Pub Date : 2025-05-01Epub Date: 2024-12-28DOI: 10.1016/j.xfss.2024.12.005
Amir Mokhtare Ph.D. , Amirhossein Favakeh M.S. , Philip Xie B.Sc. , Zev Rosenwaks M.D. , Alireza Abbaspourrad Ph.D. , Gianpiero Palermo M.D., Ph.D.
<div><h3>Objective</h3><div>To introduce an innovative noncontact method for denudation process of cumulus-oocyte complexes (COCs) for intracytoplasmic sperm injection (ICSI).</div></div><div><h3>Design</h3><div>We designed and fabricated novel acoustohydrodynamic tweezers (AHTs) to perform contactless denudation and tested them in a mouse model. Cumulus removal efficiency, preimplantation development, and live birth were assessed and compared with those in conventional manual pipetting (MP) denudation.</div></div><div><h3>Subjects</h3><div>Fourteen female B6D2F1/J mice (approximately 4 weeks of age), nine male B6D2F1/J mice (6–12 weeks of age), and 28 CD-1 female mice (approximately 6 weeks of age) were used for experiment.</div></div><div><h3>Exposure</h3><div>We designed a contactless platform for oocyte denudation on the basis of the principle of focalized acoustic waves. We first investigated the acoustic intensity and thermal variability by measuring the surface displacement and temperature with a thermal camera to ensure a safe operation. Cumulus-oocyte complexes were denuded by conventional MP with 40 IU/mL of hyaluronidase serving as control or by AHTs with a reduced amount of hyaluronidase (15 IU/mL). Piezo-ICSI was performed on both experimental and control groups. A triplicate of denudation and insemination experiments was performed. All embryos were monitored in a time-lapse incubator. Embryo developmental rates were compared by the chi-square test. Embryo morphokinetic timing as a continuous variable was compared by 1-way analysis of variance. Embryo transfers were performed on some blastocysts.</div></div><div><h3>Main Outcome Measures</h3><div>The procedural time for each denudation method was measured and compared. Fertilization, embryo development and morphokinetics, pregnancy, and live birth rate were compared between the control and experimental cohorts.</div></div><div><h3>Results</h3><div>Facile noncontact denudation was achieved without any damage to oocyte. Acoustic induced fluidic shear was the main contributor to COC denudation. The average denudation time per oocyte decreased by 46% (15 seconds per oocyte for control vs. 8 seconds per oocyte for AHT) while using a lower concentration of hyaluronidase. Piezo-ICSI on oocytes processed by MP and AHTs resulted in comparable rates of survival (86.1% vs. 85.3%), fertilization (96.7% vs. 94.1%), and blastocyst (88.0% vs. 81.3%). Embryo morphokinetics for both experimental and control cohorts were comparable, showing no impact of sound waves on the embryo development. Eventual delivery rates were also comparable between the MP and AHT cohorts (51.3% vs. 55.4%).</div></div><div><h3>Conclusion</h3><div>Acoustohydrodynamic tweezers are used for contactless removal of the cumulus cells from the COCs before ICSI in an expedited, safe, and reliable manner. Embryo development outcomes confirm their safety and validate their potential for a comprehensive ICSI-on-chip device.</div></div
目的：介绍一种用于卵母细胞胞浆内单精子注射（ICSI）的卵泡卵母细胞复合物（COCs）剥脱过程的创新非接触方法。设计：我们设计并制作了一种新型的声水动力镊子（AHT）来进行非接触剥蚀，并在小鼠模型上进行了测试。评估积云去除效率，植入前发育和活产，并与传统的手动移液剥落进行比较。实验对象：雌性B6D2F1/J小鼠14只（~ 4周龄），雄性B6D2F1/J小鼠9只（6 ~ 12周龄），CD-1雌性小鼠28只（~ 6周龄）进行实验干预和方法：基于聚焦声波原理设计了一种非接触式卵母细胞剥离平台。我们首先通过热像仪测量表面位移和温度来研究声强和热变异性，以确保安全操作。用40 IU/ml透明质酸酶作为对照的常规手动移液（MP）或用减少透明质酸酶量（15 IU/ml）的AHTs剥离COCs。实验组和对照组均行压痕icsi。进行了三次脱毛和人工授精实验。所有胚胎均在延时培养箱中监测。采用卡方检验比较胚胎发育率。胚胎形态动力学时间作为一个连续变量，采用单因素方差分析进行比较。对一些囊胚进行了胚胎移植。主要观察指标：测量并比较各种剥除方法的手术时间。比较对照组和试验组的受精、胚胎发育和形态动力学、妊娠率和活产率。结果：非接触性脱皮容易，无卵母细胞损伤。声波诱导的流体剪切是COC剥蚀的主要原因。当使用较低浓度的透明质酸酶时，每个卵母细胞的平均剥落时间减少了46%（对照组每个卵母细胞15秒，而AHT组每个卵母细胞8秒）。经MP和AHTs处理的卵母细胞的Piezo-ICSI存活率（86.1% vs 85.3%， P=0.80）、受精率（96.7% vs 94.1%， P=0.09）和囊胚率（88.0% vs 81.3%， P=0.06）相当。实验组和对照组的胚胎形态动力学具有可比性，表明声波对胚胎发育没有影响。MP组和AHT组的最终分娩率也具有可比性（51.3% vs 55.4%）。结论：声水动力镊子（AHTs）用于ICSI前COCs积云细胞的非接触清除，快速、安全、可靠。胚胎发育结果证实了它们的安全性，并验证了它们作为综合icsi芯片设备的潜力。
{"title":"A sound approach for ova denudation","authors":"Amir Mokhtare Ph.D. , Amirhossein Favakeh M.S. , Philip Xie B.Sc. , Zev Rosenwaks M.D. , Alireza Abbaspourrad Ph.D. , Gianpiero Palermo M.D., Ph.D.","doi":"10.1016/j.xfss.2024.12.005","DOIUrl":"10.1016/j.xfss.2024.12.005","url":null,"abstract":"<div><h3>Objective</h3><div>To introduce an innovative noncontact method for denudation process of cumulus-oocyte complexes (COCs) for intracytoplasmic sperm injection (ICSI).</div></div><div><h3>Design</h3><div>We designed and fabricated novel acoustohydrodynamic tweezers (AHTs) to perform contactless denudation and tested them in a mouse model. Cumulus removal efficiency, preimplantation development, and live birth were assessed and compared with those in conventional manual pipetting (MP) denudation.</div></div><div><h3>Subjects</h3><div>Fourteen female B6D2F1/J mice (approximately 4 weeks of age), nine male B6D2F1/J mice (6–12 weeks of age), and 28 CD-1 female mice (approximately 6 weeks of age) were used for experiment.</div></div><div><h3>Exposure</h3><div>We designed a contactless platform for oocyte denudation on the basis of the principle of focalized acoustic waves. We first investigated the acoustic intensity and thermal variability by measuring the surface displacement and temperature with a thermal camera to ensure a safe operation. Cumulus-oocyte complexes were denuded by conventional MP with 40 IU/mL of hyaluronidase serving as control or by AHTs with a reduced amount of hyaluronidase (15 IU/mL). Piezo-ICSI was performed on both experimental and control groups. A triplicate of denudation and insemination experiments was performed. All embryos were monitored in a time-lapse incubator. Embryo developmental rates were compared by the chi-square test. Embryo morphokinetic timing as a continuous variable was compared by 1-way analysis of variance. Embryo transfers were performed on some blastocysts.</div></div><div><h3>Main Outcome Measures</h3><div>The procedural time for each denudation method was measured and compared. Fertilization, embryo development and morphokinetics, pregnancy, and live birth rate were compared between the control and experimental cohorts.</div></div><div><h3>Results</h3><div>Facile noncontact denudation was achieved without any damage to oocyte. Acoustic induced fluidic shear was the main contributor to COC denudation. The average denudation time per oocyte decreased by 46% (15 seconds per oocyte for control vs. 8 seconds per oocyte for AHT) while using a lower concentration of hyaluronidase. Piezo-ICSI on oocytes processed by MP and AHTs resulted in comparable rates of survival (86.1% vs. 85.3%), fertilization (96.7% vs. 94.1%), and blastocyst (88.0% vs. 81.3%). Embryo morphokinetics for both experimental and control cohorts were comparable, showing no impact of sound waves on the embryo development. Eventual delivery rates were also comparable between the MP and AHT cohorts (51.3% vs. 55.4%).</div></div><div><h3>Conclusion</h3><div>Acoustohydrodynamic tweezers are used for contactless removal of the cumulus cells from the COCs before ICSI in an expedited, safe, and reliable manner. Embryo development outcomes confirm their safety and validate their potential for a comprehensive ICSI-on-chip device.</div></div","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 118-125"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142904281","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 association between polycystic ovary syndrome (PCOS) and the rate of embryo development, using time-lapse monitoring systems, compared with a control group of women with mechanical (tubal) factor infertility.
Design
A retrospective case-control study conducted in a university-affiliated in vitro fertilization (IVF) unit.
Subjects
Women with PCOS undergoing IVF treatments and those with non-PCOS controls with tubal factor infertility only. Development morphokinetic milestones were compared and analysis of covariance for time to distinct cell number as well as logistic mixed models to determine predictors for embryos over the 75th percentile was performed.
Exposure
Not applicable.
Main Outcome Measures
Embryo development morphokinetic parameters in women with and without PCOS undergoing IVF treatments.
Results
The study included 791 embryos from 115 women, 364 embryos from 52 women with PCOS and 427 embryos from 63 women with non-PCOS controls with tubal factor infertility. The PCOS group was 4 years younger (30.07 ± 6.03 vs. 34.08 ± 4.84 years) and had higher number of oocytes retrieved (16.00 vs. 11.00), mature oocytes (11.00 vs. 7.00) and fertilized oocytes (8.00 vs. 5.00). The PCOS and control groups demonstrated comparable clinical pregnancy rates (55.8% vs. 32.1%), miscarriage rate (12.5% vs. 11.8%), and live birth rate (48.8% vs. 31.2%). Morphokinetic parameters were comparable between the groups. Although age was associated with later time to 5 and 8 discrete cells and start of blastulation (tSB), PCOS was only associated with later tSB, including tSB >75th percentile.
Conclusion
This study demonstrated comparable IVF outcomes in women with PCOS and non-PCOS controls. An analysis of time-lapse monitoring data from these patients showed no evidence that PCOS negatively affects embryonic development rate in women undergoing IVF cycles.
{"title":"Polycystic ovary syndrome and morphokinetic embryonic development: a case-control study evaluating 791 embryos","authors":"Gilad Karavani M.D. , Shira Shapira-Nass M.D. , Natali Schachter-Safrai M.D. , Tal Imbar M.D. , Assaf Ben-Meir M.D.","doi":"10.1016/j.xfss.2025.01.003","DOIUrl":"10.1016/j.xfss.2025.01.003","url":null,"abstract":"<div><h3>Objective</h3><div>To investigate the association between polycystic ovary syndrome (PCOS) and the rate of embryo development, using time-lapse monitoring systems, compared with a control group of women with mechanical (tubal) factor infertility.</div></div><div><h3>Design</h3><div>A retrospective case-control study conducted in a university-affiliated in vitro fertilization (IVF) unit.</div></div><div><h3>Subjects</h3><div>Women with PCOS undergoing IVF treatments and those with non-PCOS controls with tubal factor infertility only. Development morphokinetic milestones were compared and analysis of covariance for time to distinct cell number as well as logistic mixed models to determine predictors for embryos over the 75th percentile was performed.</div></div><div><h3>Exposure</h3><div>Not applicable.</div></div><div><h3>Main Outcome Measures</h3><div>Embryo development morphokinetic parameters in women with and without PCOS undergoing IVF treatments.</div></div><div><h3>Results</h3><div>The study included 791 embryos from 115 women, 364 embryos from 52 women with PCOS and 427 embryos from 63 women with non-PCOS controls with tubal factor infertility. The PCOS group was 4 years younger (30.07 ± 6.03 vs. 34.08 ± 4.84 years) and had higher number of oocytes retrieved (16.00 vs. 11.00), mature oocytes (11.00 vs. 7.00) and fertilized oocytes (8.00 vs. 5.00). The PCOS and control groups demonstrated comparable clinical pregnancy rates (55.8% vs. 32.1%), miscarriage rate (12.5% vs. 11.8%), and live birth rate (48.8% vs. 31.2%). Morphokinetic parameters were comparable between the groups. Although age was associated with later time to 5 and 8 discrete cells and start of blastulation (tSB), PCOS was only associated with later tSB, including tSB >75th percentile.</div></div><div><h3>Conclusion</h3><div>This study demonstrated comparable IVF outcomes in women with PCOS and non-PCOS controls. An analysis of time-lapse monitoring data from these patients showed no evidence that PCOS negatively affects embryonic development rate in women undergoing IVF cycles.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 252-260"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143017331","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 : 2025-05-01Epub Date: 2024-12-28DOI: 10.1016/j.xfss.2024.12.004
Adina Schwartz M.D. , Minnie Malik Ph.D. , Paul Driggers Ph.D. , William H. Catherino M.D., Ph.D.
Objective
To determine if the oral gonadotropin-releasing hormone antagonist relugolix affects leiomyoma extracellular matrix production through the transforming growth factor-beta (TGF-β) pathway.
Design
Laboratory study.
Subjects
None.
Exposure
Exposure of human leiomyoma cells to TGF-β and/or relugolix.
Main Outcome Measures
Production of TGF-β, pSMAD2/3, SMAD2/3, collagen 1A1 (COL1A1), fibronectin (FN1), and versican (VCAN) in treated and untreated leiomyoma cells.
Results
Transforming growth factor-beta 3 production decreased at 24 hours with relugolix 10 nM (0.80 ± 0.09-fold) and 100 nM (0.86 ± 0.06-fold) and at 48 hours with relugolix 1 nM (0.86 ± 0.05-fold) and 100 nM (0.86 ± 0.06-fold). pSMAD2/3 production decreased at 24 hours with relugolix 1 nM (0.71 ± 0.01-fold), 10 nM (0.68 ± 0.01-fold), and 100 nM (0.41 ± 0.10-fold). Compared with relugolix treatment alone at the same concentration, combination treatment at 24 hours resulted in significantly increased COL1A1, FN1, and VCAN production with relugolix 1 nM, 10 nM, and 100 nM. At 48 hours, combination treatment resulted in significantly increased COL1A1, FN1, and VCAN production with relugolix 10 nM and 100 nM.
Conclusion
Relugolix regulated leiomyoma size by decreasing COL1A1, FN1, and VCAN production. This effect is at least partly through the TGF-β pathway.
{"title":"Relugolix reduces leiomyoma extracellular matrix production via the transforming growth factor-beta pathway","authors":"Adina Schwartz M.D. , Minnie Malik Ph.D. , Paul Driggers Ph.D. , William H. Catherino M.D., Ph.D.","doi":"10.1016/j.xfss.2024.12.004","DOIUrl":"10.1016/j.xfss.2024.12.004","url":null,"abstract":"<div><h3>Objective</h3><div>To determine if the oral gonadotropin-releasing hormone antagonist relugolix affects leiomyoma extracellular matrix production through the transforming growth factor-beta (TGF-β) pathway.</div></div><div><h3>Design</h3><div>Laboratory study.</div></div><div><h3>Subjects</h3><div>None.</div></div><div><h3>Exposure</h3><div>Exposure of human leiomyoma cells to TGF-β and/or relugolix.</div></div><div><h3>Main Outcome Measures</h3><div>Production of TGF-β, pSMAD2/3, SMAD2/3, collagen 1A1 (COL1A1), fibronectin (FN1), and versican (VCAN) in treated and untreated leiomyoma cells.</div></div><div><h3>Results</h3><div>Transforming growth factor-beta 3 production decreased at 24 hours with relugolix 10 nM (0.80 ± 0.09-fold) and 100 nM (0.86 ± 0.06-fold) and at 48 hours with relugolix 1 nM (0.86 ± 0.05-fold) and 100 nM (0.86 ± 0.06-fold). pSMAD2/3 production decreased at 24 hours with relugolix 1 nM (0.71 ± 0.01-fold), 10 nM (0.68 ± 0.01-fold), and 100 nM (0.41 ± 0.10-fold). Compared with relugolix treatment alone at the same concentration, combination treatment at 24 hours resulted in significantly increased COL1A1, FN1, and VCAN production with relugolix 1 nM, 10 nM, and 100 nM. At 48 hours, combination treatment resulted in significantly increased COL1A1, FN1, and VCAN production with relugolix 10 nM and 100 nM.</div></div><div><h3>Conclusion</h3><div>Relugolix regulated leiomyoma size by decreasing COL1A1, FN1, and VCAN production. This effect is at least partly through the TGF-β pathway.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 213-220"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142904284","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 : 2025-05-01Epub Date: 2024-12-04DOI: 10.1016/j.xfss.2024.11.002
Nirukshi Samarajeewa Ph.D. , Sophea Heng Ph.D. , Ying Li B.Eng. , Maxine Scelwyn M.D. , Luk J. Rombauts M.D. Ph.D. , Guiying Nie Ph.D.
Objective
To investigate whether endometrial receptivity is affected in patients with endometriosis using podocalyxin (PCX) as a functional biomarker and to study how endometriotic lesions display PCX and the potential pathological implications.
Design
We have previously reported that PCX, an anti-adhesion glycoprotein and barrier protector, is dynamically regulated in the endometrium and acts as a key negative regulator of epithelial receptivity. Early in the cycle both luminal epithelium (LE, lining the endometrial surface) and glandular epithelium (GE, residing within the tissue) strongly express PCX, but in the receptive window, PCX is selectively downregulated in LE, switching the endometrial surface to an adhesive state for embryo attachment/implantation; meanwhile, PCX expression is maintained in GE until postreceptivity. Here, we immuno-stained PCX in endometrial tissues and ectopic lesions biopsied across the menstrual cycle from patients with endometriosis (EOS, n = 41), and compared with endometrium of non-endometriosis controls (non-EOS, n = 55). We further investigated how PCX changes observed in ectopic lesions may influence their adhesive capacity.
Subjects
Women without and with endometriosis.
Exposure
Not applicable.
Main Outcome Measures
The window of endometrial receptivity might be shorter in patients with endometriosis; ectopic sites in addition downregulate PCX cyclically, mirroring the eutopic endometrial cells in preparing for receptivity to increase their adhesion potential.
Results
Endometrial PCX levels were comparable between non-EOS and EOS early in the cycle, and in both groups, PCX is downregulated in LE during the expected window of receptivity; however, in EOS endometrium, PCX is reduced earlier in GE as if the receptive window were shorter. In endometriotic lesions, PCX was detected in endometrial LE- and GE-like cells plus mesothelial cells enveloping peritoneal organs, but PCX was cyclically lost specifically in LE-like cells and reduced in GE-like cells as seen in the eutopic endometrium, which however may increase their adhesion potential to nearby organs (overlaid by mesothelial cells). This speculation was further corroborated in an in vitro model showing endometrial epithelial cells with lower PCX were indeed more adhesive to mesothelial cells.
Conclusion
Endometrial receptivity is subtly affected in patients with endometriosis with a shorter window. Cyclic downregulation of PCX in ectopic sites may have pathological consequences.
{"title":"Receptive window might be shorter in patients with endometriosis and lesions cyclically prepare for implantation","authors":"Nirukshi Samarajeewa Ph.D. , Sophea Heng Ph.D. , Ying Li B.Eng. , Maxine Scelwyn M.D. , Luk J. Rombauts M.D. Ph.D. , Guiying Nie Ph.D.","doi":"10.1016/j.xfss.2024.11.002","DOIUrl":"10.1016/j.xfss.2024.11.002","url":null,"abstract":"<div><h3>Objective</h3><div>To investigate whether endometrial receptivity is affected in patients with endometriosis using podocalyxin (PCX) as a functional biomarker and to study how endometriotic lesions display PCX and the potential pathological implications.</div></div><div><h3>Design</h3><div>We have previously reported that PCX, an anti-adhesion glycoprotein and barrier protector, is dynamically regulated in the endometrium and acts as a key negative regulator of epithelial receptivity. Early in the cycle both luminal epithelium (LE, lining the endometrial surface) and glandular epithelium (GE, residing within the tissue) strongly express PCX, but in the receptive window, PCX is selectively downregulated in LE, switching the endometrial surface to an adhesive state for embryo attachment/implantation; meanwhile, PCX expression is maintained in GE until postreceptivity. Here, we immuno-stained PCX in endometrial tissues and ectopic lesions biopsied across the menstrual cycle from patients with endometriosis (EOS, n = 41), and compared with endometrium of non-endometriosis controls (non-EOS, n = 55). We further investigated how PCX changes observed in ectopic lesions may influence their adhesive capacity.</div></div><div><h3>Subjects</h3><div>Women without and with endometriosis.</div></div><div><h3>Exposure</h3><div>Not applicable.</div></div><div><h3>Main Outcome Measures</h3><div>The window of endometrial receptivity might be shorter in patients with endometriosis; ectopic sites in addition downregulate PCX cyclically, mirroring the eutopic endometrial cells in preparing for receptivity to increase their adhesion potential.</div></div><div><h3>Results</h3><div>Endometrial PCX levels were comparable between non-EOS and EOS early in the cycle, and in both groups, PCX is downregulated in LE during the expected window of receptivity; however, in EOS endometrium, PCX is reduced earlier in GE as if the receptive window were shorter. In endometriotic lesions, PCX was detected in endometrial LE- and GE-like cells plus mesothelial cells enveloping peritoneal organs, but PCX was cyclically lost specifically in LE-like cells and reduced in GE-like cells as seen in the eutopic endometrium, which however may increase their adhesion potential to nearby organs (overlaid by mesothelial cells). This speculation was further corroborated in an in vitro model showing endometrial epithelial cells with lower PCX were indeed more adhesive to mesothelial cells.</div></div><div><h3>Conclusion</h3><div>Endometrial receptivity is subtly affected in patients with endometriosis with a shorter window. Cyclic downregulation of PCX in ectopic sites may have pathological consequences.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 232-241"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142792887","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 : 2025-05-01Epub Date: 2025-01-11DOI: 10.1016/j.xfss.2025.01.002
Lara Houeis M.D. , Graziella van der Plancke B.Sc. , Jen-Yu Wen M.D. , Laurence Dupuy Ph.D. , Elodie Kara Ph.D. , Luciana Cacciottola M.D., Ph.D. , Marie-Christine Maurel Ph.D. , Jacques Donnez M.D., Ph.D. , Marie-Madeleine Dolmans M.D., Ph.D.
Objective
To establish a murine model of chemotherapy-induced diminished ovarian reserve (DOR) and investigate residual fertility after chemotherapy exposure.
Design
Two different chemotherapy protocols were tested to establish a valid DOR model by comparing follicle densities in mice given either protocol or physiological solution. An ovarian stimulation protocol was then selected from among different gonadotropins by counting the number of day 2 embryos obtained from normal mice. Finally, DOR mice were stimulated 5 and 8 weeks after chemotherapy with the chosen gonadotropin protocols, and day 2 embryos were recovered after mating, as was ovarian tissue for further immunohistologic analyses.
Subjects
Seventy-two Naval Medical Research Institute mice.
Exposure
Two different chemotherapy protocols.
Main Outcome Measures
This study compared day 2 embryo counts in both normal and chemotherapy-induced DOR mice. Ovarian histology and morphology were also investigated by follicle counting and classification, as was immunostaining for apoptosis (cleaved caspase-3), activation (phospho-Akt), and proliferation (Ki67).
Results
A dose of 12 mg/kg of busulfan (Bu) + 120 mg/kg of cyclophosphamide (Cy) was chosen to establish the DOR model as it significantly reduced the ovarian reserve compared to both control mice (physiological solution) and the 1.2 mg/kg of Bu + 12 mg/kg of Cy protocol, without depleting it completely. When stimulated with 3.75 IU of Menopur, normal mice produced significantly more embryos than DOR mice given 12 mg/kg of Bu + 120 mg/kg of Cy (41.40 ± 14.74 vs. 23.67 ± 15.55 day 2 embryos). Although the follicle count was statistically diminished after single-dose chemotherapy administration, the remaining follicles did not display any difference in terms of apoptosis, activation, or proliferation rates.
Conclusion
We successfully established a chemotherapy-induced DOR model using 12 mg/kg of Bu + 120 mg/kg of Cy, as evidenced by lower, but not completely depleted, follicle numbers and fewer retrieved embryos. Histologic study of ovarian tissue exposed to DOR-inducing chemotherapy revealed that surviving follicles were of the similar quality as tissue not exposed to chemotherapy.
目的建立化疗引起的卵巢储备功能减退（DOR）的小鼠模型，并研究化疗暴露后的剩余生育能力：设计：测试两种不同的化疗方案，通过比较两种方案中任一方案与生理溶液中小鼠的卵泡密度，建立有效的DOR模型。然后，通过计算正常小鼠第 2 天胚胎的数量，从不同的促性腺激素中选择一种卵巢刺激方案。最后，在化疗后5周和8周用选定的促性腺激素方案刺激DOR小鼠，交配后回收第2天胚胎和卵巢组织，以进一步进行免疫组织学分析：72只海军医学研究所（NMRI）小鼠：比较正常小鼠和化疗诱导的 DOR 小鼠的第 2 天胚胎数。还通过卵泡计数和分类对卵巢组织学和形态学进行了研究，并对凋亡（裂解的caspase-3）、活化（phospho-Akt）和增殖（Ki67）进行了免疫染色：与对照组小鼠（生理溶液）和1.2 mg/kg Bu + 12 mg/kg Cy方案相比，12 mg/kg Busulfan (Bu) + 120 mg/kg cyclophosphamide (Cy)剂量显著降低了卵巢储备功能，但并未完全耗尽，因此被选为建立DOR模型的剂量。在使用 3.75 IU Menopur 的刺激下，正常小鼠产生的胚胎数明显多于使用 12 mg/kg Bu + 120 mg/kg Cy 的 DOR 小鼠（第 2 天胚胎数为 41.40 ± 14.74 对 23.67 ± 15.55）。虽然单剂量化疗后卵泡数量在统计学上有所减少，但剩余卵泡在凋亡、活化或增殖率方面没有任何差异：结论：我们使用12 mg/kg Bu + 120 mg/kg Cy成功建立了化疗诱导的DOR模型，其表现为卵泡数量减少，但并未完全耗竭，取回的胚胎数量也较少。对暴露于DOR诱导化疗的卵巢组织进行的组织学研究显示，存活卵泡的质量与未暴露于化疗的组织相同。
{"title":"Chemotherapy-induced diminished murine ovarian reserve model and impact of low-dose chemotherapy on fertility","authors":"Lara Houeis M.D. , Graziella van der Plancke B.Sc. , Jen-Yu Wen M.D. , Laurence Dupuy Ph.D. , Elodie Kara Ph.D. , Luciana Cacciottola M.D., Ph.D. , Marie-Christine Maurel Ph.D. , Jacques Donnez M.D., Ph.D. , Marie-Madeleine Dolmans M.D., Ph.D.","doi":"10.1016/j.xfss.2025.01.002","DOIUrl":"10.1016/j.xfss.2025.01.002","url":null,"abstract":"<div><h3>Objective</h3><div>To establish a murine model of chemotherapy-induced diminished ovarian reserve (DOR) and investigate residual fertility after chemotherapy exposure.</div></div><div><h3>Design</h3><div>Two different chemotherapy protocols were tested to establish a valid DOR model by comparing follicle densities in mice given either protocol or physiological solution. An ovarian stimulation protocol was then selected from among different gonadotropins by counting the number of day 2 embryos obtained from normal mice. Finally, DOR mice were stimulated 5 and 8 weeks after chemotherapy with the chosen gonadotropin protocols, and day 2 embryos were recovered after mating, as was ovarian tissue for further immunohistologic analyses.</div></div><div><h3>Subjects</h3><div>Seventy-two Naval Medical Research Institute mice.</div></div><div><h3>Exposure</h3><div>Two different chemotherapy protocols.</div></div><div><h3>Main Outcome Measures</h3><div>This study compared day 2 embryo counts in both normal and chemotherapy-induced DOR mice. Ovarian histology and morphology were also investigated by follicle counting and classification, as was immunostaining for apoptosis (cleaved caspase-3), activation (phospho-Akt), and proliferation (Ki67).</div></div><div><h3>Results</h3><div>A dose of 12 mg/kg of busulfan (Bu) + 120 mg/kg of cyclophosphamide (Cy) was chosen to establish the DOR model as it significantly reduced the ovarian reserve compared to both control mice (physiological solution) and the 1.2 mg/kg of Bu + 12 mg/kg of Cy protocol, without depleting it completely. When stimulated with 3.75 IU of Menopur, normal mice produced significantly more embryos than DOR mice given 12 mg/kg of Bu + 120 mg/kg of Cy (41.40 ± 14.74 vs. 23.67 ± 15.55 day 2 embryos). Although the follicle count was statistically diminished after single-dose chemotherapy administration, the remaining follicles did not display any difference in terms of apoptosis, activation, or proliferation rates.</div></div><div><h3>Conclusion</h3><div>We successfully established a chemotherapy-induced DOR model using 12 mg/kg of Bu + 120 mg/kg of Cy, as evidenced by lower, but not completely depleted, follicle numbers and fewer retrieved embryos. Histologic study of ovarian tissue exposed to DOR-inducing chemotherapy revealed that surviving follicles were of the similar quality as tissue not exposed to chemotherapy.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 177-185"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142973627","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 : 2025-05-01Epub Date: 2025-03-25DOI: 10.1016/j.xfss.2025.03.003
Varsha Jain Ph.D. , Emi Hojo M.Sc. , Graham McKillop M.B.Ch.B. , Anca Oniscu M.D. , Yuan Le Ph.D. , Jun Chen Ph.D. , Richard Ehman Ph.D. , Neil Roberts Ph.D. , Hilary O.D. Critchley M.D., D.Sc.
Objective
Magnetic resonance elastography (MRE), a novel imaging technique that allows in vivo measurement of tissue mechanical properties, was used to test the prediction that the stiffness of the uterus may be increased due to fibrotic changes in patients with adenomyosis.
Design
A feasibility study in which a 3-dimensional (3D) MRE imaging protocol was developed to measure the stiffness of the tissues of the uterus.
Subjects
Four patients with suspected adenomyosis and heavy menstrual bleeding diagnosed via transvaginal ultrasound and clinical history and 1 healthy control were recruited. Two patients underwent hysterectomy, and histologic analysis of the tissue samples was performed.
Main Outcome Measures
The stiffness of the whole uterus was obtained by region of interest analysis of the 3D MRE images for the 4 patients and 1 healthy control. In addition, for the 2 patients who underwent hysterectomy, the uterine tissue samples were assessed to determine histologic presence of adenomyosis via hematoxylin and eosin staining, cellular/molecular measures of tissue stiffness (collagen [picrosirius red], α-smooth muscle actin, and e-cadherin), and whether a relationship existed between in vivo assessment of the uterus via 3D MRE and in vitro uterine tissue histology.
Results
3D MRE was successfully used to acquire elastograms for 4 patients with adenomyosis (diffuse, n = 3; focal, n = 1) and 1 healthy control. Calculated global uterine stiffness was higher in women with adenomyosis (2.93 kPa; range, 2.34–3.39 kPa) than in the healthy control (2.04 kPa). Regions of high stiffness on the 3D elastograms reflected adenomyotic changes visualized via conventional magnetic resonance imaging and were correlated with histologic and immunohistochemical markers of tissue stiffness.
Conclusion
3D MRE has the potential to provide non-invasive characterization of changes in the mechanical properties of uterine tissue that is not possible using conventional magnetic resonance imaging or transvaginal ultrasound. Further studies are needed to confirm the efficacy of the 3D MRE protocol for diagnosing adenomyosis.
{"title":"Feasibility study of the application of magnetic resonance elastography to diagnose uterine adenomyosis","authors":"Varsha Jain Ph.D. , Emi Hojo M.Sc. , Graham McKillop M.B.Ch.B. , Anca Oniscu M.D. , Yuan Le Ph.D. , Jun Chen Ph.D. , Richard Ehman Ph.D. , Neil Roberts Ph.D. , Hilary O.D. Critchley M.D., D.Sc.","doi":"10.1016/j.xfss.2025.03.003","DOIUrl":"10.1016/j.xfss.2025.03.003","url":null,"abstract":"<div><h3>Objective</h3><div>Magnetic resonance elastography (MRE), a novel imaging technique that allows in vivo measurement of tissue mechanical properties, was used to test the prediction that the stiffness of the uterus may be increased due to fibrotic changes in patients with adenomyosis.</div></div><div><h3>Design</h3><div>A feasibility study in which a 3-dimensional (3D) MRE imaging protocol was developed to measure the stiffness of the tissues of the uterus.</div></div><div><h3>Subjects</h3><div>Four patients with suspected adenomyosis and heavy menstrual bleeding diagnosed via transvaginal ultrasound and clinical history and 1 healthy control were recruited. Two patients underwent hysterectomy, and histologic analysis of the tissue samples was performed.</div></div><div><h3>Main Outcome Measures</h3><div>The stiffness of the whole uterus was obtained by region of interest analysis of the 3D MRE images for the 4 patients and 1 healthy control. In addition, for the 2 patients who underwent hysterectomy, the uterine tissue samples were assessed to determine histologic presence of adenomyosis via hematoxylin and eosin staining, cellular/molecular measures of tissue stiffness (collagen [picrosirius red], α-smooth muscle actin, and e-cadherin), and whether a relationship existed between in vivo assessment of the uterus via 3D MRE and in vitro uterine tissue histology.</div></div><div><h3>Results</h3><div>3D MRE was successfully used to acquire elastograms for 4 patients with adenomyosis (diffuse, n = 3; focal, n = 1) and 1 healthy control. Calculated global uterine stiffness was higher in women with adenomyosis (2.93 kPa; range, 2.34–3.39 kPa) than in the healthy control (2.04 kPa). Regions of high stiffness on the 3D elastograms reflected adenomyotic changes visualized via conventional magnetic resonance imaging and were correlated with histologic and immunohistochemical markers of tissue stiffness.</div></div><div><h3>Conclusion</h3><div>3D MRE has the potential to provide non-invasive characterization of changes in the mechanical properties of uterine tissue that is not possible using conventional magnetic resonance imaging or transvaginal ultrasound. Further studies are needed to confirm the efficacy of the 3D MRE protocol for diagnosing adenomyosis.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 242-251"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143733488","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 : 2025-05-01Epub Date: 2025-02-17DOI: 10.1016/j.xfss.2025.02.001
Ramanaiah Mamillapalli Ph.D., Rebecca Slutzky B.A., Anjali Mangla B.A., Nimisha Gawde M.D., Hugh S. Taylor M.D.
Objective
To determine if microRNAs that are altered in the circulation of women with endometriosis affect metabolic gene expression in hepatic cells.
Design
In vitro study.
Subjects
Deidentified tissue from women with endometriosis.
Exposure
MicroRNAs were used to induce or suppress target genes in hepatic cells.
Main Outcome Measures
Effect of the microRNAs that are aberrantly expressed in endometriosis on hepatic cell gene expression using quantitative polymerase chain reaction.
Results
Prior microarray studies on the serum of women with endometriosis showed differential expression of microRNAs miR-Let-7b, miR-125b-5p, miR-150-5p, and miR-3613-5p. Bioinformatic analyses revealed that these microRNAs have predicted binding sites in multiple genes involved in liver metabolism. Transfection of these miRs in HepG2 cells followed by real-time quantitative polymerase chain reaction showed that miR-Let-7b mimic increased the expression of Igfbp1 by 8-fold and reduced the expression of Mrc1 by 3.2-fold, whereas its inhibitor reduced Igfbp1 by 2.8-fold and increased Mrc1 by 5.2-fold. MiR-3613-5p mimic reduced the expression of Cyp2r1 by 2.2-fold and Mrc1 by 4-fold. MiR-125b-5p mimic increased the expression of Fabp4 by 4.1-fold, whereas miR-150-5p mimic increased the expression of Mrc1 by 1.8-fold and Cyp2r1 by 2.5-fold. Inhibitors of both miR-125b-5p and miR-150-5p did not show any effect on any of the genes.
Conclusion
Circulating microRNAs, known to be aberrant in endometriosis-regulated hepatic gene expression, likely contribute to the metabolic defects seen in this disease. Treatment with miR-Let-7b and miR-3613-5p, which are downregulated in endometriosis, reversed the effect of endometriosis on the expression of IGFBP1, MRC1, and CYP2r1 genes. Therefore, miR-Let-7b and miR-3613-5p may be novel candidate therapies for endometriosis, potentially correcting the metabolic changes seen in patients with endometriosis.
{"title":"Effect of endometriosis-linked microRNAs on hepatic gene expression","authors":"Ramanaiah Mamillapalli Ph.D., Rebecca Slutzky B.A., Anjali Mangla B.A., Nimisha Gawde M.D., Hugh S. Taylor M.D.","doi":"10.1016/j.xfss.2025.02.001","DOIUrl":"10.1016/j.xfss.2025.02.001","url":null,"abstract":"<div><h3>Objective</h3><div>To determine if microRNAs that are altered in the circulation of women with endometriosis affect metabolic gene expression in hepatic cells.</div></div><div><h3>Design</h3><div>In vitro study.</div></div><div><h3>Subjects</h3><div>Deidentified tissue from women with endometriosis.</div></div><div><h3>Exposure</h3><div>MicroRNAs were used to induce or suppress target genes in hepatic cells.</div></div><div><h3>Main Outcome Measures</h3><div>Effect of the microRNAs that are aberrantly expressed in endometriosis on hepatic cell gene expression using quantitative polymerase chain reaction.</div></div><div><h3>Results</h3><div>Prior microarray studies on the serum of women with endometriosis showed differential expression of microRNAs miR-Let-7b, miR-125b-5p, miR-150-5p, and miR-3613-5p. Bioinformatic analyses revealed that these microRNAs have predicted binding sites in multiple genes involved in liver metabolism. Transfection of these miRs in HepG2 cells followed by real-time quantitative polymerase chain reaction showed that miR-Let-7b mimic increased the expression of <em>Igfbp1</em> by 8-fold and reduced the expression of <em>Mrc1</em> by 3.2-fold, whereas its inhibitor reduced <em>Igfbp1</em> by 2.8-fold and increased <em>Mrc1</em> by 5.2-fold. MiR-3613-5p mimic reduced the expression of <em>Cyp2r1</em> by 2.2-fold and <em>Mrc1</em> by 4-fold. MiR-125b-5p mimic increased the expression of <em>Fabp4</em> by 4.1-fold, whereas miR-150-5p mimic increased the expression of <em>Mrc1</em> by 1.8-fold and <em>Cyp2r1</em> by 2.5-fold. Inhibitors of both miR-125b-5p and miR-150-5p did not show any effect on any of the genes.</div></div><div><h3>Conclusion</h3><div>Circulating microRNAs, known to be aberrant in endometriosis-regulated hepatic gene expression, likely contribute to the metabolic defects seen in this disease. Treatment with miR-Let-7b and miR-3613-5p, which are downregulated in endometriosis, reversed the effect of endometriosis on the expression of <em>IGFBP1</em>, <em>MRC1,</em> and <em>CYP2r1</em> genes. Therefore, miR-Let-7b and miR-3613-5p may be novel candidate therapies for endometriosis, potentially correcting the metabolic changes seen in patients with endometriosis.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 221-231"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143461057","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 : 2025-05-01Epub Date: 2025-03-28DOI: 10.1016/j.xfss.2025.03.006
Javier Guerrero-Sánchez Ms.C. , Andrea Fernández-Toribio Ms.C. , Beatriz Galiano-Cogolludo Ms.C. , Tania García-Martínez Ph.D. , Lucía Mendoza Ph.D. , Gonzalo Fernández-Blanco Ms.C. , Jesús Ramos-Membrive Ms.C. , Joana Fidalgo Ph.D. , Lionel Matthys Ms.C. , José Antonio Horcajadas Ph.D. , Santiago Munné Ph.D. , Pablo Bermejo-Álvarez D.V.M., Ph.D.
Objective
To test the developmental ability of murine zygotes vitrified using a novel vitrification device and microfluidic chip (DaVitri, Overture Life).
Design
Murine zygotes were randomly allocated to 2 groups; one was vitrified using the vitrification device, and the other was following a conventional manual protocol.
Subjects
Murine zygotes obtained in vivo.
Exposure
Automatic vitrification was achieved by a linear exposure to cryoprotectants (CPAs) using the DaVitri device. Manual vitrification was conducted using Kitazato kit.
Main Outcome Measures
Morphokinetic behavior of the zygotes during the exposure to CPAs analyzed by microscopy, developmental rates after thawing, lineage development at the blastocyst stage assessed by immunohistochemistry and light-structured fluorescent microscopy, and survival rates and pup weight after embryo transfer.
Results
Automated vitrification led to a gradual reduction in zygote volume during the equilibration steps preceding ultrafast cooling in liquid nitrogen, as opposed to the conventional manual protocol where sharp changes in zygote volume were observed as a result of exposure to static concentrations of CPAs. Survival rates of the automated procedure were comparable to those of the manual protocol, resulting in ∼95% blastocyst formation rates. Developmental analysis of the resulting blastocysts revealed comparable numbers of total, trophectoderm, and inner cell mass numbers in blastocysts developed from zygotes vitrified under the manual and automated protocols. No differences were found in survival to term or pup weight a D1 or D21.
Conclusion
Automated vitrification using DaVitri device diminished the osmotic stress caused by exposure to CPAs during the equilibration steps and resulted in comparable developmental competence in terms of development to blastocysts, lineage segregation, and survival to term.
{"title":"Kinetics of cell shrinkage and developmental competence of mouse zygotes vitrified following conventional or automated (DaVitri) protocols","authors":"Javier Guerrero-Sánchez Ms.C. , Andrea Fernández-Toribio Ms.C. , Beatriz Galiano-Cogolludo Ms.C. , Tania García-Martínez Ph.D. , Lucía Mendoza Ph.D. , Gonzalo Fernández-Blanco Ms.C. , Jesús Ramos-Membrive Ms.C. , Joana Fidalgo Ph.D. , Lionel Matthys Ms.C. , José Antonio Horcajadas Ph.D. , Santiago Munné Ph.D. , Pablo Bermejo-Álvarez D.V.M., Ph.D.","doi":"10.1016/j.xfss.2025.03.006","DOIUrl":"10.1016/j.xfss.2025.03.006","url":null,"abstract":"<div><h3>Objective</h3><div>To test the developmental ability of murine zygotes vitrified using a novel vitrification device and microfluidic chip (DaVitri, Overture Life).</div></div><div><h3>Design</h3><div>Murine zygotes were randomly allocated to 2 groups; one was vitrified using the vitrification device, and the other was following a conventional manual protocol.</div></div><div><h3>Subjects</h3><div>Murine zygotes obtained in vivo.</div></div><div><h3>Exposure</h3><div>Automatic vitrification was achieved by a linear exposure to cryoprotectants (CPAs) using the DaVitri device. Manual vitrification was conducted using Kitazato kit.</div></div><div><h3>Main Outcome Measures</h3><div>Morphokinetic behavior of the zygotes during the exposure to CPAs analyzed by microscopy, developmental rates after thawing, lineage development at the blastocyst stage assessed by immunohistochemistry and light-structured fluorescent microscopy, and survival rates and pup weight after embryo transfer.</div></div><div><h3>Results</h3><div>Automated vitrification led to a gradual reduction in zygote volume during the equilibration steps preceding ultrafast cooling in liquid nitrogen, as opposed to the conventional manual protocol where sharp changes in zygote volume were observed as a result of exposure to static concentrations of CPAs. Survival rates of the automated procedure were comparable to those of the manual protocol, resulting in ∼95% blastocyst formation rates. Developmental analysis of the resulting blastocysts revealed comparable numbers of total, trophectoderm, and inner cell mass numbers in blastocysts developed from zygotes vitrified under the manual and automated protocols. No differences were found in survival to term or pup weight a D1 or D21.</div></div><div><h3>Conclusion</h3><div>Automated vitrification using DaVitri device diminished the osmotic stress caused by exposure to CPAs during the equilibration steps and resulted in comparable developmental competence in terms of development to blastocysts, lineage segregation, and survival to term.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 186-194"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143756321","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 : 2025-05-01Epub Date: 2025-02-25DOI: 10.1016/j.xfss.2025.02.007
Pamela B. Parker M.D., M.P.H. , Melinda J. Murphy B.S. , Sweta Ravisankar Ph.D. , Shawn L. Chavez Ph.D. , Jon D. Hennebold Ph.D.
Objective
To determine the effect of an obesogenic Western-style diet and hyperandrogenemia on ovarian outcomes.
Design
Experimental, controlled animal study.
Subjects
Post-pubertal rhesus macaque females.
Exposure
A Western-style diet (WSD) (WSD: 36% fat, 45% carbohydrate, 18% protein) combined with exogenously administered testosterone (T) vs. a standard chow diet (control; 15% fat, 59% carbohydrate, 27% protein). Animals underwent controlled ovarian stimulations to assess ovarian follicle development.
Main Outcome Measures
Cycle length, the proportion of ovulatory cycles, and daily levels of estradiol (E2), progesterone, antimüllerian hormone, luteinizing hormone, and follicle-stimulating hormone were compared between control and T+WSD groups through one menstrual cycle. Follicular fluid was assessed for cytokine and steroid content, and retrieved oocytes were evaluated for meiotic maturation and underwent in vitro fertilization. Granulosa cells were analyzed for differential gene expression. Ovaries were removed in early luteal phase (4 days post midcycle estradiol surge) and analyzed for morphological differences.
Results
The T+WSD group demonstrated significantly decreased luteal progesterone levels. We found no differences in cycle length, proportion of ovulatory cycles, day of E2 surge, total E2 synthesis, follicle-stimulating hormone, luteinizing hormone or antimüllerian hormone. Analysis of follicular fluid retrieved from animals undergoing an ovarian stimulation protocol revealed increased vascular endothelial growth factor-A, elevated cortisol:cortisone ratio, and increased testosterone and progesterone levels in the treatment group. Granulosa cells from T+WSD demonstrated significantly up-regulated or down-regulated genes relative to controls, including those related to cell differentiation and migration. The ovarian morphology of treatment animals demonstrated enlarged cystic follicles reminiscent of polycystic ovaries.
Conclusion
Similar to prior studies assessing long-term exposure (5–6 years) to T+WSD in female rhesus macaques beginning before menarche, a 1-year T+WSD treatment in adult, regularly cycling females led to reduced luteal phase progesterone levels and polycystic ovarian morphology. Additionally, short-term T+WSD exposure resulted in altered granulosa cell gene expression. Although 1 year of T+WSD exposure leads to altered luteal progesterone, follicular fluid steroid and cytokine content, and granulosa cell gene expression changes, insults of longer duration are required to exert additional negative effects on ovarian function.
目的：探讨致肥性西式饮食和高雄激素血症对卵巢结局的影响。设计：实验对照动物研究对象：青春期后恒河猴雌性暴露：西式饮食（T+WSD: 36%脂肪，45%碳水化合物，18%蛋白质）结合外源性睾酮与标准鼠粮(对照组，CTRL；15%脂肪，59%碳水化合物，27%蛋白质)。实验动物接受受控卵巢刺激以评估卵泡发育。主要观察指标：比较按CTRL组和按T+WSD组在一个月经周期内的周期长度、排卵周期比例、每日雌二醇（E2）、黄体酮（P4）、AMH、黄体生成素（LH）、促卵泡激素（FSH）水平。对卵泡液进行细胞因子和类固醇含量评估，对提取的卵母细胞进行减数分裂成熟评估，并进行体外受精。对颗粒细胞进行差异基因表达分析。在黄体早期（中期雌二醇激增后4天）切除卵巢，分析形态学差异。结果：T+WSD组黄体P4水平明显降低。我们发现在周期长度、排卵周期比例、E2激增天数、总E2合成、FSH、LH或AMH方面没有差异。从接受卵巢刺激方案的动物中提取的卵泡液分析显示，治疗组血管内皮生长因子- a （VEGFA）升高，皮质醇：可的松比值升高，睾酮和孕酮水平升高。与先前评估雌性恒河猴月经初潮前长期（5-6年）暴露于T+WSD的研究类似，在成年、定期循环的雌性恒河猴中，1年的T+WSD治疗导致黄体期黄体酮水平降低和多囊卵巢形态。此外，短期T+WSD暴露导致颗粒细胞基因表达改变。虽然1年的T+WSD暴露会导致黄体黄体酮、卵泡液类固醇和细胞因子含量的改变，以及颗粒细胞基因表达的变化，但需要更长的时间才能对卵巢功能产生额外的负面影响。
{"title":"Impact of a short-term Western-style diet and hyperandrogenism on adult rhesus macaque ovarian function","authors":"Pamela B. Parker M.D., M.P.H. , Melinda J. Murphy B.S. , Sweta Ravisankar Ph.D. , Shawn L. Chavez Ph.D. , Jon D. Hennebold Ph.D.","doi":"10.1016/j.xfss.2025.02.007","DOIUrl":"10.1016/j.xfss.2025.02.007","url":null,"abstract":"<div><h3>Objective</h3><div>To determine the effect of an obesogenic Western-style diet and hyperandrogenemia on ovarian outcomes.</div></div><div><h3>Design</h3><div>Experimental, controlled animal study.</div></div><div><h3>Subjects</h3><div>Post-pubertal rhesus macaque females.</div></div><div><h3>Exposure</h3><div>A Western-style diet (WSD) (WSD: 36% fat, 45% carbohydrate, 18% protein) combined with exogenously administered testosterone (T) vs. a standard chow diet (control; 15% fat, 59% carbohydrate, 27% protein). Animals underwent controlled ovarian stimulations to assess ovarian follicle development.</div></div><div><h3>Main Outcome Measures</h3><div>Cycle length, the proportion of ovulatory cycles, and daily levels of estradiol (E2), progesterone, antimüllerian hormone, luteinizing hormone, and follicle-stimulating hormone were compared between control and T+WSD groups through one menstrual cycle. Follicular fluid was assessed for cytokine and steroid content, and retrieved oocytes were evaluated for meiotic maturation and underwent in vitro fertilization. Granulosa cells were analyzed for differential gene expression. Ovaries were removed in early luteal phase (4 days post midcycle estradiol surge) and analyzed for morphological differences.</div></div><div><h3>Results</h3><div>The T+WSD group demonstrated significantly decreased luteal progesterone levels. We found no differences in cycle length, proportion of ovulatory cycles, day of E2 surge, total E2 synthesis, follicle-stimulating hormone, luteinizing hormone or antimüllerian hormone. Analysis of follicular fluid retrieved from animals undergoing an ovarian stimulation protocol revealed increased vascular endothelial growth factor-A, elevated cortisol:cortisone ratio, and increased testosterone and progesterone levels in the treatment group. Granulosa cells from T+WSD demonstrated significantly up-regulated or down-regulated genes relative to controls, including those related to cell differentiation and migration. The ovarian morphology of treatment animals demonstrated enlarged cystic follicles reminiscent of polycystic ovaries.</div></div><div><h3>Conclusion</h3><div>Similar to prior studies assessing long-term exposure (5–6 years) to T+WSD in female rhesus macaques beginning before menarche, a 1-year T+WSD treatment in adult, regularly cycling females led to reduced luteal phase progesterone levels and polycystic ovarian morphology. Additionally, short-term T+WSD exposure resulted in altered granulosa cell gene expression. Although 1 year of T+WSD exposure leads to altered luteal progesterone, follicular fluid steroid and cytokine content, and granulosa cell gene expression changes, insults of longer duration are required to exert additional negative effects on ovarian function.</div></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 141-151"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143525341","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 : 2025-05-01Epub Date: 2025-01-19DOI: 10.1016/j.xfss.2025.01.004
Papri Sarkar M.D. , Monica Moore M.Sc , Asli Ozmen PhD , Busra Cetinkaya-Un Ph.D , Vitko Julie M.D. , Anthony N. Imudia M.D , Charles J. Lockwood M.D. , Umit A. Kayisli Ph.D. , Ozlem Guzeloglu-Kayisli Ph.D.
<div><h3>Objective</h3><div>To study the relationship between FK506-binding protein 51 (FKBP51) and ovarian aging and/or diminished ovarian reserve (DOR) in human ovaries by comparing FKBP51 levels in granulosa cells (GCs) and cumulus cells (CCs), collected during controlled ovarian stimulation (COS) from women of advanced reproductive age and/or with a diagnosis of DOR with that of young women with normal ovarian reserve. To explore the association between increased FKBP51 expression and human ovarian aging further, expression of FKBP51 was compared in ovarian stroma of postmenopausal vs. premenopausal women. Lastly, this relation was further queried by comparing ovarian expression of several collagen genes as markers of ovarian fibrosis in 14-month-old wild-type (<em>Fkbp5</em><sup><em>+/+</em></sup>) and <em>Fkbp5</em> knockout (<em>Fkbp5</em><sup><em>−/−</em></sup>) mice.</div></div><div><h3>Design</h3><div>Laboratory-based experimental study.</div></div><div><h3>Subjects</h3><div>Samples collected included follicular fluid, CCs, GCs, and serum from group 1: young women with normal ovarian reserve (<35 years; n = 12); group 2: DOR (antimüllerian hormone <1 ng/mL; n = 10); and group 3: women of advanced age with normal ovarian reserve (>37 years; n = 8). Ovarian stromal tissues obtained from surgical specimen of post-menopausal (50–65 years; n = 6) and pre-menopausal (18–30 years; n = 6). Ovarian tissues from 14-month-old <em>Fkbp5</em><sup><em>+/+</em></sup> <em>and Fkbp5</em><sup><em>−/−</em></sup> mice. All the experiments were performed at an academic-affiliated assisted reproductive technology unit/laboratory.</div></div><div><h3>Exposure</h3><div>Comparison of FKBP51 expression in GCs and CCs from women undergoing COS, ovarian stromal tissue from pre- and post-menopausal women, and ovarian tissue from aged <em>Fkbp5</em><sup><em>+/+</em></sup> <em>and Fkbp5</em><sup><em>−/−</em></sup> mice.</div></div><div><h3>Main Outcome Measures</h3><div>(1) Level of FKBP51 in human GCs and CCs, collected during COS by performing real-time quantitative polymerase chain reaction (qPCR). (2) Immunohistochemistry to detect FKBP51 levels and Picrosirius Red staining to detect collagen deposition in human ovarian stromal tissue. (3) Real-time qPCR to compare expression levels of several collagen genes in <em>Fkbp5</em><sup><em>+/+</em></sup> and <em>Fkbp5</em><sup><em>−/−</em></sup> old mice ovaries. Serum and follicular fluid levels of transforming growth factor β1, and soluble endoglin measured by enzyme-linked immunosorbent assay.</div></div><div><h3>Results</h3><div>Immunohistochemistry revealed that FKBP51 histologic score levels in ovarian stromal tissue were significantly higher in postmenopausal vs. premenopausal women (mean ± SEM, 160.52 ± 17.75 vs. 120.67 ± 14.33; <em>P</em>=.002). Stronger Picrosirius Red staining, suggestive of fibrosis, was seen in ovarian stromal tissue of postmenopausal vs. premenopausal women (54.06 ± 6.94 vs. 37.5
{"title":"Enhanced ovarian FKBP51 expression is associated with ovarian aging: a molecular insight for age-related fertility in women","authors":"Papri Sarkar M.D. , Monica Moore M.Sc , Asli Ozmen PhD , Busra Cetinkaya-Un Ph.D , Vitko Julie M.D. , Anthony N. Imudia M.D , Charles J. Lockwood M.D. , Umit A. Kayisli Ph.D. , Ozlem Guzeloglu-Kayisli Ph.D.","doi":"10.1016/j.xfss.2025.01.004","DOIUrl":"10.1016/j.xfss.2025.01.004","url":null,"abstract":"<div><h3>Objective</h3><div>To study the relationship between FK506-binding protein 51 (FKBP51) and ovarian aging and/or diminished ovarian reserve (DOR) in human ovaries by comparing FKBP51 levels in granulosa cells (GCs) and cumulus cells (CCs), collected during controlled ovarian stimulation (COS) from women of advanced reproductive age and/or with a diagnosis of DOR with that of young women with normal ovarian reserve. To explore the association between increased FKBP51 expression and human ovarian aging further, expression of FKBP51 was compared in ovarian stroma of postmenopausal vs. premenopausal women. Lastly, this relation was further queried by comparing ovarian expression of several collagen genes as markers of ovarian fibrosis in 14-month-old wild-type (<em>Fkbp5</em><sup><em>+/+</em></sup>) and <em>Fkbp5</em> knockout (<em>Fkbp5</em><sup><em>−/−</em></sup>) mice.</div></div><div><h3>Design</h3><div>Laboratory-based experimental study.</div></div><div><h3>Subjects</h3><div>Samples collected included follicular fluid, CCs, GCs, and serum from group 1: young women with normal ovarian reserve (<35 years; n = 12); group 2: DOR (antimüllerian hormone <1 ng/mL; n = 10); and group 3: women of advanced age with normal ovarian reserve (>37 years; n = 8). Ovarian stromal tissues obtained from surgical specimen of post-menopausal (50–65 years; n = 6) and pre-menopausal (18–30 years; n = 6). Ovarian tissues from 14-month-old <em>Fkbp5</em><sup><em>+/+</em></sup> <em>and Fkbp5</em><sup><em>−/−</em></sup> mice. All the experiments were performed at an academic-affiliated assisted reproductive technology unit/laboratory.</div></div><div><h3>Exposure</h3><div>Comparison of FKBP51 expression in GCs and CCs from women undergoing COS, ovarian stromal tissue from pre- and post-menopausal women, and ovarian tissue from aged <em>Fkbp5</em><sup><em>+/+</em></sup> <em>and Fkbp5</em><sup><em>−/−</em></sup> mice.</div></div><div><h3>Main Outcome Measures</h3><div>(1) Level of FKBP51 in human GCs and CCs, collected during COS by performing real-time quantitative polymerase chain reaction (qPCR). (2) Immunohistochemistry to detect FKBP51 levels and Picrosirius Red staining to detect collagen deposition in human ovarian stromal tissue. (3) Real-time qPCR to compare expression levels of several collagen genes in <em>Fkbp5</em><sup><em>+/+</em></sup> and <em>Fkbp5</em><sup><em>−/−</em></sup> old mice ovaries. Serum and follicular fluid levels of transforming growth factor β1, and soluble endoglin measured by enzyme-linked immunosorbent assay.</div></div><div><h3>Results</h3><div>Immunohistochemistry revealed that FKBP51 histologic score levels in ovarian stromal tissue were significantly higher in postmenopausal vs. premenopausal women (mean ± SEM, 160.52 ± 17.75 vs. 120.67 ± 14.33; <em>P</em>=.002). Stronger Picrosirius Red staining, suggestive of fibrosis, was seen in ovarian stromal tissue of postmenopausal vs. premenopausal women (54.06 ± 6.94 vs. 37.5","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"6 2","pages":"Pages 152-163"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143017305","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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