Pub Date : 2024-05-01DOI: 10.1016/j.xfss.2024.03.001
Leonardo Augusto Lombardi Ph.D. , Leandro Sabará Mattos M.Sc. , Ana Paula Espindula Ph.D. , Ricardo Santos Simões Ph.D. , Gisela Rodrigues da Silva Sasso Ph.D. , Manuel de Jesus Simões Ph.D. , José Maria Soares-Jr Ph.D. , Rinaldo Florencio-Silva Ph.D.
Objective
To study the combined and isolated effects of melatonin and metformin in the ovarian tissue of rats with PCOS.
Design
Experimental study using a rat model of PCOS induced by continuous light exposure.
Intervention(s)
Forty adult female rats were divided into 5 groups: physiological estrus phase (Sham); permanente estrus with PCOS induced by continuous lighting exposure for 60 consecutive days (control); with PCOS treated with melatonin; with PCOS treated with metformin; with PCOS treated with melatonin + metformin. After 60 days of treatments, all rats were killed, and ovaries were collected and processed for paraffin-embedding. Formalin-fixed paraffin-embedded sections were stained with hematoxylin and eosin or subjected to immunohistochemistry for proliferation (Ki-67) and apoptosis (cleaved caspase 3) detection markers.
Number of corpus luteum and ovarian cysts, number of ovarian follicles (primary and antral follicles), number of interstitial cells, percentage of ovarian follicles (primary and antral follicles), and of interstitial cells immunostained to cleaved caspase-3 and Ki-67.
Results
Absence of corpus luteum, a higher number of cysts, and increased nuclear volume and area of interstitial cells, along with a decrease in primary and antral follicle numbers, were noticed in the control group compared with the Sham group. Melatonin and metformin treatments attenuated these effects, although the combined treatment did not mitigate the increased number of cysts and ovaries induced by PCOS. An increase in theca interna cell apoptosis was observed in the control group, whereas melatonina and metformin treatments reduced it significantly. A higher percentage of caspase-3-immunostained granulosa cells was noted in the Sham and all treated groups compared with the control group; no aditive effects on ovarian cell apoptosis were observed in the combined treatment. The percentage of Ki-67- immunostained granulosa cells was significantly higher in the control group compared with the Sham group. However, the combined treatment, not melatonin and metformin alone, mitigated this effect. A higher percentage of Ki-67-immunostained interstitial cells was observed in all treated groups compared with the Sham and control groups, whereas no additive effects in that immunoreactivity were observed in the combined treatment.
Conclusions
Melatonin and metformin may improve ovarian function in rats with PCOS. The combined melatonin and metformin treatment is more effective in attenuating excessive granulosa cell proliferation, but it is not more effective in improving ovarian function than these drugs applied alone in
{"title":"Effects of melatonin and metformin on the ovaries of rats with polycystic ovary syndrome","authors":"Leonardo Augusto Lombardi Ph.D. , Leandro Sabará Mattos M.Sc. , Ana Paula Espindula Ph.D. , Ricardo Santos Simões Ph.D. , Gisela Rodrigues da Silva Sasso Ph.D. , Manuel de Jesus Simões Ph.D. , José Maria Soares-Jr Ph.D. , Rinaldo Florencio-Silva Ph.D.","doi":"10.1016/j.xfss.2024.03.001","DOIUrl":"10.1016/j.xfss.2024.03.001","url":null,"abstract":"<div><h3>Objective</h3><p>To study the combined and isolated effects of melatonin and metformin in the ovarian tissue of rats with PCOS.</p></div><div><h3>Design</h3><p>Experimental study using a rat model of PCOS induced by continuous light exposure.</p></div><div><h3>Intervention(s)</h3><p>Forty adult female rats were divided into 5 groups: physiological estrus phase (Sham); permanente estrus with PCOS induced by continuous lighting exposure for 60 consecutive days (control); with PCOS treated with melatonin; with PCOS treated with metformin; with PCOS treated with melatonin + metformin. After 60 days of treatments, all rats were killed, and ovaries were collected and processed for paraffin-embedding. Formalin-fixed paraffin-embedded sections were stained with hematoxylin and eosin or subjected to immunohistochemistry for proliferation (Ki-67) and apoptosis (cleaved caspase 3) detection markers.</p></div><div><h3>Setting</h3><p>Federal University of São Paulo, Brazil.</p></div><div><h3>Animals</h3><p>Forty adult female Wistar rats (<em>Rattus norvegicus albinus</em>).</p></div><div><h3>Main Outcome Measure(s)</h3><p>Number of corpus luteum and ovarian cysts, number of ovarian follicles (primary and antral follicles), number of interstitial cells, percentage of ovarian follicles (primary and antral follicles), and of interstitial cells immunostained to cleaved caspase-3 and Ki-67.</p></div><div><h3>Results</h3><p>Absence of corpus luteum, a higher number of cysts, and increased nuclear volume and area of interstitial cells, along with a decrease in primary and antral follicle numbers, were noticed in the control group compared with the Sham group. Melatonin and metformin treatments attenuated these effects, although the combined treatment did not mitigate the increased number of cysts and ovaries induced by PCOS. An increase in theca interna cell apoptosis was observed in the control group, whereas melatonina and metformin treatments reduced it significantly. A higher percentage of caspase-3-immunostained granulosa cells was noted in the Sham and all treated groups compared with the control group; no aditive effects on ovarian cell apoptosis were observed in the combined treatment. The percentage of Ki-67- immunostained granulosa cells was significantly higher in the control group compared with the Sham group. However, the combined treatment, not melatonin and metformin alone, mitigated this effect. A higher percentage of Ki-67-immunostained interstitial cells was observed in all treated groups compared with the Sham and control groups, whereas no additive effects in that immunoreactivity were observed in the combined treatment.</p></div><div><h3>Conclusions</h3><p>Melatonin and metformin may improve ovarian function in rats with PCOS. The combined melatonin and metformin treatment is more effective in attenuating excessive granulosa cell proliferation, but it is not more effective in improving ovarian function than these drugs applied alone in ","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 2","pages":"Pages 204-211"},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140133378","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 adverse effects of phthalate-induced ovarian toxicity on the ovarian reserve and ovarian function. To assess whether the accumulation of higher levels of selected phthalate metabolites in the follicular fluid (FF) of Indian women undergoing intracytoplasmic sperm injection (ICSI) was associated with a decline in their antral follicle count (AFC) and/or serum antimüllerian hormone (AMH) levels, suggesting a negative impact on the ovarian reserve. To evaluate the effects of follicular phthalate metabolites on peak serum estradiol (E2) levels and the total number of oocytes and mature metaphase II (MII) stage oocytes retrieved to assess the impact of phthalate toxicity on ovarian function.
Design
A subanalysis of an ongoing prospective cohort study was conducted to examine the association between the levels of six phthalate metabolites, namely, mono-n-butyl phthalate (MBP), mono-ethyl phthalate (MEP), mono-isononyl phthalate (MiNP), mono-isodecyl phthalate (MiDP), mono(2-ethyl-5-oxohexyl) phthalate, and mono(2-ethyl-5-hydroxyhexyl) phthalate, in the FF of Indian women undergoing ICSI and their ovarian reserve markers (AFC and serum AMH levels). To investigate the association of these follicular phthalate metabolite levels with the peak E2 levels and the total number of oocytes and number of MII stage oocytes retrieved.
Setting
In vitro fertilization center in a referral hospital in India.
Patient(s)
A total of 245 consenting Indian women who had undergone oocyte retrieval between April 2017 and mid-March 2020 were included. Each woman contributed one FF sample to the study. This was screened for six phthalate metabolites. The samples were collected before the coronavirus disease 2019 pandemic.
Intervention(s)
Using liquid chromatography-tandem mass spectrometry, the total levels of six phthalate metabolites were quantified in the FF of 245 women. Using linear regression models that were unadjusted and adjusted for maternal age and body mass index (BMI), we evaluated the association between the follicular metabolites in these women and their AFC, serum AMH levels, peak E2 levels, total number of oocytes, and MII stage oocytes.
Main Outcome Measure(s)
To evaluate the impact of phthalate-induced ovarian toxicity on the ovarian reserve and ovarian function in Indian women undergoing ICSI by studying their accumulated levels in their FF.
Result(s)
For MiNP (a metabolite of di-isononyl phthalate), in linear regression models adjusted for age and BMI, we found that with increasing quartiles of follicular MiNP, there was a significant trend in the decrease in mean AFC (P-trend = 0.023) and a sugge
{"title":"The impact of follicular fluid phthalate metabolites on the ovarian reserve and ovarian function in Indian women undergoing intracytoplasmic sperm injection","authors":"Firuza Rajesh Parikh M.D., Ph.D. , Shonali Uttamchandani B.Sc. , Sujatha Sawkar M.D. , Madhavi Panpalia M.S. , Nandkishor Naik B.Sc. , Prachi Sinkar M.D. , Dhananjaya Kulkarni Ph.D. , Rajesh Parikh M.D.","doi":"10.1016/j.xfss.2023.11.001","DOIUrl":"10.1016/j.xfss.2023.11.001","url":null,"abstract":"<div><h3>Objective</h3><p><span><span>To investigate the adverse effects of phthalate-induced ovarian toxicity on the ovarian reserve<span> and ovarian function<span><span>. To assess whether the accumulation of higher levels of selected phthalate metabolites in the follicular fluid (FF) of Indian women undergoing </span>intracytoplasmic sperm injection (ICSI) was associated with a decline in their </span></span></span>antral follicle<span> count (AFC) and/or serum antimüllerian hormone (AMH) levels, suggesting a negative impact on the ovarian reserve. To evaluate the effects of follicular phthalate metabolites on peak serum estradiol (E</span></span><sub>2</sub>) levels and the total number of oocytes and mature metaphase II (MII) stage oocytes retrieved to assess the impact of phthalate toxicity on ovarian function.</p></div><div><h3>Design</h3><p><span>A subanalysis of an ongoing prospective cohort study was conducted to examine the association between the levels of six phthalate metabolites, namely, mono-n-butyl phthalate (MBP), mono-ethyl phthalate (MEP), mono-isononyl phthalate (MiNP), mono-isodecyl phthalate (MiDP), mono(2-ethyl-5-oxohexyl) phthalate, and mono(2-ethyl-5-hydroxyhexyl) phthalate, in the FF of Indian women undergoing ICSI and their ovarian reserve markers (AFC and serum AMH levels). To investigate the association of these follicular phthalate metabolite levels with the peak E</span><sub>2</sub> levels and the total number of oocytes and number of MII stage oocytes retrieved.</p></div><div><h3>Setting</h3><p>In vitro fertilization center in a referral hospital in India.</p></div><div><h3>Patient(s)</h3><p>A total of 245 consenting Indian women who had undergone oocyte retrieval<span> between April 2017 and mid-March 2020 were included. Each woman contributed one FF sample to the study. This was screened for six phthalate metabolites. The samples were collected before the coronavirus disease 2019 pandemic.</span></p></div><div><h3>Intervention(s)</h3><p><span>Using liquid chromatography-tandem mass spectrometry, the total levels of six phthalate metabolites were quantified in the FF of 245 women. Using linear regression<span> models that were unadjusted and adjusted for maternal age and body mass index (BMI), we evaluated the association between the follicular metabolites in these women and their AFC, serum AMH levels, peak E</span></span><sub>2</sub> levels, total number of oocytes, and MII stage oocytes.</p></div><div><h3>Main Outcome Measure(s)</h3><p>To evaluate the impact of phthalate-induced ovarian toxicity on the ovarian reserve and ovarian function in Indian women undergoing ICSI by studying their accumulated levels in their FF.</p></div><div><h3>Result(s)</h3><p>For MiNP (a metabolite of di-isononyl phthalate), in linear regression models adjusted for age and BMI, we found that with increasing quartiles of follicular MiNP, there was a significant trend in the decrease in mean AFC (<em>P-</em>trend = 0.023) and a sugge","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 2","pages":"Pages 107-120"},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139466967","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 : 2024-02-01DOI: 10.1016/j.xfss.2023.11.004
Maike K. Sachs M.D. , Sofia Makieva Ph.D. , Ana Velasco Gil , Min Xie Ph.D. , Fabian Ille Ph.D. , Vincent Salvadori , Meret Schmidhauser Ph.D. , Mara D. Saenz-de-Juano Ph.D. , Susanne E. Ulbrich Ph.D. , Brigitte Leeners M.D.
Objective
To compare the transcriptome of human cumulus cells (CCs) from oocytes with different outcomes (pregnancy yes/no, live birth [LB] yes/no), to identify noninvasive biomarkers for oocyte selection as well as new therapeutic targets to increase LB rates from assisted reproductive technologies (ART).
Design
Retrospective observational study.
Settings
This study was conducted at a University Hospital in Switzerland.
Patients
Subfertile couples undergoing controlled ovarian superstimulation and intracytoplasmic sperm injection with subsequent unbiopsied embryo transfer below the female age of 43 years.
Intervention(s)
RNA sequencing of CCs from oocytes results in a pregnancy, no pregnancy, LB, or no LB.
Main outcome Measures
Differential gene expression (DEG) between CCs of oocytes results in “no pregnancy” vs. “pregnancy” and “pregnancy only” vs. “live birth.”
Results
Although RNA sequencing did not reveal DEGs when comparing the transcriptomic profiles of the groups “no pregnancy” with “pregnancy,” we identified 139 DEGs by comparing “pregnancy only” with “live birth,” of which 28 belonged to clusters relevant to successful ART outcomes (i.e., CTGF, SERPINE2, PCK1, HHIP, HS3ST, and BIRC5). A functional enrichment analysis revealed that the transcriptome of CCs associated with LB depicts pathways of extracellular matrix, inflammatory cascades leading to ovulation, cell patterning, proliferation, and differentiation, and silencing pathways leading to apoptosis.
Conclusion
We identified a CCs transcriptomic profile associated with LB after embryo transfer that, after further validation, could serve to predict successful ART outcomes. The definition of relevant pathways of CCs related to oocyte competency contributes to a broader understanding of the cumulus oocyte complex and helps identify further therapeutic targets for improving ART success.
{"title":"Transcriptomic signature of luteinized cumulus cells of oocytes developing to live birth after women received intracytoplasmic sperm injection","authors":"Maike K. Sachs M.D. , Sofia Makieva Ph.D. , Ana Velasco Gil , Min Xie Ph.D. , Fabian Ille Ph.D. , Vincent Salvadori , Meret Schmidhauser Ph.D. , Mara D. Saenz-de-Juano Ph.D. , Susanne E. Ulbrich Ph.D. , Brigitte Leeners M.D.","doi":"10.1016/j.xfss.2023.11.004","DOIUrl":"10.1016/j.xfss.2023.11.004","url":null,"abstract":"<div><h3>Objective</h3><p><span>To compare the transcriptome of human </span>cumulus cells<span> (CCs) from oocytes with different outcomes (pregnancy yes/no, live birth<span> [LB] yes/no), to identify noninvasive biomarkers for oocyte selection as well as new therapeutic targets to increase LB rates from assisted reproductive technologies (ART).</span></span></p></div><div><h3>Design</h3><p>Retrospective observational study.</p></div><div><h3>Settings</h3><p>This study was conducted at a University Hospital in Switzerland.</p></div><div><h3>Patients</h3><p>Subfertile couples undergoing controlled ovarian superstimulation and intracytoplasmic sperm injection<span> with subsequent unbiopsied embryo transfer below the female age of 43 years.</span></p></div><div><h3>Intervention(s)</h3><p>RNA sequencing of CCs from oocytes results in a pregnancy, no pregnancy, LB, or no LB.</p></div><div><h3>Main outcome Measures</h3><p>Differential gene expression (DEG) between CCs of oocytes results in “no pregnancy” vs. “pregnancy” and “pregnancy only” vs. “live birth.”</p></div><div><h3>Results</h3><p><span>Although RNA sequencing did not reveal DEGs when comparing the transcriptomic profiles of the groups “no pregnancy” with “pregnancy,” we identified 139 DEGs by comparing “pregnancy only” with “live birth,” of which 28 belonged to clusters relevant to successful ART outcomes (i.e., </span><span><em>CTGF</em></span>, <span><em>SERPINE2</em></span>, <span><em>PCK1</em></span>, <em>HHIP</em>, <em>HS3ST</em>, and <em>BIRC5</em><span><span>). A functional enrichment analysis revealed that the transcriptome of CCs associated with LB depicts pathways of extracellular matrix, inflammatory cascades leading to </span>ovulation<span>, cell patterning, proliferation, and differentiation, and silencing pathways leading to apoptosis.</span></span></p></div><div><h3>Conclusion</h3><p>We identified a CCs transcriptomic profile associated with LB after embryo transfer that, after further validation, could serve to predict successful ART outcomes. The definition of relevant pathways of CCs related to oocyte competency contributes to a broader understanding of the cumulus oocyte complex and helps identify further therapeutic targets for improving ART success.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 24-38"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138464723","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}
Ovary vitrification is a way for the preservation of fertility in women undergoing chemotherapy and for protecting the valuable or the endangered species. However, cryopreservation of complex tissues, which are composed of different cells and materials, encountered various challenges including oxidative stress damage.
Objectives
This study aimed to evaluate some oxidative stress indices in the vitrified bovine ovaries.
Methods
The pieces of the bovine ovarian cortex (1 × 1 × 1 mm3) were vitrified with final concentrations of ethylene glycol (25%) and glycerol (25%) and 0.5 M sucrose and then, after 48 h, were warmed with descending concentrations (0.5, 0.25, and 0.125 M) of sucrose. The ovaries were processed and some biochemical indicators of oxidative stresses were assayed.
Results
Total antioxidant capacity had a 45% decrease after vitrification (P<.0001). This reduction was associated with a 4 times increase in malondialdehyde (P=.0002) and a 53% decrease in superoxide dismutase (P=.0081). The levels of protein carbonyl in vitrified-warmed ovaries were less than in fresh ovaries (P=.0325). Regression analysis showed that the components of oxidative stress indices in vitrified tissues are different from those of fresh tissues.
Conclusion
An extensive alteration was seen in oxidant/antioxidant balance during vitrification.
{"title":"Redox reactions in vitrified-warmed ovary","authors":"Atefe Rahimi D.V.M , Ali Shahriari Ph.D. , Farid Barati Ph.D.","doi":"10.1016/j.xfss.2023.11.002","DOIUrl":"10.1016/j.xfss.2023.11.002","url":null,"abstract":"<div><h3>Backgrounds</h3><p>Ovary vitrification is a way for the preservation of fertility in women undergoing chemotherapy and for protecting the valuable or the endangered species. However, cryopreservation of complex tissues, which are composed of different cells and materials, encountered various challenges including oxidative stress damage.</p></div><div><h3>Objectives</h3><p>This study aimed to evaluate some oxidative stress indices in the vitrified bovine ovaries.</p></div><div><h3>Methods</h3><p>The pieces of the bovine ovarian cortex (1 × 1 × 1 mm<sup>3</sup><span>) were vitrified with final concentrations of ethylene glycol (25%) and glycerol (25%) and 0.5 M sucrose and then, after 48 h, were warmed with descending concentrations (0.5, 0.25, and 0.125 M) of sucrose. The ovaries were processed and some biochemical indicators of oxidative stresses were assayed.</span></p></div><div><h3>Results</h3><p><span>Total antioxidant capacity had a 45% decrease after vitrification (</span><em>P</em><span><.0001). This reduction was associated with a 4 times increase in malondialdehyde (</span><em>P</em>=.0002) and a 53% decrease in superoxide dismutase (<em>P</em>=.0081). The levels of protein carbonyl in vitrified-warmed ovaries were less than in fresh ovaries (<em>P</em>=.0325). Regression analysis showed that the components of oxidative stress indices in vitrified tissues are different from those of fresh tissues.</p></div><div><h3>Conclusion</h3><p>An extensive alteration was seen in oxidant/antioxidant balance during vitrification.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 39-42"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89720930","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 : 2024-02-01DOI: 10.1016/j.xfss.2023.12.005
Rachel C. Nordberg Ph.D. , Renata S. Magalhaes M.D., Ph.D. , Irene Cervelló Ph.D. , J.Koudy Williams D.V.M. , Anthony Atala M.D. , Elizabeth G. Loboa Ph.D.
Objective
To assess the in vivo biomechanical maturation of tissue-engineered neo-uteri that have previously supported live births in a rabbit model.
Design
Nonclinical animal study.
Setting
University-based research laboratory.
Animals
Eighteen adult female rabbits.
Intervention
Biodegradable poly-DL-lactide-co-glycolide-coated polyglycolic acid scaffolds seeded with autologous uterine-derived endometrial and myometrial cells. Nonseeded scaffolds and seeded, tissue-engineered neo-uteri were implanted into one uterine horn of rabbits for 1, 3, or 6 months, excised, and biomechanically assessed in comparison to native uterine tissue.
Main Outcome Measures
Tensile stress-relaxation testing, strain-to-failure testing, and viscoelastic modeling.
Results
By evaluating the biomechanical data with several viscoelastic models, it was revealed that tissue-engineered uteri were more mechanically robust than nonseeded scaffolds. For example, the 10% instantaneous stress of the tissue-engineered neo-uteri was 2.1 times higher than the nonseeded scaffolds at the 1-month time point, 1.6 times higher at the 3-month time point, and 1.5 times higher at the 6-month time point. Additionally, as the duration of implantation increased, the engineered constructs became more mechanically robust (e.g., 10% instantaneous stress of the tissue-engineered neo-uteri increased from 22 kPa at 1 month to 42 kPa at 6 months). Compared with native tissue values, tissue-engineered neo-uteri achieved or surpassed native tissue values by the 6-month time point.
Conclusion
The present study evaluated the mechanical characteristics of novel tissue-engineered neo-uteri that have previously been reported to support live births in the rabbit model. We demonstrate that the biomechanics of these implants closely resemble those of native tissue, giving further credence to their development as a clinical solution to uterine factor infertility.
{"title":"A biomechanical assessment of tissue-engineered polymer neo-uteri after orthotopic implantation","authors":"Rachel C. Nordberg Ph.D. , Renata S. Magalhaes M.D., Ph.D. , Irene Cervelló Ph.D. , J.Koudy Williams D.V.M. , Anthony Atala M.D. , Elizabeth G. Loboa Ph.D.","doi":"10.1016/j.xfss.2023.12.005","DOIUrl":"10.1016/j.xfss.2023.12.005","url":null,"abstract":"<div><h3>Objective</h3><p>To assess the <em>in vivo</em><span> biomechanical maturation of tissue-engineered neo-uteri that have previously supported live births in a rabbit model.</span></p></div><div><h3>Design</h3><p>Nonclinical animal study.</p></div><div><h3>Setting</h3><p>University-based research laboratory.</p></div><div><h3>Animals</h3><p>Eighteen adult female rabbits.</p></div><div><h3>Intervention</h3><p><span>Biodegradable poly-DL-lactide-co-glycolide-coated polyglycolic acid scaffolds seeded with autologous uterine-derived endometrial and myometrial cells. Nonseeded scaffolds and seeded, tissue-engineered neo-uteri were implanted into one </span>uterine horn of rabbits for 1, 3, or 6 months, excised, and biomechanically assessed in comparison to native uterine tissue.</p></div><div><h3>Main Outcome Measures</h3><p>Tensile stress-relaxation testing, strain-to-failure testing, and viscoelastic modeling.</p></div><div><h3>Results</h3><p>By evaluating the biomechanical data with several viscoelastic models, it was revealed that tissue-engineered uteri were more mechanically robust than nonseeded scaffolds. For example, the 10% instantaneous stress of the tissue-engineered neo-uteri was 2.1 times higher than the nonseeded scaffolds at the 1-month time point, 1.6 times higher at the 3-month time point, and 1.5 times higher at the 6-month time point. Additionally, as the duration of implantation increased, the engineered constructs became more mechanically robust (e.g., 10% instantaneous stress of the tissue-engineered neo-uteri increased from 22 kPa at 1 month to 42 kPa at 6 months). Compared with native tissue values, tissue-engineered neo-uteri achieved or surpassed native tissue values by the 6-month time point.</p></div><div><h3>Conclusion</h3><p>The present study evaluated the mechanical characteristics of novel tissue-engineered neo-uteri that have previously been reported to support live births in the rabbit model. We demonstrate that the biomechanics of these implants closely resemble those of native tissue, giving further credence to their development as a clinical solution to uterine factor infertility.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 58-68"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139038270","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 : 2024-02-01DOI: 10.1016/j.xfss.2023.11.003
Zoran Jason Pavlovic M.D. , Angel Hsin-Yu Pai M.D. , Tzu-Ti Hsiao M.S. , Chih-Feng Yen M.D., Ph.D. , Hasan Alhasan M.D. , Asli Ozmen Ph.D. , Erika P. New M.D. M.P.H. , Xiaofang Guo M.D. , Anthony N. Imudia M.D. , Ozlem Guzeloglu-Kayisli Ph.D. , Charles J. Lockwood M.D. , Umit A. Kayisli Ph.D.
Objective
To study the effect of adenomyosis on the localized expression of the GATA binding proteins 2 and 6 (GATA2 and GATA6) zinc-finger transcription factors that are involved in proliferation of hematopoietic and endocrine cell lineages, cell differentiation, and organogenesis, potentially leading to impaired endometrial implantation.
Design
Laboratory based experimental study.
Setting
Academic hospital and laboratory.
Patients
Human endometrial stromal cells (HESCs) of reproductive age patients, 18–45 years of age, with adenomyosis were compared with patients with no pathology and leiomyomatous uteri as controls (n = 4 in each group, respectively). Additionally, midsecretory phase endometrial sections were obtained from patients with adenomyosis and control patients with leiomyoma (n = 8 in each group, respectively).
Interventions
GATA2 and GATA6 immunohistochemistry and H-SCORE were performed on the midsecretory phase endometrial sections from adenomyosis and leiomyoma control patients (n = 8 each, respectively). Control and adenomyosis patient HESC cultures were treated with placebo or 10-8 M estradiol (E2), or decidualization media (EMC) containing 10-8 M E2, 10-7 M medroxyprogesterone acetate, and 5 × 10-5 M cAMP for 6 and 10 days. Additionally, control HESC cultures (n = 4) were transfected with scrambled small interfering RNA (siRNA) (control) or GATA2-specific siRNAs for 6 days while adenomyosis HESC cultures (n = 4) were transfected with human GATA2 expression vectors to silence or induce GATA2 overexpression.
Main Outcome Measures
Immunohistochemistry was performed to obtain GATA2 and GATA6 H-SCORES in adenomyosis vs. control patient endometrial tissue. Expression of GATA2, GATA6, insulin-like growth factor-binding protein 1 (IGFBP1), prolactin (PRL), progesterone receptor (PGR), estrogen receptor 1 (ESR1), leukemia inhibitory factor (LIF), and Interleukin receptor 11 (IL11R) messenger RNA (mRNA) levels were analyzed using by qPCR with normalization to ACTB. Silencing and overexpression experiments also had the corresponding mRNA levels of the above factors analyzed. Western blot analysis was performed on isolated proteins from transfection experiments.
Results
Immunohistochemistry revealed an overall fourfold lower GATA2 and fourfold higher GATA6 H-SCORE level in the endometrial stromal cells of patients with adenomyosis vs. controls. Decidual induction with EMC resulted in significantly lower GATA2, PGR, PRL and IGFBP1 mRNA levels in HESC cultures from patients with adenomyosis patient vs. controls
{"title":"Dysregulated expression of GATA2 and GATA6 transcription factors in adenomyosis: implications for impaired endometrial receptivity","authors":"Zoran Jason Pavlovic M.D. , Angel Hsin-Yu Pai M.D. , Tzu-Ti Hsiao M.S. , Chih-Feng Yen M.D., Ph.D. , Hasan Alhasan M.D. , Asli Ozmen Ph.D. , Erika P. New M.D. M.P.H. , Xiaofang Guo M.D. , Anthony N. Imudia M.D. , Ozlem Guzeloglu-Kayisli Ph.D. , Charles J. Lockwood M.D. , Umit A. Kayisli Ph.D.","doi":"10.1016/j.xfss.2023.11.003","DOIUrl":"10.1016/j.xfss.2023.11.003","url":null,"abstract":"<div><h3>Objective</h3><p><span>To study the effect of adenomyosis<span> on the localized expression of the GATA binding proteins 2 and 6 (GATA2 and GATA6) zinc-finger transcription factors that are involved in proliferation of hematopoietic and endocrine </span></span>cell lineages<span>, cell differentiation, and organogenesis, potentially leading to impaired endometrial implantation.</span></p></div><div><h3>Design</h3><p>Laboratory based experimental study.</p></div><div><h3>Setting</h3><p>Academic hospital and laboratory.</p></div><div><h3>Patients</h3><p><span>Human endometrial stromal cells (HESCs) of reproductive age patients, 18–45 years of age, with adenomyosis were compared with patients with no pathology and leiomyomatous uteri as controls (n = 4 in each group, respectively). Additionally, midsecretory phase endometrial sections were obtained from patients with adenomyosis and control patients with </span>leiomyoma (n = 8 in each group, respectively).</p></div><div><h3>Interventions</h3><p><span>GATA2<span><span> and GATA6 </span>immunohistochemistry and H-SCORE were performed on the midsecretory phase endometrial sections from adenomyosis and leiomyoma control patients (n = 8 each, respectively). Control and adenomyosis patient HESC cultures were treated with placebo or 10</span></span><sup>-8</sup><span> M estradiol (E2), or decidualization media (EMC) containing 10</span><sup>-8</sup> M E2, 10<sup>-7</sup><span> M medroxyprogesterone acetate, and 5 × 10</span><sup>-5</sup><span> M cAMP for 6 and 10 days. Additionally, control HESC cultures (n = 4) were transfected with scrambled small interfering RNA (siRNA) (control) or </span><em>GATA2</em>-specific siRNAs for 6 days while adenomyosis HESC cultures (n = 4) were transfected with human <em>GATA2</em><span> expression vectors to silence or induce </span><em>GATA2</em> overexpression.</p></div><div><h3>Main Outcome Measures</h3><p><span><span>Immunohistochemistry was performed to obtain GATA2 and GATA6 H-SCORES in adenomyosis vs. control patient endometrial tissue. Expression of GATA2, GATA6, insulin-like growth factor-binding protein 1 (IGFBP1), prolactin (PRL), progesterone receptor (PGR), </span>estrogen receptor 1 (ESR1), leukemia inhibitory factor (LIF), and Interleukin receptor 11 (IL11R) messenger RNA (mRNA) levels were analyzed using by qPCR with normalization to </span><em>ACTB</em><span>. Silencing and overexpression experiments also had the corresponding mRNA levels of the above factors analyzed. Western blot analysis was performed on isolated proteins from transfection experiments.</span></p></div><div><h3>Results</h3><p><span><span>Immunohistochemistry revealed an overall fourfold lower GATA2 and fourfold higher GATA6 H-SCORE level in the endometrial stromal cells of patients with adenomyosis vs. controls. Decidual induction with EMC resulted in significantly lower GATA2, PGR, PRL and IGFBP1 mRNA levels in HESC cultures from patients with adenomyosis patient vs. controls","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 92-103"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135764119","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 : 2024-02-01DOI: 10.1016/j.xfss.2023.12.003
Isaac Stirland B.S. , Murilo Racy Soares Ph.D. , Cristiana Libardi Miranda Furtado Ph.D. , Rosana Maria Dos Reis Ph.D. , Kenneth I. Aston Ph.D. , R. Parker Smith , Timothy G. Jenkins Ph.D.
Objective
To determine whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection affects male reproductive health, considering the many potential factors that contribute to declines in male fertility on a semiglobal scale.
Design
In total, 64 human semen samples—32 treatment and 32 control—were laboratory processed and bioinformatically analyzed to assess differences in DNA methylation patterns. Implementing multiple bioinformatic tools, the analyses conducted will elicit between-group differences with respect to epigenetic age, epigenetic instability, semiglobal, and regional methylation, in addition to methylation patterns as a function of time since infection.
Setting
University hospital.
Patients
The study cohort of 64 individuals was drawn from a larger population of 94 volunteer participants recruited at the Human Reproduction Center at the Clinical Hospital of the Ribeirao Preto Medical School—University of São Paulo between June 2021 and January 2022 as well as in accordance with the ethical guidelines established by the Declaration of Helsinki.
Intervention
Exposure to SARS-CoV-2.
Main Outcome Measure(s)
Effects on male reproductive health were reported as differences in DNA methylation measured using an array. Mean β values at key regulatory loci for human spermatocytes were analyzed and compared between groups. Further analysis of β values using epigenetic age, instability, semiglobal, and regional methylation tools provided an analysis with substantial breadth and depth.
Results
In all analyses, there were no differences between groups. Considering these results, it can be inferred that infection with SARS-CoV-2 does not alter the epigenome of human spermatocytes in significant and/or persistent ways. Tangentially, these data also suggest that human male reproductive health is minimally altered by the virus, or that it is altered in a way that is independent of epigenetic programming.
Conclusion
Infection with SARS-CoV-2 has been reportedly associated with alterations in male fertility. This study asserts that such alterations do not have an epigenetic basis but are likely a result of concomitant symptomatology, i.e., fever and inflammation. Across the multiple bioinformatic analyses conducted, the results of this test did not detect any differences in DNA methylation patterns between coronavirus disease 2019 and noncoronavirus disease semen donor groups.
{"title":"An assessment of alterations to human sperm methylation patterns in coronavirus disease 2019 infected and healthy control males","authors":"Isaac Stirland B.S. , Murilo Racy Soares Ph.D. , Cristiana Libardi Miranda Furtado Ph.D. , Rosana Maria Dos Reis Ph.D. , Kenneth I. Aston Ph.D. , R. Parker Smith , Timothy G. Jenkins Ph.D.","doi":"10.1016/j.xfss.2023.12.003","DOIUrl":"10.1016/j.xfss.2023.12.003","url":null,"abstract":"<div><h3>Objective</h3><p><span>To determine whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection affects </span>male reproductive health<span>, considering the many potential factors that contribute to declines in male fertility on a semiglobal scale.</span></p></div><div><h3>Design</h3><p><span><span>In total, 64 human semen samples—32 treatment and 32 control—were laboratory processed and bioinformatically analyzed to assess differences in DNA </span>methylation patterns. Implementing multiple bioinformatic tools, the analyses conducted will elicit between-group differences with respect to </span>epigenetic age, epigenetic instability, semiglobal, and regional methylation, in addition to methylation patterns as a function of time since infection.</p></div><div><h3>Setting</h3><p>University hospital.</p></div><div><h3>Patients</h3><p>The study cohort of 64 individuals was drawn from a larger population of 94 volunteer participants recruited at the Human Reproduction Center at the Clinical Hospital of the Ribeirao Preto Medical School—University of São Paulo between June 2021 and January 2022 as well as in accordance with the ethical guidelines established by the Declaration of Helsinki.</p></div><div><h3>Intervention</h3><p>Exposure to SARS-CoV-2.</p></div><div><h3>Main Outcome Measure(s)</h3><p><span>Effects on male reproductive health were reported as differences in DNA methylation measured using an array. Mean β values at key regulatory loci for human </span>spermatocytes were analyzed and compared between groups. Further analysis of β values using epigenetic age, instability, semiglobal, and regional methylation tools provided an analysis with substantial breadth and depth.</p></div><div><h3>Results</h3><p>In all analyses, there were no differences between groups. Considering these results, it can be inferred that infection with SARS-CoV-2 does not alter the epigenome of human spermatocytes in significant and/or persistent ways. Tangentially, these data also suggest that human male reproductive health is minimally altered by the virus, or that it is altered in a way that is independent of epigenetic programming.</p></div><div><h3>Conclusion</h3><p>Infection with SARS-CoV-2 has been reportedly associated with alterations in male fertility. This study asserts that such alterations do not have an epigenetic basis but are likely a result of concomitant symptomatology<span>, i.e., fever and inflammation. Across the multiple bioinformatic analyses conducted, the results of this test did not detect any differences in DNA methylation patterns between coronavirus disease 2019 and noncoronavirus disease semen donor groups.</span></p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 2-15"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138614077","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 identify cytokines or extracellular matrix components that contribute to adhesion to, and invasion of, the peritoneum, proximal to lesions in the early phase of endometriosis.
Design
Laboratory-based study.
Setting
University Hospital and Laboratory of Animal Science.
Patients and Animals
Five women with ovarian endometrioma, 138 wild-type (WT) C57BL/6N mice, and 48 Tenascin C (Tnc) knockout (TncKO) mice.
Interventions
To establish a murine endometriosis model, 20 pieces of minced uterine tissue fragments from each horn were administered intraperitoneally to syngeneic mice. Three days later, endometriotic lesions and peritoneal tissues were collected. Separately, we transfected human peritoneal mesothelial cells (HMrSV5) or human endometrial stromal cells (hESCs) with Tnc small interfering ribonucleic acid.
Main Outcome Measures
We employed a polymerase chain reaction array to profile gene expression in the murine peritoneum, in both peritoneum distal to lesions and peritoneum surrounding lesions (PSL). The expression of upregulated genes in the PSL was verified in the peritoneal samples by real-time reverse transcription-polymerase chain reaction. TncKO mice were used to investigate the role of Tnc in the development of endometriosis. We evaluated the proliferative activity or inflammatory state of lesions by Ki67 or CD3 immunostaining. Intraperitoneal distribution of macrophages was assessed by fluorescence-activated cell sorting. Using Tnc small interfering ribonucleic acid, we examined the invasive capacity of hESCs in a coculture system with HMrSV5.
Results
Tnc gene expression was significantly higher in PSL than in peritoneum distal to lesions. The weight and number of TncKO lesions in TncKO hosts were lower than those of WT lesions in WT hosts. In contrast, the weight and number of nonattached TncKO lesions in TncKO hosts were higher than those of nonattached WT lesions in WT hosts. We observed decreased Ki67-positive cells or H-scores for CD3, a lower proportion of M1 macrophages, and a higher proportion of M2 macrophages in TncKO lesions in TncKO recipients. Silencing of Tnc expression in hESCs and HMrSV5 diminished the invasivity of hESCs.
Conclusion
Tnc may be a crucial factor in the development of early peritoneal endometriosis.
{"title":"Role of tenascin C in lesion formation in early peritoneal endometriosis","authors":"Maako Moriyama M.D. , Kazuomi Nakamura Ph.D. , Hiroki Nagata M.D. , Ikumi Wada M.D. , Kei Nagira M.D., Ph.D. , Yukihiro Azuma M.D., Ph.D. , Eri Sato M.D., Ph.D. , Tasuku Harada M.D., Ph.D. , Fuminori Taniguchi M.D., Ph.D.","doi":"10.1016/j.xfss.2023.12.004","DOIUrl":"10.1016/j.xfss.2023.12.004","url":null,"abstract":"<div><h3>Objective</h3><p><span>To identify cytokines or extracellular matrix<span> components that contribute to adhesion to, and invasion of, the peritoneum, proximal to lesions in the early phase of </span></span>endometriosis.</p></div><div><h3>Design</h3><p>Laboratory-based study.</p></div><div><h3>Setting</h3><p>University Hospital and Laboratory of Animal Science.</p></div><div><h3>Patients and Animals</h3><p><span>Five women with ovarian endometrioma, 138 wild-type (WT) C57BL/6N mice, and 48 </span><span><em>Tenascin</em><em> C</em></span> (<em>Tnc</em>) knockout (TncKO) mice.</p></div><div><h3>Interventions</h3><p><span><span>To establish a murine endometriosis model, 20 pieces of minced uterine tissue fragments from each horn were administered intraperitoneally to syngeneic mice. Three days later, endometriotic lesions and peritoneal tissues were collected. Separately, we transfected human peritoneal </span>mesothelial cells<span> (HMrSV5) or human endometrial stromal cells (hESCs) with </span></span><em>Tnc</em><span> small interfering ribonucleic acid.</span></p></div><div><h3>Main Outcome Measures</h3><p><span>We employed a polymerase chain reaction<span> array to profile gene expression in the murine peritoneum, in both peritoneum distal to lesions and peritoneum surrounding lesions (PSL). The expression of upregulated genes in the PSL was verified in the peritoneal samples by real-time reverse transcription-polymerase chain reaction. TncKO mice were used to investigate the role of Tnc in the development of endometriosis. We evaluated the proliferative activity or inflammatory state of lesions by Ki67 or CD3 immunostaining. Intraperitoneal distribution of macrophages was assessed by fluorescence-activated cell sorting. Using </span></span><em>Tnc</em> small interfering ribonucleic acid, we examined the invasive capacity of hESCs in a coculture system with HMrSV5.</p></div><div><h3>Results</h3><p><em>Tnc</em> gene expression was significantly higher in PSL than in peritoneum distal to lesions. The weight and number of TncKO lesions in TncKO hosts were lower than those of WT lesions in WT hosts. In contrast, the weight and number of nonattached TncKO lesions in TncKO hosts were higher than those of nonattached WT lesions in WT hosts. We observed decreased Ki67-positive cells or H-scores for CD3, a lower proportion of M1 macrophages, and a higher proportion of M2 macrophages in TncKO lesions in TncKO recipients. Silencing of <em>Tnc</em> expression in hESCs and HMrSV5 diminished the invasivity of hESCs.</p></div><div><h3>Conclusion</h3><p><em>Tnc</em> may be a crucial factor in the development of early peritoneal endometriosis.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 69-79"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138813718","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 : 2024-02-01DOI: 10.1016/j.xfss.2023.10.005
Xi Guo Ph.D. , Yiping Zhong M.Phil. , Yang Liu M.Phil. , Rihan Wu M.Phil. , Ling Huang Ph.D. , Chuan Huang M.Phil. , Minghui Chen Ph.D.
Objective
To investigate the direct effect of growth differentiation factor 9 (GDF9) on androgen production in human theca cells.
Design
Experimental study.
Setting
Tertiary hospital-based research laboratory.
Patient(s)
Women who underwent in vitro fertilization and intracytoplasmic sperm injections at our clinic were included in this study.
Intervention(s)
Primary cultured human theca cells from women undergoing in vitro fertilization and intracytoplasmic sperm injection treatment were treated with GDF9, an activin receptor-like kinase 5 (ALK5) inhibitor, and a SMAD4 agonist.
Main Outcome Measure(s)
The expression of androgen synthesis-related genes StAR, CYP17A1, and LHCGR, levels of androstenedione and testosterone, phosphorylation of SMAD2/3, and the interaction between bone morphogenic protein-activated type II receptor and ALK5 were evaluated using reverse transcription-quantitative polymerase chain reaction, Western blot, enzyme-linked immunosorbent assays, and coimmunoprecipitation assays, respectively.
Result(s)
Growth differentiation factor 9 decreased StAR, CYP17A1, and LHCGR expression levels in human theca cells, which was prevented by treatment with the ALK5 inhibitor, and suppressed production of androgen in human theca cells. Growth differentiation factor 9 increased SMAD2/3 phosphorylation, and the ALK5 inhibitor also suppressed this effect. Bone morphogenic protein-activated type II receptor and ALK5 bound to each other after GDF9 stimulation. The SMAD4 agonist kartogenin also decreased messenger RNA levels of StAR and CYP17A1 and protein levels of StAR in human theca cells.
Conclusion(s)
Growth differentiation factor 9 can activate the bone morphogenic protein-activated type II receptor-ALK5-SMAD2/3 signaling pathway, suppress CYP17A1 expression, and decrease androgen production in human theca cells.
{"title":"Oocyte-derived growth differentiation factor 9 suppresses the expression of CYP17A1 and androgen production in human theca cells","authors":"Xi Guo Ph.D. , Yiping Zhong M.Phil. , Yang Liu M.Phil. , Rihan Wu M.Phil. , Ling Huang Ph.D. , Chuan Huang M.Phil. , Minghui Chen Ph.D.","doi":"10.1016/j.xfss.2023.10.005","DOIUrl":"10.1016/j.xfss.2023.10.005","url":null,"abstract":"<div><h3>Objective</h3><p><span>To investigate the direct effect of growth differentiation factor 9<span> (GDF9) on androgen production in human </span></span>theca cells.</p></div><div><h3>Design</h3><p>Experimental study.</p></div><div><h3>Setting</h3><p>Tertiary hospital-based research laboratory.</p></div><div><h3>Patient(s)</h3><p>Women who underwent in vitro fertilization and intracytoplasmic sperm injections at our clinic were included in this study.</p></div><div><h3>Intervention(s)</h3><p>Primary cultured human theca cells from women undergoing in vitro fertilization and intracytoplasmic sperm injection treatment were treated with GDF9, an activin receptor-like kinase 5 (ALK5) inhibitor, and a SMAD4 agonist.</p></div><div><h3>Main Outcome Measure(s)</h3><p>The expression of androgen synthesis-related genes <em>StAR</em>, <span><em>CYP17A1</em></span>, and <span><em>LHCGR</em></span><span>, levels of androstenedione<span> and testosterone, phosphorylation of SMAD2/3, and the interaction between bone morphogenic protein-activated type II receptor and ALK5 were evaluated using reverse transcription-quantitative polymerase chain reaction, Western blot, enzyme-linked immunosorbent assays, and coimmunoprecipitation assays, respectively.</span></span></p></div><div><h3>Result(s)</h3><p>Growth differentiation factor 9 decreased <em>StAR, CYP17A1</em>, and <em>LHCGR</em> expression levels in human theca cells, which was prevented by treatment with the ALK5 inhibitor, and suppressed production of androgen in human theca cells. Growth differentiation factor 9 increased SMAD2/3 phosphorylation, and the ALK5 inhibitor also suppressed this effect. Bone morphogenic protein-activated type II receptor and ALK5 bound to each other after GDF9 stimulation. The SMAD4 agonist kartogenin also decreased messenger RNA levels of <em>StAR</em> and <em>CYP17A1</em> and protein levels of StAR in human theca cells.</p></div><div><h3>Conclusion(s)</h3><p><span>Growth differentiation factor 9 can activate the bone morphogenic protein-activated type II receptor-ALK5-SMAD2/3 signaling pathway, suppress </span><em>CYP17A1</em> expression, and decrease androgen production in human theca cells.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 16-23"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136152846","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 : 2024-02-01DOI: 10.1016/j.xfss.2023.10.002
Nikica Zaninovic M.S., Ph.D. , Jose T. Sierra Ph.D. , Jonas E. Malmsten D.P.S. , Zev Rosenwaks M.D.
Objective
To evaluate the degree of agreement of embryo ranking between embryologists and eight artificial intelligence (AI) algorithms.
Design
Retrospective study.
Patient(s)
A total of 100 cycles with at least eight embryos were selected from the Weill Cornell Medicine database. For each embryo, the full-length time-lapse (TL) videos, as well as a single embryo image at 120 hours, were given to five embryologists and eight AI algorithms for ranking.
Embryologists had a high degree of agreement in the overall ranking of 100 cycles with an average Kendall’s tau (K-τ) of 0.70, slightly lower than the interembryologist agreement when using a single image or video (average K-τ = 0.78). Overall agreement between embryologists and the AI algorithms was significantly lower (average K-τ = 0.53) and similar to the observed low inter-AI algorithm agreement (average K-τ = 0.47). Notably, two of the eight algorithms had a very low agreement with other ranking methodologies (average K-τ = 0.05) and between each other (K-τ = 0.01). The average agreement in selecting the best-quality embryo (1/8 in 100 cycles with an expected agreement by random chance of 12.5%; confidence interval [CI]95: 6%–19%) was 59.5% among embryologists and 40.3% for six AI algorithms. The incidence of the agreement for the two algorithms with the low overall agreement was 11.7%. Agreement on selecting the same top two embryos/cycle (expected agreement by random chance corresponds to 25.0%; CI95: 17%–32%) was 73.5% among embryologists and 56.0% among AI methods excluding two discordant algorithms, which had an average agreement of 24.4%, the expected range of agreement by random chance. Intraembryologist ranking agreement (single image vs. video) was 71.7% and 77.8% for single and top two embryos, respectively. Analysis of average raw scores indicated that cycles with low diversity of embryo quality generally resulted in a lower overall agreement between the methods (embryologists and AI models).
Conclusion(s)
To our knowledge, this is the first study that evaluates the level of agreement in ranking embryo quality between different AI algorithms and embryologists. The different concordance methods were consistent and indicated that the highest agreement was intraembryologist agreement, followed by interembryologist agreement. In contrast, the agreement between some of the AI algorithms and embryologists was similar to the inter-AI algorithm agreement, which also showed a wide range of pairwise concordance. Specifically, two AI models showed intra- and interagreement at the level expected from random selection.
{"title":"Embryo ranking agreement between embryologists and artificial intelligence algorithms","authors":"Nikica Zaninovic M.S., Ph.D. , Jose T. Sierra Ph.D. , Jonas E. Malmsten D.P.S. , Zev Rosenwaks M.D.","doi":"10.1016/j.xfss.2023.10.002","DOIUrl":"10.1016/j.xfss.2023.10.002","url":null,"abstract":"<div><h3>Objective</h3><p>To evaluate the degree of agreement of embryo ranking between embryologists and eight artificial intelligence (AI) algorithms.</p></div><div><h3>Design</h3><p>Retrospective study.</p></div><div><h3>Patient(s)</h3><p>A total of 100 cycles with at least eight embryos were selected from the Weill Cornell Medicine database. For each embryo, the full-length time-lapse (TL) videos, as well as a single embryo image at 120 hours, were given to five embryologists and eight AI algorithms for ranking.</p></div><div><h3>Intervention(s)</h3><p>None.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Kendall rank correlation coefficient (Kendall’s τ).</p></div><div><h3>Result(s)</h3><p>Embryologists had a high degree of agreement in the overall ranking of 100 cycles with an average Kendall’s tau (K-τ) of 0.70, slightly lower than the interembryologist agreement when using a single image or video (average K-τ = 0.78). Overall agreement between embryologists and the AI algorithms was significantly lower (average K-τ = 0.53) and similar to the observed low inter-AI algorithm agreement (average K-τ = 0.47). Notably, two of the eight algorithms had a very low agreement with other ranking methodologies (average K-τ = 0.05) and between each other (K-τ = 0.01). The average agreement in selecting the best-quality embryo (1/8 in 100 cycles with an expected agreement by random chance of 12.5%; confidence interval [CI]95: 6%–19%) was 59.5% among embryologists and 40.3% for six AI algorithms. The incidence of the agreement for the two algorithms with the low overall agreement was 11.7%. Agreement on selecting the same top two embryos/cycle (expected agreement by random chance corresponds to 25.0%; CI95: 17%–32%) was 73.5% among embryologists and 56.0% among AI methods excluding two discordant algorithms, which had an average agreement of 24.4%, the expected range of agreement by random chance. Intraembryologist ranking agreement (single image vs. video) was 71.7% and 77.8% for single and top two embryos, respectively. Analysis of average raw scores indicated that cycles with low diversity of embryo quality generally resulted in a lower overall agreement between the methods (embryologists and AI models).</p></div><div><h3>Conclusion(s)</h3><p>To our knowledge, this is the first study that evaluates the level of agreement in ranking embryo quality between different AI algorithms and embryologists. The different concordance methods were consistent and indicated that the highest agreement was intraembryologist agreement, followed by interembryologist agreement. In contrast, the agreement between some of the AI algorithms and embryologists was similar to the inter-AI algorithm agreement, which also showed a wide range of pairwise concordance. Specifically, two AI models showed intra- and interagreement at the level expected from random selection.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 50-57"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41222069","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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