Pub Date : 2024-05-01DOI: 10.1016/j.xfss.2024.02.002
Meghan Robinson B.Sc. , Kevin Zhou B.Sc. , Sonia H.Y. Kung M.Sc. , Fatih Karaoğlanoğlu M.Sc. , Andrew Golin M.D. , Armita Safa M.Sc. , Charley Cai B.Sc. , Luke Witherspoon M.D. , Faraz Hach Ph.D. , Ryan Flannigan M.D.
Objective
To determine if early spermatocytes can be enriched from a human testis biopsy using fluorescence-activated cell sorting (FACS).
Design
Potential surface markers for early spermatocytes were identified using bioinformatics analysis of single-cell RNA–sequenced human testis tissue. Testicular sperm extraction samples from three participants with normal spermatogenesis were digested into single-cell suspensions and cryopreserved. Two to four million cells were obtained from each and sorted by FACS as separate biologic replicates using antibodies for the identified surface markers. A portion from each biopsy remained unsorted to serve as controls. The sorted cells were then characterized for enrichment of early spermatocytes.
Setting
A laboratory study.
Patients
Three men with a diagnosis of obstructive azoospermia (age range, 30–40 years).
Intervention
None.
Main Outcome Measures
Sorted cells were characterized for RNA expression of markers encompassing the stages of spermatogenesis. Sorting markers were validated by their reactivity on human testis formalin-fixed paraffin-embedded tissue.
Results
Serine protease 50 (TSP50) and SWI5-dependent homologous recombination repair protein 1 were identified as potential surface proteins specific for early spermatocytes. After FACS sorting, the TSP50-sorted populations accounted for 1.6%–8.9% of total populations and exhibited the greatest average-fold increases in RNA expression for the premeiotic marker stimulated by retinoic acid (STRA8), by 23-fold. Immunohistochemistry showed the staining pattern for TSP50 to be strong in premeiotic undifferentiated embryonic cell transcription factor 1−/doublesex and Mab-3 related transcription factor 1−/STRA8+ spermatogonia as well as SYCP3+/protamine 2− spermatocytes.
Conclusion
This work shows that TSP50 can be used to enrich early STRA8-expressing spermatocytes from human testicular biopsies, providing a means for targeted single-cell RNA sequencing analysis and in vitro functional interrogation of germ cells during the onset of meiosis. This could enable investigation into details of the regulatory pathways underlying this critical stage of spermatogenesis, previously difficult to enrich from whole tissue samples.
{"title":"A novel sorting method for the enrichment of early human spermatocytes from clinical biopsies","authors":"Meghan Robinson B.Sc. , Kevin Zhou B.Sc. , Sonia H.Y. Kung M.Sc. , Fatih Karaoğlanoğlu M.Sc. , Andrew Golin M.D. , Armita Safa M.Sc. , Charley Cai B.Sc. , Luke Witherspoon M.D. , Faraz Hach Ph.D. , Ryan Flannigan M.D.","doi":"10.1016/j.xfss.2024.02.002","DOIUrl":"10.1016/j.xfss.2024.02.002","url":null,"abstract":"<div><h3>Objective</h3><p>To determine if early spermatocytes can be enriched from a human testis biopsy using fluorescence-activated cell sorting (FACS).</p></div><div><h3>Design</h3><p>Potential surface markers for early spermatocytes were identified using bioinformatics analysis of single-cell RNA–sequenced human testis tissue. Testicular sperm extraction samples from three participants with normal spermatogenesis were digested into single-cell suspensions and cryopreserved. Two to four million cells were obtained from each and sorted by FACS as separate biologic replicates using antibodies for the identified surface markers. A portion from each biopsy remained unsorted to serve as controls. The sorted cells were then characterized for enrichment of early spermatocytes.</p></div><div><h3>Setting</h3><p>A laboratory study.</p></div><div><h3>Patients</h3><p>Three men with a diagnosis of obstructive azoospermia (age range, 30–40 years).</p></div><div><h3>Intervention</h3><p>None.</p></div><div><h3>Main Outcome Measures</h3><p>Sorted cells were characterized for RNA expression of markers encompassing the stages of spermatogenesis. Sorting markers were validated by their reactivity on human testis formalin-fixed paraffin-embedded tissue.</p></div><div><h3>Results</h3><p>Serine protease 50 (TSP50) and SWI5-dependent homologous recombination repair protein 1 were identified as potential surface proteins specific for early spermatocytes. After FACS sorting, the TSP50-sorted populations accounted for 1.6%–8.9% of total populations and exhibited the greatest average-fold increases in RNA expression for the premeiotic marker stimulated by retinoic acid (STRA8), by 23-fold. Immunohistochemistry showed the staining pattern for TSP50 to be strong in premeiotic undifferentiated embryonic cell transcription factor 1<sup>−</sup>/doublesex and Mab-3 related transcription factor 1<sup>−</sup>/STRA8<sup>+</sup> spermatogonia as well as SYCP3<sup>+</sup>/protamine 2<sup>−</sup> spermatocytes.</p></div><div><h3>Conclusion</h3><p>This work shows that TSP50 can be used to enrich early STRA8-expressing spermatocytes from human testicular biopsies, providing a means for targeted single-cell RNA sequencing analysis and in vitro functional interrogation of germ cells during the onset of meiosis. This could enable investigation into details of the regulatory pathways underlying this critical stage of spermatogenesis, previously difficult to enrich from whole tissue samples.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 2","pages":"Pages 130-140"},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139900996","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 : 2024-05-01DOI: 10.1016/j.xfss.2023.12.002
Samantha B. Schon M.D. , Lindsay Moritz Ph.D. , Mashiat Rabbani B.S. , Julia Meguid , Brock R. Juliano Ph.D. , Brandon T. Ruotolo Ph.D. , Kenneth Aston Ph.D. , Saher Sue Hammoud Ph.D.
Objective
To perform a comprehensive assessment of protamine (P) isoforms and modifications in human sperm with the aim of identifying how P modifications and isoforms are altered in men with reduced sperm motility and low sperm count.
Design
Cross-sectional.
Setting
Academic medical center.
Patients
A total of 18 men with prior reported pregnancy and normozoospermia (normal sperm), 14 men from couples with infertility and asthenozoospermia (reduced sperm motility), and 24 men from couples with infertility and oligoasthenoteratozoospermia (low sperm count and motility and abnormal sperm morphology).
Intervention(s)
Not applicable.
Main Outcome Measure(s)
Proteomic assessment using both top-down and bottom-up liquid chromatography mass spectrometry (MS) analysis.
Results
A total of 13 posttranslational modifications were identified on P1 and P2 using bottom-up MS, including both phosphorylation and methylation. Top-down MS revealed an unmodified and phosphorylated isoform of P1 and the 3 major isoforms of P2, HP2, HP3, and HP4. Protamine 1 phosphorylation was overall higher in men with male factor infertility compared with those with normal semen analysis (40.5% vs. 32.6). There was no difference in P posttranslational modifications or isoforms of P2 in men with normal vs. abnormal fertility.
Conclusion
Human protamines bear a number of posttranslational modifications, with alterations in P1 phosphorylation noted in the setting of male factor infertility.
{"title":"Proteomic analysis of human sperm reveals changes in protamine 1 phosphorylation in men with infertility","authors":"Samantha B. Schon M.D. , Lindsay Moritz Ph.D. , Mashiat Rabbani B.S. , Julia Meguid , Brock R. Juliano Ph.D. , Brandon T. Ruotolo Ph.D. , Kenneth Aston Ph.D. , Saher Sue Hammoud Ph.D.","doi":"10.1016/j.xfss.2023.12.002","DOIUrl":"10.1016/j.xfss.2023.12.002","url":null,"abstract":"<div><h3>Objective</h3><p><span>To perform a comprehensive assessment of protamine (P) isoforms and modifications in human sperm with the aim of identifying how P modifications and isoforms are altered in men with reduced </span>sperm motility and low sperm count.</p></div><div><h3>Design</h3><p>Cross-sectional.</p></div><div><h3>Setting</h3><p>Academic medical center.</p></div><div><h3>Patients</h3><p>A total of 18 men with prior reported pregnancy and normozoospermia (normal sperm), 14 men from couples with infertility and asthenozoospermia (reduced sperm motility), and 24 men from couples with infertility and oligoasthenoteratozoospermia (low sperm count and motility and abnormal sperm morphology).</p></div><div><h3>Intervention(s)</h3><p>Not applicable.</p></div><div><h3>Main Outcome Measure(s)</h3><p><span>Proteomic assessment using both top-down and bottom-up </span>liquid chromatography mass spectrometry (MS) analysis.</p></div><div><h3>Results</h3><p><span>A total of 13 posttranslational modifications were identified on P1 and P2 using bottom-up MS, including both phosphorylation and </span>methylation<span>. Top-down MS revealed an unmodified and phosphorylated isoform of P1 and the 3 major isoforms of P2, HP2, HP3, and HP4. Protamine 1 phosphorylation was overall higher in men with male factor infertility compared with those with normal semen analysis (40.5% vs. 32.6). There was no difference in P posttranslational modifications or isoforms of P2 in men with normal vs. abnormal fertility.</span></p></div><div><h3>Conclusion</h3><p>Human protamines bear a number of posttranslational modifications, with alterations in P1 phosphorylation noted in the setting of male factor infertility.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 2","pages":"Pages 121-129"},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138607666","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 : 2024-05-01DOI: 10.1016/j.xfss.2024.03.003
Gizem Nur Koyan Karadeniz M.D. , Ozan Karadeniz M.D. , Eralp Bulutlar M.D. , Bugra Yilmaz M.D. , Asuman Gedikbasi M.D. , Hilal Serap Arslan M.D. , Berna Aslan Cetin M.D. , İbrahim Polat M.D.
Objective
To compare salpingectomy and detorsion procedures and investigate the biochemical and histopathological changes in the fallopian tubes in the experimentally isolated fallopian tube torsion model in rats.
Design
Experimental study.
Setting
Experimental surgery laboratory in a training and research hospital.
Animal(s)
Twenty-seven Sprague-Dawley rats in the reproductive period.
Intervention(s)
Group 1, control group (n = 6); group 2, bilateral total salpingectomy group after 4 hours of tubal ischemia (n = 7); group 3: 4 hours of bilateral tubal ischemia plus 1 week of reperfusion (n = 7); and group 4, 4-hour period of bilateral tubal ischemia plus 30 days of reperfusion (n = 7). A 22-gauge catheter was administered before and after surgery using methylene blue through the uterine horn of the rat to evaluate tubal patency.
Main Outcome Measure(s)
Preoperative and postoperative serum antimüllerian hormone (AMH) levels, histopathological examination of the rat tuba uterine and histopathological damage scores, antioxidant compounds (superoxide dismutase [SOD], catalase, and glutathione peroxidase [GSH-Px]), and oxidative stress end product levels (malondialdehyde [MDA] and 8-hydroxy-2′-deoxyguanosine [8-OHdG]).
Result(s)
Although a significant difference was observed in the tissue SOD, GSH-Px, MDA, and 8-OHdG values, no significant difference was observed between the groups in serum samples. The tissue SOD and tissue GSH-Px levels in group 2 significantly decreased, and a significant increase was observed in the tissue MDA and 8-OHdG values in group 2. Among the histopathological parameters, epithelial changes, vascular congestion, and the total fallopian tube mean damage score of 4 showed a significant decrease in group 4. When the methylene blue transitions before and after ischemia-reperfusion injury were compared, the values of the methylene blue transition after ischemia-reperfusion injury in groups 2–4 significantly decreased. When the serum AMH levels were analyzed, the postoperative AMH value in group 2 significantly increased.
Conclusion(s)
This study reveals that biochemical and histopathological improvement is observed in the fallopian tube tissues gradually when the detorsion procedure is performed for the necrotized tubal tissue instead of salpingectomy. Although there is restoration of epithelial integrity after reperfusion, tubal passage remains absent.
Clinical Trial Registration Number
This study was approved by the Local Ethics Committee for Animal Experiments of the Health Sciences University, Istanbul Hamidiye Medicine Faculty (approval number 27.05.2022-9269). The study followed the ethics standards recommended by the Declaration of Helsinki.
{"title":"Comparison of salpingectomy and tubal detorsion procedures after experimental ischemia-reperfusion injury in a rat fallopian tube model: biochemical and histopathological evaluation","authors":"Gizem Nur Koyan Karadeniz M.D. , Ozan Karadeniz M.D. , Eralp Bulutlar M.D. , Bugra Yilmaz M.D. , Asuman Gedikbasi M.D. , Hilal Serap Arslan M.D. , Berna Aslan Cetin M.D. , İbrahim Polat M.D.","doi":"10.1016/j.xfss.2024.03.003","DOIUrl":"10.1016/j.xfss.2024.03.003","url":null,"abstract":"<div><h3>Objective</h3><p>To compare salpingectomy and detorsion procedures and investigate the biochemical and histopathological changes in the fallopian tubes in the experimentally isolated fallopian tube torsion model in rats.</p></div><div><h3>Design</h3><p>Experimental study.</p></div><div><h3>Setting</h3><p>Experimental surgery laboratory in a training and research hospital.</p></div><div><h3>Animal(s)</h3><p>Twenty-seven Sprague-Dawley rats in the reproductive period.</p></div><div><h3>Intervention(s)</h3><p>Group 1, control group (n = 6); group 2, bilateral total salpingectomy group after 4 hours of tubal ischemia (n = 7); group 3: 4 hours of bilateral tubal ischemia plus 1 week of reperfusion (n = 7); and group 4, 4-hour period of bilateral tubal ischemia plus 30 days of reperfusion (n = 7). A 22-gauge catheter was administered before and after surgery using methylene blue through the uterine horn of the rat to evaluate tubal patency.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Preoperative and postoperative serum antimüllerian hormone (AMH) levels, histopathological examination of the rat tuba uterine and histopathological damage scores, antioxidant compounds (superoxide dismutase [SOD], catalase, and glutathione peroxidase [GSH-Px]), and oxidative stress end product levels (malondialdehyde [MDA] and 8-hydroxy-2′-deoxyguanosine [8-OHdG]).</p></div><div><h3>Result(s)</h3><p>Although a significant difference was observed in the tissue SOD, GSH-Px, MDA, and 8-OHdG values, no significant difference was observed between the groups in serum samples. The tissue SOD and tissue GSH-Px levels in group 2 significantly decreased, and a significant increase was observed in the tissue MDA and 8-OHdG values in group 2. Among the histopathological parameters, epithelial changes, vascular congestion, and the total fallopian tube mean damage score of 4 showed a significant decrease in group 4. When the methylene blue transitions before and after ischemia-reperfusion injury were compared, the values of the methylene blue transition after ischemia-reperfusion injury in groups 2–4 significantly decreased. When the serum AMH levels were analyzed, the postoperative AMH value in group 2 significantly increased.</p></div><div><h3>Conclusion(s)</h3><p>This study reveals that biochemical and histopathological improvement is observed in the fallopian tube tissues gradually when the detorsion procedure is performed for the necrotized tubal tissue instead of salpingectomy. Although there is restoration of epithelial integrity after reperfusion, tubal passage remains absent.</p></div><div><h3>Clinical Trial Registration Number</h3><p>This study was approved by the Local Ethics Committee for Animal Experiments of the Health Sciences University, Istanbul Hamidiye Medicine Faculty (approval number 27.05.2022-9269). The study followed the ethics standards recommended by the Declaration of Helsinki.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 2","pages":"Pages 195-203"},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140769109","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 : 2024-05-01DOI: 10.1016/j.xfss.2023.10.003
Julienne Chaqour B.S. , Meghan C.H. Ozcan M.D. , Payton De La Cruz M.S. , Morgan F. Woodman-Sousa B.S. , Julia N. McAdams B.S. , Kathryn J. Grive Ph.D.
Objective
To investigate the long-term effects of in utero taxane exposure on exposed daughters’ ovarian reserve and reproductive potential.
Design
Pregnant dams were treated with a single, human-relevant animal-equivalent dose of saline, docetaxel, or paclitaxel at embryonic day 16.5. In utero-exposed daughters were aged to multiple postnatal time points for ovarian and endocrine analysis or were bred to assess fertility and fecundity. Granddaughters of treated dams were assessed also for ovarian follicle composition and atresia.
Setting
Laboratory study.
Animals
C57BL/6 mice.
Intervention(s)
In utero exposure to saline, docetaxel, or paclitaxel.
Main Outcome Measure(s)
Ovarian follicle composition, rates of follicle atresia, and rates of multioocyte follicles were analyzed in all exposure groups. Serum hormone levels and oocyte retrieval outcomes following ovarian hyperstimulation were also assessed. Finally, animals from all exposure groups were bred with the number of litters, pups per litter, live births, interlitter time interval, and age at the last litter analyzed.
Result(s)
We found that docetaxel and paclitaxel exposure in utero results in ovarian toxicity later in life, significantly affecting folliculogenesis as well as increasing the rate of follicular abnormalities, including follicle atresia and multioocyte follicles. Furthermore, viability staining indicates that the ovaries of daughters exposed to taxanes in utero demonstrate a significantly higher number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive follicles. Hormone measurements also revealed that serum follicle-stimulating hormone concentration was significantly altered in taxane-exposed daughters, with the ratio of luteinizing hormone to follicle-stimulating hormone significantly elevated, specifically after paclitaxel exposure, coincident with the inability of these animals to properly respond to ovarian stimulation. Breeding studies over the course of a year also suggest that these taxane-exposed mice are fertile, although the duration of their fertility is shortened and they produce significantly fewer litters. Finally, ovarian effects are apparent in granddaughters of mice treated with docetaxel, suggesting persistent and multigenerational effects of taxane exposure.
Conclusion(s)
Our studies demonstrate that in utero exposure to taxane-based therapy during late gestation has a significant effect on the long-term reproductive health of exposed daughters (as well as their daughters) and will be instrumental in helping clinicians better understand which chemotherapies for maternal malignancy are least detrimental to a developing fetus.
{"title":"Effects of maternal taxane chemotherapy exposure on daughters’ ovarian reserve and fertility potential","authors":"Julienne Chaqour B.S. , Meghan C.H. Ozcan M.D. , Payton De La Cruz M.S. , Morgan F. Woodman-Sousa B.S. , Julia N. McAdams B.S. , Kathryn J. Grive Ph.D.","doi":"10.1016/j.xfss.2023.10.003","DOIUrl":"10.1016/j.xfss.2023.10.003","url":null,"abstract":"<div><h3>Objective</h3><p>To investigate the long-term effects of in utero taxane exposure on exposed daughters’ ovarian reserve and reproductive potential.</p></div><div><h3>Design</h3><p>Pregnant dams were treated with a single, human-relevant animal-equivalent dose of saline, docetaxel, or paclitaxel at embryonic day 16.5. In utero-exposed daughters were aged to multiple postnatal time points for ovarian and endocrine analysis or were bred to assess fertility and fecundity. Granddaughters of treated dams were assessed also for ovarian follicle composition and atresia.</p></div><div><h3>Setting</h3><p>Laboratory study.</p></div><div><h3>Animals</h3><p>C57BL/6 mice.</p></div><div><h3>Intervention(s)</h3><p>In utero exposure to saline, docetaxel, or paclitaxel.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Ovarian follicle composition, rates of follicle atresia, and rates of multioocyte follicles were analyzed in all exposure groups. Serum hormone levels and oocyte retrieval outcomes following ovarian hyperstimulation were also assessed. Finally, animals from all exposure groups were bred with the number of litters, pups per litter, live births, interlitter time interval, and age at the last litter analyzed.</p></div><div><h3>Result(s)</h3><p>We found that docetaxel and paclitaxel exposure in utero results in ovarian toxicity later in life, significantly affecting folliculogenesis as well as increasing the rate of follicular abnormalities, including follicle atresia and multioocyte follicles. Furthermore, viability staining indicates that the ovaries of daughters exposed to taxanes in utero demonstrate a significantly higher number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive follicles. Hormone measurements also revealed that serum follicle-stimulating hormone concentration was significantly altered in taxane-exposed daughters, with the ratio of luteinizing hormone to follicle-stimulating hormone significantly elevated, specifically after paclitaxel exposure, coincident with the inability of these animals to properly respond to ovarian stimulation. Breeding studies over the course of a year also suggest that these taxane-exposed mice are fertile, although the duration of their fertility is shortened and they produce significantly fewer litters. Finally, ovarian effects are apparent in granddaughters of mice treated with docetaxel, suggesting persistent and multigenerational effects of taxane exposure.</p></div><div><h3>Conclusion(s)</h3><p>Our studies demonstrate that in utero exposure to taxane-based therapy during late gestation has a significant effect on the long-term reproductive health of exposed daughters (as well as their daughters) and will be instrumental in helping clinicians better understand which chemotherapies for maternal malignancy are least detrimental to a developing fetus.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 2","pages":"Pages 141-153"},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2666335X23000599/pdfft?md5=9eb1c3d8c0989508e651c49d1b3d4dce&pid=1-s2.0-S2666335X23000599-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136152202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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}
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