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Advanced Techniques in Reproductive Medicine Research
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Advanced Techniques in Reproductive Medicine Research

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Biology".

Deadline for manuscript submissions: closed (31 July 2021) | Viewed by 24717

Special Issue Editors

Dr. Lorella Pascolo

E-Mail Website
Guest Editor
Institute for Maternal and Child Health, IRCCS Burlo Garofolo, 34137 Trieste, Italy
Interests: elementomics; iron-dyshomeostasis diseases; reproductive toxicology; endometriosis; nanomedicine; asbestos and nano-toxicology; membrane transport mechanisms; advanced imaging techniques
Special Issues, Collections and Topics in MDPI journals
Prof. Dr. Giuseppe Ricci

E-Mail Website
Guest Editor
1. Institute for Maternal and Child Health, IRCCS Burlo Garofolo, 34137 Trieste, Italy
2. Department of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, Italy
Interests: reproductive medicine; endometriosis; assisted reproduction techniques; reproductive biology; infertility diagnostics; fertility preservation; oncofertility
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Over the last 30 years, reproductive medicine has witnessed extraordinary scientific advances for infertility treatments, from in vitro fertilization (IVF) and successful embryo implantations to cryopreservation of gametes and embryos as well as ovary tissue cryopreservation for young cancer patients. Further advances are expected, since maintaining the reproductive health of individuals is nowadays of increased prominence, because many factors in industrialized countries seem to have resulted in an augmented incidence of infertility also among the young population. Meanwhile, research in reproductive medicine is becoming a multidisciplinary field, combining molecular biology techniques with new advanced approaches aimed at investigating the cause of reproductive failure and related diseases, or exploring the development of new diagnostics or therapeutics as well as exceptional challenges like the artificial ovary.

This issue invites research that aims at unravelling the molecular mechanisms underlying female and male human reproduction in addition to the use of unconventional and novel technical approaches that will help to preserve, promote, or restore healthy reproductive function.

We are most interested in technical methods dealing with X-ray techniques, vibrational spectroscopy, molecular imaging, omic technologies, AFM, and other advanced microscopies, as well as avantgarde therapeutic and diagnostic approaches like nanomedicine and nanotechnology, photobiomodulation therapy, microfluidics, and lab-on-chip.

Dr. Lorella Pascolo
Prof. Dr. Giuseppe Ricci
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • preservation of fertility
  • gamete quality
  • advanced techniques
  • ovary tissue
  • reproductive diseases
  • infertility
  • diagnosis and therapy

Related Special Issue

Published Papers (7 papers)

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Research

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29 pages, 8890 KiB  
Article
Nanosecond Pulsed Electric Field Only Transiently Affects the Cellular and Molecular Processes of Leydig Cells
by Wiktoria Kasprzycka, Alicja Trębińska-Stryjewska, Rafał Bogdan Lewandowski, Małgorzata Stępińska, Paulina Natalia Osuchowska, Monika Dobrzyńska, Yahia Achour, Łukasz Paweł Osuchowski, Jacek Starzyński, Zygmunt Mierczyk and Elżbieta Anna Trafny
Int. J. Mol. Sci. 2021, 22(20), 11236; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms222011236 - 18 Oct 2021
Cited by 4 | Viewed by 2007
Abstract
The purpose of this study was to verify whether the nanosecond pulsed electric field, not eliciting thermal effects, permanently changes the molecular processes and gene expression of Leydig TM3 cells. The cells were exposed to a moderate electric field (80 quasi-rectangular shape pulses, [...] Read more.
The purpose of this study was to verify whether the nanosecond pulsed electric field, not eliciting thermal effects, permanently changes the molecular processes and gene expression of Leydig TM3 cells. The cells were exposed to a moderate electric field (80 quasi-rectangular shape pulses, 60 ns pulse width, and an electric field of 14 kV/cm). The putative disturbances were recorded over 24 h. After exposure to the nanosecond pulsed electric field, a 19% increase in cell diameter, a loss of microvilli, and a 70% reduction in cell adhesion were observed. Some cells showed the nonapoptotic externalization of phosphatidylserine through the pores in the plasma membrane. The cell proportion in the subG1 phase increased by 8% at the expense of the S and G2/M phases, and the DNA was fragmented in a small proportion of the cells. The membrane mitochondrial potential and superoxide content decreased by 37% and 23%, respectively. Microarray’s transcriptome analysis demonstrated a negative transient effect on the expression of genes involved in oxidative phosphorylation, DNA repair, cell proliferation, and the overexpression of plasma membrane proteins. We conclude that nanosecond pulsed electric field affected the physiology and gene expression of TM3 cells transiently, with a noticeable heterogeneity of cellular responses. Full article
(This article belongs to the Special Issue Advanced Techniques in Reproductive Medicine Research)
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22 pages, 4597 KiB  
Article
Altered Follicular Fluid Metabolic Pattern Correlates with Female Infertility and Outcome Measures of In Vitro Fertilization
by Giacomo Lazzarino, Romina Pallisco, Gabriele Bilotta, Ilaria Listorti, Renata Mangione, Miriam Wissam Saab, Giuseppe Caruso, Angela Maria Amorini, Maria Violetta Brundo, Giuseppe Lazzarino, Barbara Tavazzi and Pasquale Bilotta
Int. J. Mol. Sci. 2021, 22(16), 8735; https://doi.org/10.3390/ijms22168735 - 14 Aug 2021
Cited by 15 | Viewed by 3120
Abstract
Nearly 40–50% of infertility problems are estimated to be of female origin. Previous studies dedicated to the analysis of metabolites in follicular fluid (FF) produced contrasting results, although some valuable indexes capable to discriminate control groups (CTRL) from infertile females (IF) and correlate [...] Read more.
Nearly 40–50% of infertility problems are estimated to be of female origin. Previous studies dedicated to the analysis of metabolites in follicular fluid (FF) produced contrasting results, although some valuable indexes capable to discriminate control groups (CTRL) from infertile females (IF) and correlate with outcome measures of assisted reproduction techniques were in some instances found. In this study, we analyzed in blind FF of 35 control subjects (CTRL = patients in which inability to obtain pregnancy was exclusively due to a male factor) and 145 IF (affected by: endometriosis, n = 19; polycystic ovary syndrome, n = 14; age-related reduced ovarian reserve, n = 58; reduced ovarian reserve, n = 29; unexplained infertility, n = 14; genetic infertility, n = 11) to determine concentrations of 55 water- and fat-soluble low molecular weight compounds (antioxidants, oxidative/nitrosative stress-related compounds, purines, pyrimidines, energy-related metabolites, and amino acids). Results evidenced that 27/55 of them had significantly different values in IF with respect to those measured in CTRL. The metabolic pattern of these potential biomarkers of infertility was cumulated (in both CTRL and IF) into a Biomarker Score index (incorporating the metabolic anomalies of FF), that fully discriminated CTRL (mean Biomarker Score value = 4.00 ± 2.30) from IF (mean Biomarker Score value = 14.88 ± 3.09, p < 0.001). The Biomarker Score values were significantly higher than those of CTRL in each of the six subgroups of IF. Posterior probability curves and ROC curve indicated that values of the Biomarker Score clustered CTRL and IF into two distinct groups, based on the individual FF metabolic profile. Furthermore, Biomarker Score values correlated with outcome measures of ovarian stimulation, in vitro fertilization, number and quality of blastocysts, clinical pregnancy, and healthy offspring. These results strongly suggest that the biochemical quality of FF deeply influences not only the effectiveness of IVF procedures but also the following embryonic development up to healthy newborns. The targeted metabolomic analysis of FF (using empowered Redox Energy Test) and the subsequent calculation of the Biomarker Score evidenced a set of 27 low molecular weight infertility biomarkers potentially useful in the laboratory managing of female infertility and to predict the success of assisted reproduction techniques. Full article
(This article belongs to the Special Issue Advanced Techniques in Reproductive Medicine Research)
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18 pages, 4242 KiB  
Article
Dysregulation of the Acrosome Formation Network by 8-oxoguanine (8-oxoG) in Infertile Sperm: A Case Report with Advanced Techniques
by Sung Woo Kim, Bongki Kim, Jongsoo Mok, Eun Seo Kim and Joonghoon Park
Int. J. Mol. Sci. 2021, 22(11), 5857; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22115857 - 30 May 2021
Cited by 2 | Viewed by 2482
Abstract
8-Hydroxyguanine (8-oxoG) is the most common oxidative DNA lesion and unrepaired 8-oxoG is associated with DNA fragmentation in sperm. However, the molecular effects of 8-oxoG on spermatogenesis are not entirely understood. Here, we identified one infertile bull (C14) due to asthenoteratozoospermia. We compared [...] Read more.
8-Hydroxyguanine (8-oxoG) is the most common oxidative DNA lesion and unrepaired 8-oxoG is associated with DNA fragmentation in sperm. However, the molecular effects of 8-oxoG on spermatogenesis are not entirely understood. Here, we identified one infertile bull (C14) due to asthenoteratozoospermia. We compared the global concentration of 8-oxoG by reverse-phase liquid chromatography/mass spectrometry (RP-LC/MS), the genomic distribution of 8-oxoG by next-generation sequencing (OG-seq), and the expression of sperm proteins by 2-dimensional polyacrylamide gel electrophoresis followed by peptide mass fingerprinting (2D-PAGE/PMF) in the sperm of C14 with those of a fertile bull (C13). We found that the average levels of 8-oxoG in C13 and C14 sperm were 0.027% and 0.044% of the total dG and it was significantly greater in infertile sperm DNA (p = 0.0028). Over 81% of the 8-oxoG loci were distributed around the transcription start site (TSS) and 165 genes harboring 8-oxoG were exclusive to infertile sperm. Functional enrichment and network analysis revealed that the Golgi apparatus was significantly enriched with the products from 8-oxoG genes of infertile sperm (q = 2.2 × 10−7). Proteomic analysis verified that acrosome-related proteins, including acrosin-binding protein (ACRBP), were downregulated in infertile sperm. These preliminary results suggest that 8-oxoG formation during spermatogenesis dysregulated the acrosome-related gene network, causing structural and functional defects of sperm and leading to infertility. Full article
(This article belongs to the Special Issue Advanced Techniques in Reproductive Medicine Research)
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16 pages, 2669 KiB  
Article
FTIR Spectroscopy to Reveal Lipid and Protein Changes Induced on Sperm by Capacitation: Bases for an Improvement of Sample Selection in ART
by Maria Pachetti, Luisa Zupin, Irene Venturin, Elisa Mitri, Rita Boscolo, Francesco D’Amico, Lisa Vaccari, Sergio Crovella, Giuseppe Ricci and Lorella Pascolo
Int. J. Mol. Sci. 2020, 21(22), 8659; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228659 - 17 Nov 2020
Cited by 11 | Viewed by 3422
Abstract
Although being a crucial step for Assisted Reproduction Technologies (ART) success, to date sperm selection is based only on morphology, motility and concentration characteristics. Considering the many possible alterations, there is a great need for analytical approaches allowing more effective sperm selections. The [...] Read more.
Although being a crucial step for Assisted Reproduction Technologies (ART) success, to date sperm selection is based only on morphology, motility and concentration characteristics. Considering the many possible alterations, there is a great need for analytical approaches allowing more effective sperm selections. The use of Fourier Transform Infrared (FTIR) may represent an interesting possibility, being able to reveal many macromolecular changes in a single measurement in a nondestructive way. As a proof of concept, in this observational study, we used a FTIR approach to reveal features related to sperm quality and chemical changes promoted by in vitro capacitation. We found indication that α-helix content is increased in capacitated sperm, while high percentages of the β-structures seem to correlate to poor-quality spermatozoa. The most interesting observation was related to the lipid composition, when measured as CH2/CH3 vibrations (ratio 2853/2870), which resulted in being strongly influenced by capacitation and well correlated with sperm motility. Interestingly, this ratio is higher than 1 in infertile samples, suggesting that motility is related to sperm membranes stiffness and lipid composition. Although further analyses are requested, our results support the concept that FTIR can be proposed as a new smart diagnostic tool for semen quality assessment in ART. Full article
(This article belongs to the Special Issue Advanced Techniques in Reproductive Medicine Research)
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Review

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17 pages, 2110 KiB  
Review
Scanning Probe Microscopies: Imaging and Biomechanics in Reproductive Medicine Research
by Laura Andolfi, Alice Battistella, Michele Zanetti, Marco Lazzarino, Lorella Pascolo, Federico Romano and Giuseppe Ricci
Int. J. Mol. Sci. 2021, 22(8), 3823; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22083823 - 07 Apr 2021
Cited by 3 | Viewed by 2466
Abstract
Basic and translational research in reproductive medicine can provide new insights with the application of scanning probe microscopies, such as atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM). These microscopies, which provide images with spatial resolution well beyond the optical resolution [...] Read more.
Basic and translational research in reproductive medicine can provide new insights with the application of scanning probe microscopies, such as atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM). These microscopies, which provide images with spatial resolution well beyond the optical resolution limit, enable users to achieve detailed descriptions of cell topography, inner cellular structure organization, and arrangements of single or cluster membrane proteins. A peculiar characteristic of AFM operating in force spectroscopy mode is its inherent ability to measure the interaction forces between single proteins or cells, and to quantify the mechanical properties (i.e., elasticity, viscoelasticity, and viscosity) of cells and tissues. The knowledge of the cell ultrastructure, the macromolecule organization, the protein dynamics, the investigation of biological interaction forces, and the quantification of biomechanical features can be essential clues for identifying the molecular mechanisms that govern responses in living cells. This review highlights the main findings achieved by the use of AFM and SNOM in assisted reproductive research, such as the description of gamete morphology; the quantification of mechanical properties of gametes; the role of forces in embryo development; the significance of investigating single-molecule interaction forces; the characterization of disorders of the reproductive system; and the visualization of molecular organization. New perspectives of analysis opened up by applying these techniques and the translational impacts on reproductive medicine are discussed. Full article
(This article belongs to the Special Issue Advanced Techniques in Reproductive Medicine Research)
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13 pages, 502 KiB  
Review
The Amniotic Fluid Cell-Free Transcriptome Provides Novel Information about Fetal Development and Placental Cellular Dynamics
by Hee Jin Park, Hee Young Cho and Dong Hyun Cha
Int. J. Mol. Sci. 2021, 22(5), 2612; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22052612 - 05 Mar 2021
Cited by 9 | Viewed by 3888
Abstract
The amniotic fluid (AF) is a complex biofluid that reflects fetal well-being during development. AF con be divided into two fractions, the supernatant and amniocytes. The supernatant contains cell-free components, including placenta-derived microparticles, protein, cell-free fetal DNA, and cell-free fetal RNA from the [...] Read more.
The amniotic fluid (AF) is a complex biofluid that reflects fetal well-being during development. AF con be divided into two fractions, the supernatant and amniocytes. The supernatant contains cell-free components, including placenta-derived microparticles, protein, cell-free fetal DNA, and cell-free fetal RNA from the fetus. Cell-free mRNA (cfRNA) analysis holds a special position among high-throughput analyses, such as transcriptomics, proteomics, and metabolomics, owing to its ease of profiling. The AF cell-free transcriptome differs from the amniocyte transcriptome and alters with the progression of pregnancy and is often associated with the development of various organ systems including the fetal lung, skin, brain, pancreas, adrenal gland, gastrointestinal system, etc. The AF cell-free transcriptome is affected not only by normal physiologies, such as fetal sex, gestational age, and fetal maturity, but also by pathologic mechanisms such as maternal obesity, and genetic syndromes (Down, Edward, Turner, etc.), as well as pregnancy complications (preeclampsia, intrauterine growth restriction, preterm birth, etc.). cfRNA in the amniotic fluid originates from the placenta and fetal organs directly contacting the amniotic fluid as well as from the fetal plasma across the placenta. The AF transcriptome may reflect the fetal and placental development and therefore aid in the monitoring of normal and abnormal development. Full article
(This article belongs to the Special Issue Advanced Techniques in Reproductive Medicine Research)
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9 pages, 266 KiB  
Review
Ovarian Aging: Molecular Mechanisms and Medical Management
by Jan Tesarik, Maribel Galán-Lázaro and Raquel Mendoza-Tesarik
Int. J. Mol. Sci. 2021, 22(3), 1371; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22031371 - 29 Jan 2021
Cited by 41 | Viewed by 6130
Abstract
This is a short review of the basic molecular mechanisms of ovarian aging, written with a particular focus on the use of this data to improve the diagnostic and therapeutic protocols both for women affected by physiological (age-related) ovarian decay and for those [...] Read more.
This is a short review of the basic molecular mechanisms of ovarian aging, written with a particular focus on the use of this data to improve the diagnostic and therapeutic protocols both for women affected by physiological (age-related) ovarian decay and for those suffering premature ovarian insufficiency. Ovarian aging has a genetic basis that conditions the ovarian activity via a plethora of cell-signaling pathways that control the functions of different types of cells in the ovary. There are various factors that can influence these pathways so as to reduce their efficiency. Oxidative stress, often related to mitochondrial dysfunction, leading to the apoptosis of ovarian cells, can be at the origin of vicious circles in which the primary cause feeds back other abnormalities, resulting in an overall decline in the ovarian activity and in the quantity and quality of oocytes. The correct diagnosis of the molecular mechanisms involved in ovarian aging can serve to design treatment strategies that can slow down ovarian decay and increase the quantity and quality of oocytes that can be obtained for an in vitro fertilization attempt. The available treatment options include the use of antioxidants, melatonin, growth hormones, and mitochondrial therapies. All of these treatments have to be considered in the context of each couple’s history and current clinical condition, and a customized (patient-tailored) treatment protocol is to be elaborated. Full article
(This article belongs to the Special Issue Advanced Techniques in Reproductive Medicine Research)
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