and contribute to tissue repair/regeneration, suggesting that they may serve as a source of progenitor cells for endometrial regeneration. Increasing the supply of BMD-MSCs to the endometrium may treat AS by allowing for regeneration/replenishment of endometrial progenitor cells. Mobilization of autologous BMD-MSCs using AMD3100, a CXC-motif-receptor-4 antagonist, is approved for bone marrow transplantation. We aimed to determine whether the optimal timing of AMD3100 administration—on the basis of CXCL12 production—would better recruit BMD-MSCs in a murine model of severe AS and restore functioning endometrium/fertility.</div></div><div><h3>Design</h3><div>Severe AS murine model.</div></div><div><h3>Subjects</h3><div>C57BL/6 mice in the diestrus phase undergoing surgical AS induction.</div></div><div><h3>Exposure</h3><div>Single injection with AMD3100 (treatment) or vehicle (saline). AMD3100 administration timing was based on the determination of maximum CXCL12 release after AS induction. Mice were then mated.</div></div><div><h3>Main Outcome Measures</h3><div>Time to pregnancy, litter size, and miscarriage rate.</div></div><div><h3>Results</h3><div>Maximum uterine CXCL12 production occurred 48 hours after AS induction; thus, AMD3100 vs. saline was administered 48 hours after induction. Of the AMD3100-treated mice, all achieved pregnancy and delivered. The treatment group became pregnant and delivered significantly sooner, indicating a faster time to conception (20 vs. 26 days). The treatment group had significantly larger litter sizes (6.5 vs. 4.2 pups) and significantly more live pups at delivery than the control group (6.0 vs. 2.7).</div></div><div><h3>Conclusion</h3><div>AMD3100 had a significant effect on uterine repair and regeneration in AS. The likelihood of pregnancy was significantly higher and more rapid in AS mice treated with AMD3100. Treated mice also had larger litter sizes and fewer miscarriages than controls. Furthermore, we determined for the first time the levels of CXCL12 in uteri after uterine injury, which allowed for the determination of the optimal timing of AMD3100 administration, to ensure mobilized BMD-MSCs homed to the uterus.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 475-482"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144862664","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To investigate the potential of recombinant interleukin-22 (rIL-22) in preventing inflammation-triggered abortion.
Design
Experimental animal study using wild type (WT, C57BL/6J) and IL-22 knockout (IL-22⁻/⁻) mice aged 7–14 weeks. Abortion was induced via intraperitoneal injection of lipopolysaccharide (LPS) on gestational day 8.5.
Subjects
Pregnant WT and IL-22⁻/⁻ mice (n = 6/group for tissue collection at gestational day 8.5+48 hours; n = 3/group for monitoring to delivery at gestational day 20.5).
Intervention(s)
Tail vein injection of recombinant IL-22 at varying doses (5, 10, or 20 µg/mouse) prior to LPS administration.
Main Outcome Measure(s)
Pregnancy outcomes (abortion vs. live delivery), endometrial mucosal architecture, epithelial barrier integrity (transepithelial electric resistance), gene expression profiles (qRT-PCR), protein distribution (IHC/IF), and immune marker levels in amniotic fluid (multiplex immunoassay).
Results
Administration of rIL-22 to IL-22⁻/⁻ mice restored endometrial mucosal architecture to levels similar to WT animals and prevented abortion in a dose-dependent manner. rIL-22 treatment rescued pregnancy in IL-22⁻/⁻ mice at 5–20 µg/mouse, whereas only ≤10 µg/mouse was effective in WT mice. High-dose rIL-22 (20 µg/mouse), when combined with LPS, negatively affected pregnancy outcomes in WT mice, and 20 µg alone resulted in adverse outcomes in both genotypes.
Conclusion
Interleukin-22 emerges as a promising therapeutic target in reproductive medicine, offering potential avenues for preserving pregnancy in the face of inflammatory challenges. However, increased levels of IL-22 could be associated with reproductive failures, emphasizing the delicate balance required for immune regulation. Our study underscores the importance of immune regulation in reproductive health and highlights IL-22 as a novel candidate for improving pregnancy outcomes in inflammatory conditions.
{"title":"The dual role of interleukin-22: balancing protection and risk in pregnancy during inflammatory challenges","authors":"Umida Ganieva Ph.D. , Mahmood Bilal Ph.D. , Ana Adam , Cecilia Pena-Rasgado Ph.D. , Wren Michaels Ph.D. , Hector Rasgado-Flores Ph.D. , Amy Thees Ph.D. , Thanh Luu M.D. , Joanne Kwak-Kim M.D. , Svetlana Dambaeva M.D., Ph.D.","doi":"10.1016/j.xfss.2025.09.001","DOIUrl":"10.1016/j.xfss.2025.09.001","url":null,"abstract":"<div><h3>Objective</h3><div>To investigate the potential of recombinant interleukin-22 (rIL-22) in preventing inflammation-triggered abortion.</div></div><div><h3>Design</h3><div>Experimental animal study using wild type (WT, C57BL/6J) and IL-22 knockout (IL-22⁻/⁻) mice aged 7–14 weeks. Abortion was induced via intraperitoneal injection of lipopolysaccharide (LPS) on gestational day 8.5.</div></div><div><h3>Subjects</h3><div>Pregnant WT and IL-22⁻/⁻ mice (n = 6/group for tissue collection at gestational day 8.5+48 hours; n = 3/group for monitoring to delivery at gestational day 20.5).</div></div><div><h3>Intervention(s)</h3><div>Tail vein injection of recombinant IL-22 at varying doses (5, 10, or 20 µg/mouse) prior to LPS administration.</div></div><div><h3>Main Outcome Measure(s)</h3><div>Pregnancy outcomes (abortion vs. live delivery), endometrial mucosal architecture, epithelial barrier integrity (transepithelial electric resistance), gene expression profiles (qRT-PCR), protein distribution (IHC/IF), and immune marker levels in amniotic fluid (multiplex immunoassay).</div></div><div><h3>Results</h3><div>Administration of rIL-22 to IL-22⁻/⁻ mice restored endometrial mucosal architecture to levels similar to WT animals and prevented abortion in a dose-dependent manner. rIL-22 treatment rescued pregnancy in IL-22⁻/⁻ mice at 5–20 µg/mouse, whereas only ≤10 µg/mouse was effective in WT mice. High-dose rIL-22 (20 µg/mouse), when combined with LPS, negatively affected pregnancy outcomes in WT mice, and 20 µg alone resulted in adverse outcomes in both genotypes.</div></div><div><h3>Conclusion</h3><div>Interleukin-22 emerges as a promising therapeutic target in reproductive medicine, offering potential avenues for preserving pregnancy in the face of inflammatory challenges. However, increased levels of IL-22 could be associated with reproductive failures, emphasizing the delicate balance required for immune regulation. Our study underscores the importance of immune regulation in reproductive health and highlights IL-22 as a novel candidate for improving pregnancy outcomes in inflammatory conditions.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 433-446"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145042420","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-01DOI: 10.1016/j.xfss.2025.08.004
Skylar G. Montague Redecke B.Sc. , Austin Bell-Hensley Ph.D. , MyeongJin Yi Ph.D. , Shuyun Li Ph.D. , Francesco J. DeMayo Ph.D.
Objective
To investigate enhancer-mediated regulation of progesterone receptor (PGR) isoforms, PGR-A and PGR-B, in human endometrial stromal cells, and to determine how isoform modulation shapes the progesterone-responsive transcriptome and cistrome relevant to endometrial function.
Design
A clustered regularly interspaced short palindromic repeats–based functional genomic screen was used to identify distal enhancers in telomerase-immortalized human endometrial stromal cells. Subsequent clustered regularly interspaced short palindromic repeats targeting of identified enhancers and the PGR promoter was used to modulate PGR isoform balance and assess functional consequences.
Subjects
None.
Exposure
Engineered endometrial stromal cells were treated with medroxyprogesterone acetate or vehicle.
Main Outcome Measures
PGR isoform expression was assessed by western blot, the progesterone-responsive transcriptome was characterized by bulk ribonucleic acid sequencing, and the PGR cistrome was characterized by Cut&Run.
Results
Two distal PGR enhancers were identified in endometrial stromal cells located approximately 60 and 220 kb upstream of the PGR transcription start site. Clustered regularly interspaced short palindromic repeats–based activation of these enhancers upregulated both PGR-A and PGR-B, whereas promoter activation primarily upregulated PGR-B. Bulk ribonucleic acid sequencing revealed that shifting the PGR isoform balance altered the progesterone-regulated transcriptome: PGR-A/B-equivalent cells exhibited proinflammatory gene signatures, whereas PGR-B-dominant cells demonstrated suppression of inflammatory signaling and altered cell cycle programs. The PGR Cut&Run profiling revealed distinct genomic binding patterns associated with each isoform profile. Integration of the PGR cistrome with chromatin interaction maps suggested that these isoforms directly regulate distinct gene subsets involved in inflammation and fibrosis. Mechanistically, estrogen receptor alpha (ESR1) indirectly activated PGR-A expression, potentially through recruitment of Forkhead box protein O1 (FOXO1) at the distal enhancer, suggesting a noncanonical, enhancer-mediated mechanism of PGR regulation.
Conclusions
Distal enhancers regulate the PGR isoform balance and shape the progesterone-responsive transcriptome in human endometrial stromal cells. This enhancer-mediated mechanism expands current models of PGR regulation beyond promoter-level control and may offer potential therapeutic targets to restore normal progesterone responsiveness in conditions marked by PGR isoform imbalance.
{"title":"Progesterone receptor isoform modulation via enhancer activation regulates progesterone signaling in endometrial stromal cells","authors":"Skylar G. Montague Redecke B.Sc. , Austin Bell-Hensley Ph.D. , MyeongJin Yi Ph.D. , Shuyun Li Ph.D. , Francesco J. DeMayo Ph.D.","doi":"10.1016/j.xfss.2025.08.004","DOIUrl":"10.1016/j.xfss.2025.08.004","url":null,"abstract":"<div><h3>Objective</h3><div>To investigate enhancer-mediated regulation of progesterone receptor (PGR) isoforms, PGR-A and PGR-B, in human endometrial stromal cells, and to determine how isoform modulation shapes the progesterone-responsive transcriptome and cistrome relevant to endometrial function.</div></div><div><h3>Design</h3><div>A clustered regularly interspaced short palindromic repeats–based functional genomic screen was used to identify distal enhancers in telomerase-immortalized human endometrial stromal cells. Subsequent clustered regularly interspaced short palindromic repeats targeting of identified enhancers and the PGR promoter was used to modulate PGR isoform balance and assess functional consequences.</div></div><div><h3>Subjects</h3><div>None.</div></div><div><h3>Exposure</h3><div>Engineered endometrial stromal cells were treated with medroxyprogesterone acetate or vehicle.</div></div><div><h3>Main Outcome Measures</h3><div>PGR isoform expression was assessed by western blot, the progesterone-responsive transcriptome was characterized by bulk ribonucleic acid sequencing, and the PGR cistrome was characterized by Cut&Run.</div></div><div><h3>Results</h3><div>Two distal PGR enhancers were identified in endometrial stromal cells located approximately 60 and 220 kb upstream of the PGR transcription start site. Clustered regularly interspaced short palindromic repeats–based activation of these enhancers upregulated both PGR-A and PGR-B, whereas promoter activation primarily upregulated PGR-B. Bulk ribonucleic acid sequencing revealed that shifting the PGR isoform balance altered the progesterone-regulated transcriptome: PGR-A/B-equivalent cells exhibited proinflammatory gene signatures, whereas PGR-B-dominant cells demonstrated suppression of inflammatory signaling and altered cell cycle programs. The PGR Cut&Run profiling revealed distinct genomic binding patterns associated with each isoform profile. Integration of the PGR cistrome with chromatin interaction maps suggested that these isoforms directly regulate distinct gene subsets involved in inflammation and fibrosis. Mechanistically, estrogen receptor alpha (ESR1) indirectly activated PGR-A expression, potentially through recruitment of Forkhead box protein O1 (FOXO1) at the distal enhancer, suggesting a noncanonical, enhancer-mediated mechanism of PGR regulation.</div></div><div><h3>Conclusions</h3><div>Distal enhancers regulate the PGR isoform balance and shape the progesterone-responsive transcriptome in human endometrial stromal cells. This enhancer-mediated mechanism expands current models of PGR regulation beyond promoter-level control and may offer potential therapeutic targets to restore normal progesterone responsiveness in conditions marked by PGR isoform imbalance.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 397-413"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144981031","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To compare the transcriptomes induced by estrogens used in combined oral contraceptives (COCs): estetrol (E4), ethinylestradiol (EE), and estradiol (E2) in human primary endometriotic stromal cells.
Design
Endometriotic stromal cells were cultured in the presence of E4 (10−6 mol/L), E2 (10−8 mol/L) or EE (10−9 mol/L) for 24 hours. Transcriptomes induced by E4 were compared with those induced by E2 and EE.
Subjects
Endometriotic stromal cells were obtained from patients with ovarian endometriomas.
Main Outcome Measures
Transcriptomes were examined using RNA sequencing analysis, and messenger RNA levels were measured using reverse transcription-quantitative polymerase chain reaction.
Results
Estetrol treatment resulted in 114 differentially expressed genes compared with the control (|log2fold change| >0.2). The number was higher than those differentially expressed between E4 and E2 (82 genes) and between E4 and EE (75 genes). Of the differentially expressed genes between E4 and the control, 79% (90 genes) changed in the same manner as in EE and E2. In contrast, 21% (24 genes) of the genes changed in an E4-preferential manner.
Conclusion
The present study elucidated that although E4 induces the expression of specific unique genes in endometriotic stromal cells distinct from those regulated by other estrogens, the overall gene expression profile is largely analogous to that induced by other estrogens. When differences in progestin are not considered, it is suggested that the COC containing E4 exhibits effects on endometriosis comparable with those of other COCs.
{"title":"Transcriptome landscapes induced by estetrol, estradiol, and ethinylestradiol in primary human endometriotic stromal cells","authors":"Marie Nakajima M.D. , Gentaro Izumi M.D., Ph.D. , Kaori Koga M.D., Ph.D. , Vita Silvana M.D. , Mohammed Elsherbini M.D., Ph.D. , Atsushi Okumura M.Sc. , Michihito Wada Ph.D. , Satoru Yamanaka Ph.D. , Yasushi Hirota M.D., Ph.D. , Yutaka Osuga M.D., Ph.D.","doi":"10.1016/j.xfss.2025.06.002","DOIUrl":"10.1016/j.xfss.2025.06.002","url":null,"abstract":"<div><h3>Objective</h3><div>To compare the transcriptomes induced by estrogens used in combined oral contraceptives (COCs): estetrol (E4), ethinylestradiol (EE), and estradiol (E2) in human primary endometriotic stromal cells.</div></div><div><h3>Design</h3><div>Endometriotic stromal cells were cultured in the presence of E4 (10<sup>−6</sup> mol/L), E2 (10<sup>−8</sup> mol/L) or EE (10<sup>−9</sup> mol/L) for 24 hours. Transcriptomes induced by E4 were compared with those induced by E2 and EE.</div></div><div><h3>Subjects</h3><div>Endometriotic stromal cells were obtained from patients with ovarian endometriomas.</div></div><div><h3>Main Outcome Measures</h3><div>Transcriptomes were examined using RNA sequencing analysis, and messenger RNA levels were measured using reverse transcription-quantitative polymerase chain reaction.</div></div><div><h3>Results</h3><div>Estetrol treatment resulted in 114 differentially expressed genes compared with the control (|log2fold change| >0.2). The number was higher than those differentially expressed between E4 and E2 (82 genes) and between E4 and EE (75 genes). Of the differentially expressed genes between E4 and the control, 79% (90 genes) changed in the same manner as in EE and E2. In contrast, 21% (24 genes) of the genes changed in an E4-preferential manner.</div></div><div><h3>Conclusion</h3><div>The present study elucidated that although E4 induces the expression of specific unique genes in endometriotic stromal cells distinct from those regulated by other estrogens, the overall gene expression profile is largely analogous to that induced by other estrogens. When differences in progestin are not considered, it is suggested that the COC containing E4 exhibits effects on endometriosis comparable with those of other COCs.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 387-396"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144512883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-01DOI: 10.1016/j.xfss.2025.05.002
Maria Luisa Pardiñas M.Sc. , Rocío Rivera-Egea Ph.D. , Jose Maria de los Santos Ph.D. , Carmen Vidal Ph.D. , Juan Giles Ph.D. , David Ortega-Jaen M.Sc. , Julia Gil M.Sc. , Angel Martin Ph.D. , Thamara Viloria Ph.D. , Maria Jose de los Santos Ph.D.
Objective
To determine the impact of a microfluidic-based sperm selection device on sperm parameters and embryo variables compared with the conventional swim-up method in sibling oocytes.
Design
Prospective observational study.
Subjects
A total of 345 oocytes were recruited from 27 couples undergoing intracytoplasmic sperm injection, including both own (n = 195) and donation (n = 150) cycles. None of the patients presented severe male factor.
Exposure
Semen sample was divided into 2 groups and processed using swim-up or the microfluidic sperm selection device (MSSD) ZyMōt. Half of the oocytes were inseminated with swim-up–selected spermatozoa, and the rest were inseminated with MSSD-selected spermatozoa. Embryo development was followed by time-lapse. Sperm deoxyribonucleic acid (DNA) fragmentation was measured using the sperm chromatin dispersion test, analyzed with ImageJ. A Mann-Whitney U test was performed for statistical analysis.
Main Outcome Measures
Sperm DNA fragmentation levels, sperm parameters, fertilization rates, embryo morphokinetics, and rate of usable blastocysts.
Results
Sperm DNA fragmentation was significantly lower in the MSSD group than in the swim-up group (20.3% vs. 11%), indicating a better selection. Analyzing separately the oocytes from the patient's own cycles, MSSD showed a significantly higher rate of usable blastocysts per fertilized oocyte. However, this difference was not observed using donated oocytes or when both cycles were combined. Embryos from the swim-up group showed a significant delay in time of pronuclear appearance and morula formation compared with those from the MSSD group, being more marked in donated oocytes. No significant differences were observed in the fertilization rate and the remaining morphokinetic times.
Conclusion
This study provides valuable information on the use of MSSD for noninvasive sperm selection. When MSSD was used, we observed a reduction in sperm DNA fragmentation and an enhancement in the number of usable embryos in our own cycles. These findings could be compatible with a reduced capacity to repair sperm damage due to poorer oocyte quality caused by advanced age.
{"title":"Impact of a microfluidic-based selection device on sperm deoxyribonucleic acid fragmentation and intracytoplasmic sperm injection cycles","authors":"Maria Luisa Pardiñas M.Sc. , Rocío Rivera-Egea Ph.D. , Jose Maria de los Santos Ph.D. , Carmen Vidal Ph.D. , Juan Giles Ph.D. , David Ortega-Jaen M.Sc. , Julia Gil M.Sc. , Angel Martin Ph.D. , Thamara Viloria Ph.D. , Maria Jose de los Santos Ph.D.","doi":"10.1016/j.xfss.2025.05.002","DOIUrl":"10.1016/j.xfss.2025.05.002","url":null,"abstract":"<div><h3>Objective</h3><div>To determine the impact of a microfluidic-based sperm selection device on sperm parameters and embryo variables compared with the conventional swim-up method in sibling oocytes.</div></div><div><h3>Design</h3><div>Prospective observational study.</div></div><div><h3>Subjects</h3><div>A total of 345 oocytes were recruited from 27 couples undergoing intracytoplasmic sperm injection, including both own (n = 195) and donation (n = 150) cycles. None of the patients presented severe male factor.</div></div><div><h3>Exposure</h3><div>Semen sample was divided into 2 groups and processed using swim-up or the microfluidic sperm selection device (MSSD) ZyMōt. Half of the oocytes were inseminated with swim-up–selected spermatozoa, and the rest were inseminated with MSSD-selected spermatozoa. Embryo development was followed by time-lapse. Sperm deoxyribonucleic acid (DNA) fragmentation was measured using the sperm chromatin dispersion test, analyzed with ImageJ. A Mann-Whitney <em>U</em> test was performed for statistical analysis.</div></div><div><h3>Main Outcome Measures</h3><div><span>Sperm DNA fragmentation levels, sperm parameters, fertilization rates, embryo morphokinetics, and rate of usable </span>blastocysts.</div></div><div><h3>Results</h3><div><span>Sperm DNA fragmentation was significantly lower in the MSSD group than in the swim-up group (20.3% vs. 11%), indicating a better selection. Analyzing separately the oocytes from the patient's own cycles, MSSD showed a significantly higher rate of usable blastocysts per fertilized oocyte. However, this difference was not observed using donated oocytes or when both cycles were combined. Embryos from the swim-up group showed a significant delay in time of pronuclear appearance and </span>morula formation compared with those from the MSSD group, being more marked in donated oocytes. No significant differences were observed in the fertilization rate and the remaining morphokinetic times.</div></div><div><h3>Conclusion</h3><div>This study provides valuable information on the use of MSSD for noninvasive sperm selection. When MSSD was used, we observed a reduction in sperm DNA fragmentation and an enhancement in the number of usable embryos in our own cycles. These findings could be compatible with a reduced capacity to repair sperm damage due to poorer oocyte quality caused by advanced age.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 377-386"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144121521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-01DOI: 10.1016/j.xfss.2025.07.005
R. Clayton Edenfield Ph.D. , Samuel B. Potter B.S. , Krista S. Crow Ph.D. , In Ki Cho Ph.D. , Kristen F. Easley Ph.D. , Nathalia L.M. Lara Ph.D. , Elizabeth S. Waters B.S. , Jason C. Hedges M.D., Ph.D. , Jamie O. Lo M.D. , Ina Dobrinski Ph.D. , Michael Koval Ph.D. , Charles A. Easley Ph.D.
Objective
This study aims to investigate the impact of alcohol on blood-testis barrier (BTB) integrity using a novel in vitro model and to elucidate potential nonhormonal mechanisms underlying alcohologenic reversible azoospermia.
Design
Primary rhesus macaque Sertoli cells were exposed to ethanol to evaluate dose-dependent effects on BTB integrity. Barrier function was assessed through electrical resistance and permeability assays, with recovery evaluated after a 48-hour withdrawal period. Gene expression of Sertoli cells and tight junction markers and cytokine levels associated with barrier degradation were analyzed.
Subjects
Primary rhesus macaque Sertoli cells were used.
Exposure
Clinically relevant concentrations (10, 60, and 100 mM), equivalent to 1–2 drinks to requiring hospitalization, of ethanol were used.
Main Outcome Measures
The main outcome measures were BTB integrity and permeability, assessed via transepithelial electrical resistance and Dye Flux assays, recovery of barrier function after ethanol withdrawal, gene expression changes in Sertoli cells and tight junction markers, and cytokine levels associated with barrier impairment. These metrics evaluated the functional and molecular impacts of in vitro ethanol exposure on the BTB.
Results
Ethanol exposure was associated with a dose-dependent decrease in BTB integrity, with partial recovery observed at lower concentrations (10 and 60 mM) after 48 hours of withdrawal, but not at 100 mM. Additionally, ethanol increased the expression of key Sertoli cells and tight junction genes such as SOX9 and CLDN3, and elevated cytokines associated with barrier degradation at higher concentrations.
Conclusion
Clinically relevant ethanol concentrations reversibly disrupt BTB function through a nonhormonal mechanism, with partial recovery at lower concentrations. These findings provide novel insights into the role of BTB dysfunction in alcohologenic reversible azoospermia.
{"title":"Alcohol alters blood-testis barrier function in an in vitro model","authors":"R. Clayton Edenfield Ph.D. , Samuel B. Potter B.S. , Krista S. Crow Ph.D. , In Ki Cho Ph.D. , Kristen F. Easley Ph.D. , Nathalia L.M. Lara Ph.D. , Elizabeth S. Waters B.S. , Jason C. Hedges M.D., Ph.D. , Jamie O. Lo M.D. , Ina Dobrinski Ph.D. , Michael Koval Ph.D. , Charles A. Easley Ph.D.","doi":"10.1016/j.xfss.2025.07.005","DOIUrl":"10.1016/j.xfss.2025.07.005","url":null,"abstract":"<div><h3>Objective</h3><div>This study aims to investigate the impact of alcohol on blood-testis barrier (BTB) integrity using a novel in vitro model and to elucidate potential nonhormonal mechanisms underlying alcohologenic reversible azoospermia.</div></div><div><h3>Design</h3><div>Primary rhesus macaque Sertoli cells were exposed to ethanol to evaluate dose-dependent effects on BTB integrity. Barrier function was assessed through electrical resistance and permeability assays, with recovery evaluated after a 48-hour withdrawal period. Gene expression of Sertoli cells and tight junction markers and cytokine levels associated with barrier degradation were analyzed.</div></div><div><h3>Subjects</h3><div>Primary rhesus macaque Sertoli cells were used.</div></div><div><h3>Exposure</h3><div>Clinically relevant concentrations (10, 60, and 100 mM), equivalent to 1–2 drinks to requiring hospitalization, of ethanol were used.</div></div><div><h3>Main Outcome Measures</h3><div>The main outcome measures were BTB integrity and permeability, assessed via transepithelial electrical resistance and Dye Flux assays, recovery of barrier function after ethanol withdrawal, gene expression changes in Sertoli cells and tight junction markers, and cytokine levels associated with barrier impairment. These metrics evaluated the functional and molecular impacts of in vitro ethanol exposure on the BTB.</div></div><div><h3>Results</h3><div>Ethanol exposure was associated with a dose-dependent decrease in BTB integrity, with partial recovery observed at lower concentrations (10 and 60 mM) after 48 hours of withdrawal, but not at 100 mM. Additionally, ethanol increased the expression of key Sertoli cells and tight junction genes such as <em>SOX9</em> and <em>CLDN3</em>, and elevated cytokines associated with barrier degradation at higher concentrations.</div></div><div><h3>Conclusion</h3><div>Clinically relevant ethanol concentrations reversibly disrupt BTB function through a nonhormonal mechanism, with partial recovery at lower concentrations. These findings provide novel insights into the role of BTB dysfunction in alcohologenic reversible azoospermia.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 4","pages":"Pages 414-425"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144762551","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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