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Challenges in Diagnosis and Treatment of Infertility
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Challenges in Diagnosis and Treatment of Infertility

A special issue of Journal of Clinical Medicine (ISSN 2077-0383). This special issue belongs to the section "Reproductive Medicine & Andrology".

Deadline for manuscript submissions: closed (25 September 2024) | Viewed by 16435

Special Issue Editor

Prof. Dr. Konstantinos A. Zikopoulos

E-Mail Website
Guest Editor
Assisted Reproduction Unit, Department of Obstetrics and Gynaecology, Faculty of Medicine, University of Ioannina, 45500 Ioannina, Greece
Interests: research and practice in gynecology endocrinology and medical assisted reproduction

Special Issue Information

Dear Colleagues,

The diagnosis and treatment of infertility are challenging due to issues related to precision required to address individual medical needs and personalized assisted reproduction applications. Age-related or gamete-depletion-related subfertility raises concerns about best practice modalities to warrant early diagnoses and prompt treatments to avoid further fertility loss or a significant reduction in patients’ chances to conceive. Lately, technical advances in preimplantation diagnostic approaches, especially via the introduction of NGS, have offered novel insights into the genetic makeup of preimplantation embryos and even the clinical exploitation of mosaic embryos. Additionally, the treatment outcomes of cell therapies and growth factors showed significant improvements in the regeneration of ovaries and the uterus following the advent of PRP therapy. Furthermore, in all cases, technological innovations such as minimally invasive or non-invasive preimplantation genetic diagnosis improved the prognosis of ART and the prospects for a wider application. It is thus important to closely monitor progress in the evaluation and treatment protocol for fertility. This Special Issue of the Journal of Clinical Medicine focuses on the recent clinical advances in fertility preservation diagnosis and treatment, as well as the improvement of prospects for the conception of and reduction in the time to pregnancy.

Prof. Dr. Konstantinos A. Zikopoulos
Guest Editor

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Keywords

  • fertility
  • regeneration
  • age-related infertility
  • preimplantation diagnosis
  • precision assessment
  • individualized fertility needs
  • clinical diagnosis
  • in vitro fertilization

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Related Special Issue

Published Papers (9 papers)

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Research

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14 pages, 1295 KiB  
Article
Association of Obesity with Telomere Length in Human Sperm
by Efthalia Moustakli, Athanasios Zikopoulos, Charikleia Skentou, Stefanos Dafopoulos, Sofoklis Stavros, Konstantinos Dafopoulos, Peter Drakakis, Ioannis Georgiou and Athanasios Zachariou
J. Clin. Med. 2024, 13(7), 2150; https://doi.org/10.3390/jcm13072150 - 8 Apr 2024
Cited by 1 | Viewed by 2128
Abstract
Background: Telomere attrition and mitochondrial dysfunction are two fundamental aspects of aging. Calorie restriction (CR) is the best strategy to postpone aging since it can enhance telomere attrition, boost antioxidant capacity, and lower the generation of reactive oxygen species (ROS). Since ROS is [...] Read more.
Background: Telomere attrition and mitochondrial dysfunction are two fundamental aspects of aging. Calorie restriction (CR) is the best strategy to postpone aging since it can enhance telomere attrition, boost antioxidant capacity, and lower the generation of reactive oxygen species (ROS). Since ROS is produced by mitochondria and can readily travel to cell nuclei, it is thought to be a crucial molecule for information transfer between mitochondria and cell nuclei. Important variables that affect the quality and functionality of sperm and may affect male reproductive health and fertility include telomere length, mitochondrial content, and the ratio of mitochondrial DNA (mtDNA) to nuclear DNA (nDNA). Telomere damage results from mitochondrial failure, whereas nuclear DNA remains unaffected. This research aims to investigate potential associations between these three variables and how they might relate to body mass index. Methods: Data were collected from 82 men who underwent IVF/ICSI at the University Hospital of Ioannina’s IVF Unit in the Obstetrics and Gynecology Department. Evaluations included sperm morphology, sperm count, sperm motility, and participant history. To address this, male participants who were categorized into three body mass index (ΒΜΙ) groups—normal, overweight, and obese—had their sperm samples tested. Results: For both the normal and overweight groups, our results show a negative connection between relative telomere length and ΒΜI. As an illustration of a potential connection between mitochondrial health and telomere maintenance, a positive correlation was found for the obese group. Only the obese group’s results were statistically significant (p < 0.05). More evidence that longer telomeres are associated with lower mitochondrial content can be found in the negative connection between telomere length and mitochondrial content in both the normal and overweight groups. However, the obese group showed a positive association. The data did not reach statistical significance for any of the three groups. These associations may affect sperm quality since telomere length and mitochondrial concentration are indicators of cellular integrity and health. Moreover, the ratio of mtDNA to nDNA was positively correlated with the relative telomere lengths of the obese group, but negatively correlated with the normal and overweight groups. In every group that was studied, the results were not statistically significant. According to this, male fertility may be negatively impacted by an imbalance in the copy number of the mitochondrial genome compared to the nuclear DNA in sperm. Conclusions: Essentially, the goal of our work is to determine whether mitochondria and telomere length in human sperm interact. Understanding these connections may aid in the explanation of some male infertility causes and possibly contribute to the creation of new treatment modalities for problems pertaining to reproductive health. The functional implications of these connections and their applications in therapeutic settings require further investigation. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)
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10 pages, 977 KiB  
Article
ANRIL rs4977574 Gene Polymorphism in Women with Recurrent Pregnancy Loss
by Panagiotis Cherouveim, Despoina Mavrogianni, Eirini Drakaki, Anastasios Potiris, Athanasios Zikopoulos, Myrto Papamentzelopoulou, Konstantina Kouvoutsaki, Nikolaos Machairiotis, Theodoros Karampitsakos, Chara Skentou, Ekaterini Domali, Nikolaos Vrachnis, Peter Drakakis and Sofoklis Stavros
J. Clin. Med. 2023, 12(18), 5944; https://doi.org/10.3390/jcm12185944 - 13 Sep 2023
Cited by 2 | Viewed by 1087
Abstract
Background: ANRIL rs4977574 gene polymorphism has been associated with arterial thrombosis and cardiovascular disease development. ANRIL rs4977574 gene polymorphism could also be associated with recurrent pregnancy loss (RPL) since there is increasing evidence in favor of a potential shared pathophysiological mechanism with cardiovascular [...] Read more.
Background: ANRIL rs4977574 gene polymorphism has been associated with arterial thrombosis and cardiovascular disease development. ANRIL rs4977574 gene polymorphism could also be associated with recurrent pregnancy loss (RPL) since there is increasing evidence in favor of a potential shared pathophysiological mechanism with cardiovascular disease, potentially through arterial thrombosis. This study’s goal is to investigate the differences in ANRIL rs4977574 gene polymorphism between women with and without RPL, if any, as well as a potential association with the number of pregnancy losses. Methods: DNA was isolated from peripheral blood samples, and the sequence containing the polymorphism of interest was amplified with PCR. Results were visualized under UV light following electrophoresis in 3% agarose gel with ethidium bromide. ANRIL rs4977574 (A>G) prevalence was compared between 56 women with and 69 without RPL. Results were adjusted for women’s age and BMI, while a stratified analysis was performed according to number of pregnancy losses. Results: Allele A was significantly more prevalent in the control group compared to RPL women [31 (44.9%) vs. 14 (25%), p = 0.021]. Although not reaching statistical significance, a gradually decreasing prevalence of allele A with an increasing number of pregnancy losses was observed [31 (44.9%) in control, eight (30.7%) with two, six (23.1%) with three, and 0 (0.0%) with four pregnancy losses, p = 0.078]. Results were also similar following adjustment. Conclusions: This is the first study that demonstrates an association between RPL presence and ANRIL rs4977574 gene polymorphism (lower prevalence of allele A), while a difference according to the number of pregnancy losses cannot be excluded. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)
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9 pages, 244 KiB  
Article
Detection of zeb1 Gene in Granulosa Cells in Women Undergoing IVF Treatment
by Ioannis Chrysanthopoulos, Despoina Mavrogianni, Eirini Drakaki, Anastasios Potiris, Athanasios Zikopoulos, Athanasios Zachariou, Ekaterini Domali, Peter Drakakis and Sofoklis Stavros
J. Clin. Med. 2023, 12(17), 5652; https://doi.org/10.3390/jcm12175652 - 30 Aug 2023
Cited by 1 | Viewed by 1266
Abstract
Background: ZEB1 plays a role in epithelial-to-mesenchymal transition and acts as a repressor of E-cadherin, TGF-β, and Wnt/β-catenin. Since ZEB1 protein is expressed in estrogen-responsive tissues, and expression of the gene in the normal ovary and endometrium is positively correlated with high estrogen [...] Read more.
Background: ZEB1 plays a role in epithelial-to-mesenchymal transition and acts as a repressor of E-cadherin, TGF-β, and Wnt/β-catenin. Since ZEB1 protein is expressed in estrogen-responsive tissues, and expression of the gene in the normal ovary and endometrium is positively correlated with high estrogen levels, we performed a direct analysis of granulosa cell samples to determine whether there are any significant changes in zeb1 expression during folliculogenesis. Methods: ZEB1 expression levels were measured in the granulosa cells of 56 infertile women undergoing IVF treatment. RNA extraction from granulosa cells was performed along with reverse transcription quantitative polymerase chain reaction (RT-qPCR) with SYBR Green I to determine zeb1 gene expression levels. Statistical analysis was performed by using t-test, while possible correlations of the expression of ZEB1 protein with body mass index (BMI), age, number of oocytes, and oocyte maturation were investigated. Results: Zeb1 gene expression levels correlate significantly with body mass index (BMI) and age, but not with oocyte number and oocyte maturation stage. Obese women demonstrate a higher expression level of zeb1 gene compared to normal and overweight women. Moreover, zeb1 gene is overexpressed in women aged 35–40 years old and is under-expressed in women >40 years old. Conclusions: ZEB1 expression should be further investigated as it may unveil new potential findings of the zeb1 gene’s role in female fertility and its use as a biomarker in fertility workups. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)

Review

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19 pages, 363 KiB  
Review
The Roles of GDF-9, BMP-15, BMP-4 and EMMPRIN in Folliculogenesis and In Vitro Fertilization
by Serafeim Fountas, Efthymia Petinaki, Stamatis Bolaris, Magdalini Kargakou, Stefanos Dafopoulos, Athanasios Zikopoulos, Efthalia Moustakli, Sotirios Sotiriou and Konstantinos Dafopoulos
J. Clin. Med. 2024, 13(13), 3775; https://doi.org/10.3390/jcm13133775 - 27 Jun 2024
Viewed by 1365
Abstract
Growth differentiation factor 9 (GDF-9) contributes to early ovarian development and oocyte survival. Higher concentrations of GDF-9 in follicular fluid (FF) are associated with oocyte nuclear maturation and optimal embryo development. In in vitro fertilization (IVF), GDF-9 affects the ability of the oocyte [...] Read more.
Growth differentiation factor 9 (GDF-9) contributes to early ovarian development and oocyte survival. Higher concentrations of GDF-9 in follicular fluid (FF) are associated with oocyte nuclear maturation and optimal embryo development. In in vitro fertilization (IVF), GDF-9 affects the ability of the oocyte to fertilize and subsequent embryonic development. Bone morphogenetic protein 15 (BMP-15) is involved in the regulation of ovarian function and affects oocyte development. During IVF, BMP-15 contributes to the formation of competent blastocysts. BMP-15 may play a role in embryo implantation by affecting endometrial receptivity. Bone morphogenetic protein 4 (BMP-4) is involved in the regulation of follicle growth and development and affects granulosa cell (GC) differentiation. In relation to IVF, BMP-4 is important for embryonic development, influences cell fate and differentiation, and plays a role in facilitating embryo–endometrial interactions during the implantation process. Extracellular matrix metalloproteinase inducer (EMMPRIN) is associated with ovulation and follicle rupture, promotes the release of mature eggs, and affects the modification of the extracellular matrix of the follicular environment. In IVF, EMMPRIN is involved in embryo implantation by modulating the adhesive properties of endometrial cells and promotes trophoblastic invasion, which is essential for pregnancy to occur. The purpose of the current article is to review the studies and recent findings of GDF-9, BMP-15, BMP-4 and EMMPRIN as fundamental factors in normal follicular development and in vitro fertilization. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)
23 pages, 968 KiB  
Review
Advanced Paternal Age in Focus: Unraveling Its Influence on Assisted Reproductive Technology Outcomes
by Aris Kaltsas, Athanasios Zikopoulos, Dionysios Vrachnis, Chara Skentou, Evangelos N. Symeonidis, Fotios Dimitriadis, Sofoklis Stavros, Michael Chrisofos, Nikolaos Sofikitis, Nikolaos Vrachnis and Athanasios Zachariou
J. Clin. Med. 2024, 13(10), 2731; https://doi.org/10.3390/jcm13102731 - 7 May 2024
Cited by 4 | Viewed by 2053
Abstract
As global demographics shift toward increasing paternal age, the realm of assisted reproductive technologies (ARTs), particularly in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), faces new challenges and opportunities. This study provides a comprehensive exploration of the implications of advanced paternal age [...] Read more.
As global demographics shift toward increasing paternal age, the realm of assisted reproductive technologies (ARTs), particularly in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), faces new challenges and opportunities. This study provides a comprehensive exploration of the implications of advanced paternal age on ART outcomes. Background research highlights the social, cultural, and economic factors driving men toward later fatherhood, with a focus on the impact of delayed paternity on reproductive outcomes. Methods involve a thorough review of existing literature, centering on changes in testicular function, semen quality, and genetic and epigenetic shifts associated with advancing age. Study results point to intricate associations between the father’s age and ART outcomes, with older age being linked to diminished semen quality, potential genetic risks, and varied impacts on embryo quality, implantation rates, and birth outcomes. The conclusions drawn from the current study suggest that while advanced paternal age presents certain risks and challenges, understanding and mitigating these through strategies such as sperm cryopreservation, lifestyle modifications, and preimplantation genetic testing can optimize ART outcomes. Future research directions are identified to further comprehend the epigenetic mechanisms and long-term effects of the older father on offspring health. This study underscores the need for a comprehensive approach in navigating the intricacies of delayed fatherhood within the context of ART, aiming for the best possible outcomes for couples and their children. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)
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22 pages, 321 KiB  
Review
Culture of Human Embryos at High and Low Oxygen Levels
by Ourania Konstantogianni, Theodoros Panou, Athanasios Zikopoulos, Charikleia Skentou, Sofoklis Stavros and Byron Asimakopoulos
J. Clin. Med. 2024, 13(8), 2222; https://doi.org/10.3390/jcm13082222 - 11 Apr 2024
Cited by 4 | Viewed by 1721
Abstract
One of the parameters potentially affecting the in vitro growth of preimplantation embryos is the oxygen concentration in the culture environment. An increased oxygen concentration causes the generation of ROS which in turn can cause damage to the cells and seriously disrupt the [...] Read more.
One of the parameters potentially affecting the in vitro growth of preimplantation embryos is the oxygen concentration in the culture environment. An increased oxygen concentration causes the generation of ROS which in turn can cause damage to the cells and seriously disrupt the embryonic development. Previous studies have assessed oxygen concentrations in the fallopian tubes of several mammals of between 5 and 8%, while the oxygen levels in the uterus were found to be even lower; similar measurements have been confirmed in humans. In addition, studies in mammalian embryos showed that low oxygen concentrations improve embryo development. Multiple studies on the effect of the oxygen concentration on human embryos have been conducted so far with diverse methodologies and objectives. Data from these have been included in three meta-analyses. All meta-analyses indicate the potential benefit in favor of a low oxygen concentration, though data are considered to be of a low methodological quality and further studies are considered necessary. However, based on the existing evidence, it is suggested that a low oxygen concentration should be adopted in the routine of the IVF laboratory, especially in the case of blastocyst culture. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)
20 pages, 1609 KiB  
Review
Evolution of Minimally Invasive and Non-Invasive Preimplantation Genetic Testing: An Overview
by Efthalia Moustakli, Athanasios Zikopoulos, Charikleia Skentou, Ioanna Bouba, Konstantinos Dafopoulos and Ioannis Georgiou
J. Clin. Med. 2024, 13(8), 2160; https://doi.org/10.3390/jcm13082160 - 9 Apr 2024
Cited by 1 | Viewed by 2303
Abstract
Preimplantation genetic testing (PGT) has become a common supplementary diagnοstic/testing tοol for in vitro fertilization (ΙVF) cycles due to a significant increase in cases of PGT fοr mοnogenic cοnditions (ΡGT-M) and de novο aneuplοidies (ΡGT-A) over the last ten years. This tendency is [...] Read more.
Preimplantation genetic testing (PGT) has become a common supplementary diagnοstic/testing tοol for in vitro fertilization (ΙVF) cycles due to a significant increase in cases of PGT fοr mοnogenic cοnditions (ΡGT-M) and de novο aneuplοidies (ΡGT-A) over the last ten years. This tendency is mostly attributable to the advancement and application of novel cytogenetic and molecular techniques in clinical practice that are capable of providing an efficient evaluation of the embryonic chromosomal complement and leading to better IVF/ICSI results. Although PGT is widely used, it requires invasive biopsy of the blastocyst, which may harm the embryo. Non-invasive approaches, like cell-free DNA (cfDNA) testing, have lower risks but have drawbacks in consistency and sensitivity. This review discusses new developments and opportunities in the field of preimplantation genetic testing, enhancing the overall effectiveness and accessibility of preimplantation testing in the framework of developments in genomic sequencing, bioinformatics, and the integration of artificial intelligence in the interpretation of genetic data. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)
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16 pages, 264 KiB  
Review
The Contribution of Proteomics in Understanding Endometrial Protein Expression in Women with Recurrent Implantation Failure
by Anastasios Potiris, Eleni Alyfanti, Eirini Drakaki, Despoina Mavrogianni, Theodoros Karampitsakos, Pavlos Machairoudias, Spyridon Topis, Athanasios Zikopoulos, Chara Skentou, Periklis Panagopoulos, Peter Drakakis and Sofoklis Stavros
J. Clin. Med. 2024, 13(7), 2145; https://doi.org/10.3390/jcm13072145 - 8 Apr 2024
Viewed by 1771
Abstract
Recurrent implantation failure (RIF) poses a significant challenge in assisted reproductive technology (ART) outcomes. The endometrium plays a crucial role in embryo implantation, and its protein expression profile is integral in determining receptivity. Proteomics has emerged as a valuable tool in unraveling the [...] Read more.
Recurrent implantation failure (RIF) poses a significant challenge in assisted reproductive technology (ART) outcomes. The endometrium plays a crucial role in embryo implantation, and its protein expression profile is integral in determining receptivity. Proteomics has emerged as a valuable tool in unraveling the molecular intricacies underlying endometrial receptivity and RIF. The aim of the present review is to analyze the contribution of proteomics to the understanding of endometrial protein expression in women with RIF, based on the results of significant proteomic studies. Medline/Pubmed databases were searched using keywords pertaining to proteomics combined with terms related to RIF. 15 studies were included in the present review. Several proteins have been found to exbibit differential expression in endometrial biopsies and fluid samples between fertile women and women with RIF during the receptive endometrial phase. The profile of endometrial proteins varied significantly among the studies. Nevertheless, similar changes in the expression levels of annexin-6, progesterone receptor, MMP-2, and MMP-9 in the endometrium of women with RIF, were found in more than one study indicating that certain proteins could potentially be effective biomarkers of endometrial receptivity. Proteomics contributes significantly to the understanding of protein expression in the endometrium of women with RIF and the analysis of proteins in endometrial fluid are promising for improving the clinical management of RIF. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)
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20 pages, 4116 KiB  
Review
Comprehensive Review of In Vitro Human Follicle Development for Fertility Restoration: Recent Achievements, Current Challenges, and Future Optimization Strategies
by Francisco Vitale and Marie-Madeleine Dolmans
J. Clin. Med. 2024, 13(6), 1791; https://doi.org/10.3390/jcm13061791 - 20 Mar 2024
Cited by 3 | Viewed by 1936
Abstract
Ovarian tissue cryopreservation (OTC) and subsequent transplantation (OTT) is a fertility preservation technique widely offered to prepubertal girls and young fertile women who need to undergo oncological treatment but are at a high risk of infertility. However, OTT is not considered safe in [...] Read more.
Ovarian tissue cryopreservation (OTC) and subsequent transplantation (OTT) is a fertility preservation technique widely offered to prepubertal girls and young fertile women who need to undergo oncological treatment but are at a high risk of infertility. However, OTT is not considered safe in patients with certain diseases like leukemia, Burkitt’s lymphoma, and ovarian cancer because of the associated risk of malignant cell reintroduction. In vitro follicle development has therefore emerged as a promising means of obtaining mature metaphase II (MII) oocytes from the primordial follicle (PMF) pool contained within cryopreserved ovarian tissue, without the need for transplantation. Despite its significant potential, this novel approach remains highly challenging, as it requires replication of the intricate process of intraovarian folliculogenesis. Recent advances in multi-step in vitro culture (IVC) systems, tailored to the specific needs of each follicle stage, have demonstrated the feasibility of generating mature oocytes (MII) from early-stage human follicles. While significant progress has been made, there is still room for improvement in terms of efficiency and productivity, and a long way to go before this IVC approach can be implemented in a clinical setting. This comprehensive review outlines the most significant improvements in recent years, current limitations, and future optimization strategies. Full article
(This article belongs to the Special Issue Challenges in Diagnosis and Treatment of Infertility)
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