Functional Ovarian Tissue Research Articles

7709 Ovotesticular Difference of Sex Development: Prognostic Evaluation, Sex Assignment, and Ethics

Abstract Disclosure: X. Xu: None. Y. Lin: None. B. Shivanna: None. A. Lee: None. J.C. Hakim: None. D.G. Mann: None. L.M. Noll: None. P. Georgiadis: None. S.K. Gunn: None. L.P. Karaviti: None. Background: Our case provides the identification and management of a newborn baby with ovotesticular syndrome, which was formerly known as true hermaphroditism. The term was classically derived from Greek mythology in which the deity 'Hermaphroditus', merging with the nymph Salamis, embodied both male and female attributes. The term 'true hermaphroditism' is now avoided due to its stigmatizing connotations. Ovotesticular difference of sex development (DSD) is an exceedingly rare condition, occurring in fewer than 1 in 20,000 births. This condition involves the coexistence of functional ovarian and testicular tissue within one individual, presenting a challenge in sex assignment due to unpredictable gender development. Clinical Case: This is a single case of the presence of ovarian and testicular differentiation on one side and a transitional zone of mixed pattern. Prenatal genetic testing consistent 46,XX karyotype. Initial ultrasound at the outside hospital revealed a uterus and right testicle. The baby exhibited Prader stage 4 genitalia, with a palpable gonad in the right labioscrotal area and the left gonad not palpable. Prophylactic hydrocortisone was started due to concerns regarding NR5A1/SF1 mutation. Further evaluations, including genetic tests, hormonal assessments, urinalysis and kidney ultrasound for the WT1 mutation, and scrotal and pelvic ultrasounds, were conducted. The high-dose ACTH stimulation test ruled out adrenal insufficiency. The testosterone level was in the male range, and the HCG stimulation test further confirmed functioning Leydig cells. The postnatal karyotype was 46,XX, and chromosome microarray analysis showed no abnormalities. Our approach to this case was multidisciplinary, including our endocrine experts, urologist, gynecologist, ethicist, and psychologist. We discussed the fluidity of sex assignment and addressed the degree of virilization, nature of the gonadal tissue, and emphasized the necessity of regular clinical follow-up. Our main concern was to educate the parents and protect the choices of the child. In this case, the child assumed a male sex assignment, as a result of the decision-making process between our team and the parents. In this context we did emphasize the fluidity of sex assignment. Clinical Lesson: Long-term outcomes for ovotesticular DSD remain understudied. Existing reports highlight varying rates of gender dysphoria, sexual function, and psychological well-being. Our approach to managing ovotesticular DSD involved a framework emphasizing ethical considerations and parental involvement. We advocated for and practiced a shared decision-making model involving healthcare providers, parents, and ultimately the patient. Presentation: 6/3/2024

Repair effect of human umbilical cord mesenchymal stem cell-derived small extracellular vesicles on ovarian injury induced by cisplatin.

Small extracellular vesicles (sEVs) secreted by human umbilical cord have therapeutic effects on various degenerative diseases. However, the characteristics and potential functions of human umbilical cord mesenchymal stem cells (huMSCs)-derived sEVs, especially the role of premature ovarian failure (POF), are poorly understood. Here, we isolated and characterized huMSCs and their sEVs. huMSCs highly expressed CD73, CD90, and CD105. huMSC-sEVs showed typical exosomal features, highly expressing CD9, TSG101, and CD63. It was shown that huMSC-sEVs could be taken up by granulosa cells (GCs) and damaged ovarian tissue, which increased the levels of hormone secretion and reduced GCs apoptosis. We further confirmed that the levels of follicle-stimulating hormone in rat serum decreased dramatically, while the levels of estrogen (E2）and anti-mullerian hormone (AMH) increased significantly with the treatment of huMSC-sEVs. Meanwhile, huMSC-sEVs treatment greatly reduced cell apoptosis and autophagy, while increased the phosphorylation levels of p-PI3K and p-Akt. Therefore, treatment with huMSC-sEVs significantly inhibited GCs apoptosis, improved ovarian morphology, promoted follicular development, inhibited follicular over-atresia, and improved ovarian reserve capacity in POF rats. Our study verified that activation of PI3K/Akt signaling pathway and regulation of cellular autophagy, thus reducing GCs death, are the mechanisms by which huMSC-sEVs restore ovarian tissue function.

Engineering of ovarian tissue for ovarian dysfunctions: A review

This review explores tissue engineering as a potential solution for reproductive health issues in women caused by genetic or acquired diseases, such as premature ovarian failure or oophorectomy. The loss of ovarian function can lead to infertility, osteoporosis, and cardiovascular disease. Hormone replacement therapy is a common treatment, but it has limitations and risks. The review focuses on two main approaches in tissue engineering: scaffold-based (3D printing, electrospinning, decellularization) and scaffold-free (stem cell transplantation, organoid cultivation). Both approaches show promise in preclinical studies for creating functional ovarian tissue. Challenges include vascularization, innervation, long-term function, and safety. Despite these challenges, tissue engineering offers a potential avenue for restoring fertility and hormone balance in women with ovarian dysfunction.

OR19-06 State-of-the-art MALDI Mass Spectrometry Imaging Reveals Unique Spatial Lipid Signatures in Human and Mouse Ovarian Compartments

Abstract Disclosure: N. Pascuali: None. F. Tobias: None. K. Valyi-Nagy: None. S. Salih: None. A. Veiga-Lopez: None. The incidence of infertility has been consistently increasing over the past few decades, with a high proportion of these cases caused by ovarian-related dysfunction. Metabolic disorders such as hyper- or dyslipidemia have been associated with subfertility (e.g., follicular fluid dyslipidemia is associated with lower IVF success). Lipids play a critical role in the structure and function of ovarian tissue. However, while several studies have investigated transcript- and protein-level alterations in ovarian pathologies, less is known about alterations in the lipid landscape. The first step in determining if ovarian disorders are associated with ovarian dyslipidemia - and thus use lipids as potential biomarkers of ovarian pathology - is to understand the physiological distribution of ovarian lipids. Matrix-Assisted Laser Desorption Ionization Time-Of-Flight (MALDI-TOF) mass spectrometry imaging (MSI) enables label-free in situ analysis of molecules such as lipids and metabolites directly within tissue sections. We aimed to begin mapping the distribution of lipids in human and mouse ovaries using high spatial resolution MALDI MSI. To this end, and after informed consent, two fresh ovarian biopsies from a 26-year-old patient were frozen, cryosectioned into 10-μm sections, and mounted onto ITO-coated slides. A set of 21-day-old mouse ovaries (n=10) were gelatin-embedded and cryosectioned. Slides were sprayed with 1,5-diaminonaphthalene as the matrix. MALDI MSI was performed using a Bruker RapifleX MALDI Tissuetyper (20-μm resolution) in the negative ion mode. Data were processed using the SCiLS Lab software. We identified for the first time in human ovaries over 100 lipid species, with most of them displaying distinct spatial distributions. Specifically, we detected 3 lipids specific to theca cells (m/z: 701.6; 1050.8; 1263.9), 6 lipids specific to granulosa cells (m/z: 279.3; 697.5; 861.7; 883.7; 911.7; 1448.0), 3 lipids specific to the stromal region (m/z 392.1; 753.5; 835.7), and one specific to luteal cells (m/z: 615.3). As for mouse ovaries, we detected 5 lipids specific to follicular cells (theca or granulosa) (m/z: 327.1: 343.2; 552.6; 632.6; 703.5), 5 lipids specific to stromal cells (m/z: 715.6; 729.6; 743.6; 771.7; 799.6), 5 lipids specific to oocytes (m/z: 778.5; 863.7; 1063.8; 1225.7; 1336.0) and 7 lipids specific to the ovarian cortex (m/z: 721.5; 748.5; 776.6; 792.6; 810.6; 869.6; 885.7). To our knowledge, this is the first evaluation of the human ovarian lipidome and characterization of region- or cell-layer-specific lipids. This approach has allowed us to demonstrate that compartments as small as oocytes (∼70 μm in diameter) have a unique lipid signature. Harnessing the unique specificity of the lipid signature per ovarian region and/or cell layer in this study could help pave the way to develop novel biomarkers of ovarian dysfunction. Presentation Date: Saturday, June 17, 2023

Comparison the therapeutic effects of bone marrow CD144+ endothelial cells and CD146+ mesenchymal stem cells in POF rats.

Premature ovarian insufficiency (POI) is a challenging issue in terms of reproduction biology. In this study, therapeutic properties of bone marrow CD146+ mesenchymal stem cells (MSCs) and CD144+ endothelial cells (ECs) were separately investigated in rats with POI. POI rats were classified into control POI, POI + CD146+ MSCs, and POI + CD144+ ECs groups. Enriched CD146+ MSCs and CD144+ ECs were directly injected into ovarian tissue (15 × 104 cells/10 μL) in relevant groups. After 4 weeks, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) levels were measured in blood samples. Ovarian tissues were collected and subjected to Hematoxylin-Eosin and Masson's trichrome staining. The expression of angp-2, vegfr-2, smad-2, -4, -6, and tgf-β1 was studied using qRT-PCR analysis. Histopathological examination indicated an increased pattern of atretic follicles in the POI group related to the control rats (P<0.0001). Data indicated that injection of POI + CD146+ MSCs and CD144+ ECs in POI rats reduced atretic follicles and increased the number of normal follicles (P<0.01). Along with these changes, the content of blue-colored collagen fibers was diminished after cell transplantation. Besides, cell transplantation in POI rats had the potential to reduce increased FSH, and LH levels (P<0.05). In contrast, E2 content was increased in POI + CD146+ MSCs and POI + CD144+ ECs groups compared to control POI rats, indicating restoration of follicular function. CD144+ (smad-2, and -4) and CD146+ (smad-6) cells altered the activity of genes belonging TGF-β signaling pathway. Unlike POI + CD146+ MSCs, aberrant angiogenesis properties were significantly down-regulated in POI + CD144+ ECs related to the control POI group (P<0.05). The transplantation of bone marrow CD146+ and CD144+ cells can lead to the restoration of ovarian tissue function in POI rats via modulating different mechanisms associated with angiogenesis and fibrosis.

Ovarian remnant syndrome in small animals: case series

Our objective was to report the utility of various diagnostic tests in identifying ovarian remnant syndrome (ORS) in dogs and cats. Medical records from 2 referral teaching hospitals (Health Sciences Centre at the Ontario Veterinary College and Cornell University Hospital for Animals) were examined and 48 animals (31 dogs and 17 cats) were chosen. Data included were based on sufficient clinical or diagnostic evidence of ORS. Histopathology was used as the confirmatory test for ORS. There was no difference between the proportions of dogs versus cats diagnosed with ORS. Similarly, there was no difference in the proportions of ovarian remnants (OR) between large, medium, or small dogs, or the side (right, left, or bilateral) in which OR was diagnosed. Vaginal cytology and transabdominal ultrasonography findings, and serum progesterone concentrations had the highest chance of correctly identifying an OR prior to exploratory surgery. Transabdominal ultrasonography had strong agreement with OR location identified at surgery. Presumptive intraoperative diagnosis of OR was possible in 39/41 cases (95.1%). Auxiliary diagnostic testing should be recommended to confirm functional ovarian tissue before surgery to reduce unnecessary surgery. Additionally, transabdominal ultrasonography examination may reduce surgical time since ovarian remnant location has strong agreement with surgical findings.

Development of the human ovary: Fetal through pubertal ovarian morphology, folliculogenesis and expression of cellular differentiation markers

A definition of normal human fetal and early postnatal ovarian development is critical to the ability to accurately diagnose the presence or absence of functional ovarian tissue in clinical specimens. Through assembling an extensive histologic and immunohistochemical developmental ontogeny of human ovarian specimens from 8 weeks of gestation through 16 years of postnatal, we present a comprehensive immunohistochemical mapping of normal protein expression patterns in the early fetal through post-pubertal human ovary and detail a specific expression-based definition of the early stages of follicular development. Normal fetal and postnatal ovarian tissue is defined by the presence of follicular structures and characteristic immunohistochemical staining patterns, including granulosa cells expressing Forkhead Box Protein L2 (FOXL2). However, the current standard array of immunohistochemical markers poorly defines ovarian stromal tissue, and additional work is needed to identify new markers to advance our ability to accurately identify ovarian stromal components in gonadal specimens from patients with disorders of sexual differentiation.

Ovarian remnant syndrome in feline: Case report

Ovarian remnant syndrome is characterized by the presence of functional ovarian tissue after ovariohysterectomy (OH). In this report, the case of a 2-year-old female cat who started to show estrus behaviors 10 months after OH, correlated with the absence of the tutor. The exams of choice to reach a first diagnosis were vaginal cytology and abdominal ultrasound. The designated treatment, was excision of the ovarian tissue fragment thought exploratory laparotomy. Histopathological examination confirmed the diagnosis.

Influence of hormonal stimulation and intermittent cold exposure on the mechanisms of formation of polycystic ovarian syndrome

Introduction. Polycystic ovary syndrome (PCOS) is one of the most pressing problems in gynecological endocrinology. The aim of the study was to elucidate the morphological changes in the ovaries of rats under various types of hormonal exposure in experimental modeling of PCOS. Materials and methods. To study three working experimental models were developed, associated with the introduction of various hormonal drugs into the body of an animal. Model I-st consisted of subcutaneous administration of an estrogen-containing drug at a dose of 3 mg for 25 days. Model II-d involved the development of PCOS by daily subcutaneous administration of 3 mg of dehydroandrostenediol sulfate. The III model - the formation of PCOS in 5 immature rats using dopamine phytoanalogues at a dose of 3 mg subcutaneously for 25 days. The animals were divided into 3 groups: group 1 - rats that were induced with experimental PCOS and kept in a vivarium at a temperature of +23°C (n=8), group 2 - animals that, against the background of experimental PCOS, were exposed to cold exposure (CE) for 4 hours in a chamber with a constant light regime and a temperature of + 4 °C for 25 days (n = 8); group 3 - intact control (n = 8) - healthy rats without PCOS, which were kept in a vivarium at a temperature of + 23°C. Results. Histological examination of the ovaries of group 1 rats revealed morphological changes in the ovaries characteristic of PCOS. The morphological structure of the ovaries corresponded to type II PCOS. In rats of group 3, macroscopically, the ovaries were slightly reduced in size compared to the control group. The levels of adiponectin and leptin as markers of the functional ovarian tissue of PCOS model experiments.When studying the effect of CE on the body of experimental animals, their effect on the formation of tissue cell hyperplasia as a characteristic sign of PCOS was determined. Conclusions. Studies have shown that the formation of PCOS, as well as structural changes in the ovaries, significantly depend on hormonal regulation disorders.

O-276 Reproductive outcomes and fertility-sparing in women with Borderline ovarian tumors

Abstract The standard treatment of BOT is defined as a total abdominal hysterectomy and bilateral salpingo-oophorectomy, peritoneal cytology, omentectomy and multiple peritoneal biopsies. These procedures allowed to perform an adequate staging, most of the time no adjuvant treatment is necessary except in case of associated invasive peritoneal implants. The prognosis of BOT is excellent. However, late recurrences (after 5 or 10 years) can be observed. Conservative surgery is defined as preservation of the uterus and at least part of one ovary, in order to preserve fertility. BOT arise in a young population, for whom fertility is a major issue. The global recurrence risk is estimated at 13%. The rate of recurrence is correlated with the type of conservative treatment used (salpingo-oophorectomy or cystectomy) with a higher rate of between 10 and 42% in patients undergoing cystectomy. In the case of bilateral serous BOT, if technically feasible, bilateral cystectomy should be performed to improve pregnancy rates. For serous BOT e cystectomy can be performed (tumour can be bilateral for 20% of these patients) with a close follow up by ultrasound. For mucinous BOT, salpingo-oophorectomy is recommended to avoid rare but at risk invasive recurrence, (these lesions are usually unilateral). A complete evaluation must be performed preoperatively including MRI with evaluation of possible safe functional ovarian tissue and oncofertility consultation to discuss if fertility preservation technique can be proposed before surgery. The observed pregnancy rates observed after conservative surgery are between 32 and 88%. Different factors can influence fertility rates: type of conservative treatment, age of the patient (with almost a quarter of the live birth occurring after recurrence with no more further event to date, a second fertility-sparing surgery after local borderline recurrence can be proposed in the case of pregnancy project), histologic subtype of the tumour (better with mucinous versus serous), the use of a laparoscopic approach and the use of a 2- or 3-step surgery (initial, restaging, second look). In spite of conservative management in BOT, some patients will experience infertility. In vitro fertilization (IVF) procedures have not been significantly associated with a specific increase in BOT or ovarian cancer rates. In vitro data suggest that gonadotropins and/or high dose of estrogens don’t induce a borderline cell cultures proliferation. Analyzing all series reporting IVF and BOT, the pooled estimate for pregnancy was 80% (95% CI: 68–92%). The pooled estimate for recurrence was 23% (95% CI: 6–39%). The rate of recurrence in these women is ‘low’ though this is probably due to the fact that women who are selected as eligible for ART have a better prognosis and more often early-stage BOT. However, there is a real need for fertility preservation expert centers associating oncologists and fertility experts who can evaluate conservative management of BOT along with alternative therapeutic options to preserve fertility as well as ART.

Endocrine Function of Ovarian Tissue Grafts After Normo- and Hypothermic Storage

Morphological characteristics and function of ovarian tissue after 2–120 hrs of storage at 4 and 37°C in incubation media with different composition (mannitol-containing solution (MСS) and phosphate buffered saline (PBS)) were investigated. The function of ovarian tissue after normo- and hypothermic storage was studied by heterotopic transplantation to recipient animals simultaneously with bilateral ovariectomy. After 2-hour storage at 37ºC and 24-hour one at 4ºC in all the studied media, the estradiol and progesterone levels in plasma of recipient animals were experimentally proved to be 18.2–27.5 and 13.5–18.5 ng/ml, respectively, (after fresh tissue transplantation these values for estradiol and progesterone made (28.1 ± 3.6) pg/ml and (19.1 ± 2.8) ng/ml, respectively). Prolongation of tissue storage time at 37ºC increased the number of degenerative follicles, augmented TBARS concentration and reduced sex hormone levels after transplantation using both MCS and PBS. A protective effect of MCS only, manifested in endocrine function restoration in recipient animals after transplantation ((13.5 ± 2.5) pg/ml of estradiol) was found after 48 hrs of ovarian tissue hypothermic storage. If PBS was used, the estradiol level was within the indices for ovariectomized animals ((7.1 ± 1.5 (pg/ml)).

Effects of AavLEA1 Protein on Mouse Ovarian Tissue Cryopreservation by Vitrification.

Conventional ovarian tissue cryopreservation often destroys the structural, functional, and DNA integrity of the ovarian tissue. How to effectively retain the ultrastructure and subsequent function of ovarian tissue during cryopreservation has long been an issue of concern. Late embryogenesis abundant (LEA) proteins are a class of highly hydrophilic proteins and have been reported to protect various cells from water stress. However, whether LEA proteins exert protective effects on ovarian tissue cryopreservation remains unknown. To investigate the benefit of LEA proteins in ovarian tissue cryopreservation, we purified the recombinant AavLEA1 protein, a member of Group 3 LEA proteins, then cryopreserved the mouse ovaries with this protein by vitrification, and obtained the ovarian follicle structure, cellular proliferation, apoptosis, and GAPDH gene expression of postcryopreservation ovaries. We found that recombinant AavLEA1 protein protected the ovarian follicles from cryoinjury, improved the proliferative ability of follicles, decreased the apoptosis, and promoted the GAPDH gene expression. These results indicated that the LEA protein enhanced the antiapoptosis ability of ovarian cells and retained DNA/RNA integrity against cryoinjury during ovarian tissue vitrification. LEA proteins exert beneficial effects on ovarian tissue cryopreservation, and maybe provide a novel cryoprotective agent for ovarian tissue cryopreservation.

A clinicopathological study of ovarian endometriotic cysts

Endometriosis is an important gynecologic disorder with multifactorial causes, primarily affecting women during their reproductive years. Pathologically, it is the result of functional endometrium located outside the uterus which may vary from microscopic endometriotic implants to large cysts. Endometriotic cysts and infertility is a well-known association. Some patients are asymptomatic while others present with disabling pelvic pain, infertility, or adnexal masses. Cyst aspiration, fenestration and ablation of cyst wall are commonly performed surgical procedures. Excision of the cyst wall is an accepted surgical treatment owing to the low recurrence rates. A total of 35 patients who underwent ovarian cystectomy for endometriotic cysts between January 2019 and December 2020 were retrospectively identified. The clinical findings, gross and histopathological features were noted in each case. Microscopically, the presence or absence of ovarian tissue adjacent to the cyst wall was evaluated. If ovarian tissue was present, the morphologic characteristics were graded on a semi-quantitative scale of 0-4 as described by Muzii et al. The age group of patients ranged between 22-28yrs. Right side cysts accounted for the majority, however 6 cases had bilateral endometriotic cysts. Majority of patients presented with primary infertility (46.2%). The maximum weight recorded for these cysts was 35gm, size ranging between 4.5 to 18cm and median thickness of the cyst wall being 0.7cm. 68% of the cysts showed a lining epithelium, few showing atypia and oncocytic change. Fibrosis and hemosiderin laden macrophages were present in more than 70% of cases and endometrial glands and stroma in more than 50%. Inflammation when present was predominantly lymphocytic. On evaluation of the ovarian tissue, 42.8% of cases showed no follicles and the rest showing grades ranging from 1 to 4, with grade 1 accounting for majority. The present study further emphasizes endometriosis to be an important cause of primary infertility which needs to be recognized and treated appropriately. Recognition of these cysts on histopathological examination can be challenging at times when endometrial stroma is scant and in cases of tubo-ovarian masses where these lesions could mimic malignancy. The excision of endometriotic cyst wall may cause loss of functional ovarian tissue in patients with primary infertility and thus could effect the response to ovarian stimulation, ocyte recovery, implantation and fertilization rates in these patients.

P–436 Identification of ovarian cell subpopulations by multicolor flow cytometry and its potential impact on ovarian reconstruction programs

Abstract Study question How could multicolor flow cytometry (MFC) help to identify ovarian subpopulations that could be used for ovarian reconstruction with isolated follicles? Summary answer MFC is useful to identify ovarian cell subpopulations in the ovarian cortex. What is known already Ovarian tissue cryopreservation is a fertility preservation option for women before gonadotoxic chemo- and/or radiotherapy. However, graft of cryopreserved ovarian tissue must be performed with caution in women suffering from malignancies that may metastasize to the ovaries. For this purpose, functional ovarian tissue qualification is essential to identify ovarian cell subpopulations that could be used for ovary reconstruction in combination with isolated follicles. Furthermore, ischemic tissue damage occurring after the graft is currently another important issue to be resolved for successful ovarian reuse. Study design, size, duration We developed an automated ovarian tissue dissociation method to obtain ovarian cell suspensions. Then, we used MFC for the identification of different cell subpopulations in the cell suspension thus obtained. Participants/materials, setting, methods Human ovarian tissues from patients undergoing surgery for polycystic ovary syndrome were used in this study. Biopsies of ovarian cortex (fresh or frozen-thawed) were dissociated using an automated dissociation method. We used FVS780 and SYTO13 markers to gate viable ovarian cells by MFC. Variable markers were chosen to differentiate and identify cell subpopulations among the viable ovarian cells. Main results and the role of chance The dissociation yield was on average 1.59 ± 1.58 x 106 and 0.78 ± 0.72 x 106 viable ovarian cells per 100 mg of fresh (n = 17) and frozen-thawed (n = 43) ovarian cortical tissue, respectively. On average, 35.4 ± 13.1% of viable ovarian cells were CD34 + (n = 61, stromal phenotype). Concerning endothelial phenotype, 7.8 ± 5.5% of CD31+ cells (n = 51) and 5.3 ± 3.6% of CD144+ cells (n = 29) were identified among viable ovarian cells. Vimentin marker is found in 25.6 ± 10.8% of viable ovarian cells (n = 23) and CD326 (EpCAM expression) in 0.6 ± 0.8% (n = 16). Finally, pericyte phenotype (CD34-/Vimentin-/CD31-/CD146+/ CD140b+) was identified in 4.6 ± 4.3% of viable ovarian cells (n = 7). Limitations, reasons for caution We do not know how these ovarian cell subpopulations could be a factor associated or not with time for ovarian function recovery in vivo after ovarian tissue graft and the impact of these ovarian cells on the ovarian microenvironment of an artificial ovary. Wider implications of the findings: Functional qualification of ovarian tissue can be performed by MFC. MFC is a promising tool for ovarian cortex qualification before reuse of cryopreserved ovarian tissue. Cell sorting could be used to separate and isolate cell subpopulations and add these cells with isolated follicles in an ovarian reconstruction program. Trial registration number Not applicable

Intra-Ovarian Platelet-Rich Plasma Injections: Single-Centre Experience.

Objective: To describe the effects of intra-ovarian platelet-rich plasma injection on the ovarian stimulation outcomes in women referring to an in vitro fertilization center. Method: We conducted a single-center retrospective study on 179 women that underwent intra-ovarian platelet-rich plasma injection over the last three years. Inclusion criteria included women over age 35 with at least one ovary with a history of infertility, hormonal abnormalities, absence of menstrual cycle and premature ovarian failure. Results: Mean (±SD) patient age was 43 ± 4 years. Both serum FSH levels and serum E2 significantly reduced after treatment from 29.0 pg/ml to 18.0 pg/ml; p&lt;0.0001 and from 65.6 pg/ml to 47.2 pg/ml; p= 0.034 respectively. None of the 179 women reported any complications post operatively. After PRP, 17/179 (9.49%) women became pregnant. Conclusion: The results of our observational study revealed that PRP intra-οvarian injection is associated with improved function of ovarian tissue. Future further randomized clinical trials in this field are needed to shed light in the use of PRP in ovarian rejuvenation.

Spatiotemporal changes in mechanical matrisome components of the human ovary from prepuberty to menopause.

Spatiotemporal changes in mechanical matrisome components of the human ovary from prepuberty to menopause.

Uterine stump adenocarcinoma in a bitch with an ovarian remnant: A case report

A 3-year-old female spayed dog was presented with a history of sanguineous vaginal discharge of 2 month duration. The dog was spayed 1.5 years before presentation. Clinical examination revealed a large, solid, ovoid mass in the caudal abdomen, recognized by diagnostic imaging as an enlargement at the top of the uterine stump. Additionally, high serum progesterone concentration was measured, confirming the presence of functional ovarian tissue in dioestrus. With a suspicion for a related uterine stump pyometra or less likely, for other enlargements, a cοeliotomy was performed. A mass at the apex of the uterine body and a right side ovarian remnant were found. Both structures and the remaining uterine stump were excised. The uterine remnant mass was histologically diagnosed as uterine adenocarcinoma, herein detected for the first time at the uterine stump in the bitch. Unsuccessful, incomplete ovariohysterectomy may permit even neoplastic transformation of uterine stump tissues.

INDICATORS OF OVARIAL RESERVE, PRO- AND ANTIOXIDANT SYSTEM, CYTOKINE STATUS OF PERIPHERAL BLOOD AND PERITONEAL LIQUID IN PATIENTS WITH ENDOMETRIOID OVARIAN CYSTS

The aimof this work was to identify the characteristics of the cytokine status and state of the pro- and antioxidant systems in patients of late reproductive age with endometrioid cysts and determine their role in changing the ovarian reserve.Materials and methods.A total of 89 patients with endometrioid ovarian cysts (EOC) were examined. The control group consisted of 35 healthy patients admitted for surgical sterilisation. Peripheral blood was collected before surgery, while the peritoneal fluid collection was performed intraoperatively. In peripheral blood and peritoneal fluid, the level of cytokines was determined. The calculation of the anti-inflammatory index was performed, along with the evaluation of the content of lipid peroxidation products and the antioxidant activity. The analysis of the ovarian reserve parameters was conducted.Results.In patients of late reproductive age suffering from EOC, a decrease in the ovarian reserve is observed under the preserved hormonal background. The presence of EOC is accompanied by the activation of lipid peroxidation, both at the systemic and local levels, under a pronounced inhibition of the antioxidant defence link. At the systemic and local levels, patients with EOC experience an imbalance in the cytokine system with a significant activation of the pro-inflammatory component.Conclusion.The disruption of oxidative homeostasis and activation of immune inflammation, detected in patients of late reproductive age with EOC, may lead to the damage of the structural cell components of the functional ovarian tissue, thereby reducing the ovarian reserve.Conflict of interest: the authors declare no conflict of interest.

A laparoscopic approach for removal of ovarian remnant tissue in 32 dogs

BackgroundSurgical treatment of ovarian remnant syndrome (ORS) in dogs usually necessitates large celiotomies and considerable manipulation of organs because of the relatively deep position of ovarian remnant tissue, large patient size, and often encountered adhesions. In women, laparoscopic treatment of ORS is successful and has significant advantages over laparotomy. Since laparoscopic ovariectomy has significant advantages over open ovariectomy in dogs, including reduced surgical stress and postoperative pain and shorter convalescence period, the rationale for a laparoscopic approach of canine ORS is evident. Feasibility and efficacy of a laparoscopic approach for treatment of ORS in dogs was prospectively evaluated using a standardized protocol for diagnosis, treatment, and follow-up. Treatment success was evaluated by histology of removed tissues, postoperative hormone testing, and long-term clinical follow-up.ResultsThirty-two client-owned predominantly medium and large breed dogs diagnosed with ORS underwent abdominal ultrasound for ovarian remnant localization prior to laparoscopic surgery for removal of ovarian remnants. Tissue dissection and excision was performed using a vessel sealing forceps. Laparoscopy subjectively enabled detailed visibility and facilitated detection and removal of suspected ovarian tissue in all cases. Histology confirmed ovarian origin of removed tissue in all dogs. Additionally, a GnRH stimulation test was performed in fourteen dogs after a median follow-up of 10.5 months, which verified absence of residual functional ovarian remnant tissue in all dogs. Median surgery duration was 97.5 min and mean total convalescence duration, subjectively scored by owners, was 1.5 ± 0.7 days. No major complications occurred. Adhesions were observed in 79% of the dogs, complicated the surgical approach, and significantly affected surgery duration (85 versus 109 min; p = 0.03). Minor hemorrhage occurred in 12% and significantly increased surgery duration (95.5 versus 128 min; p = 0.02). Trendelenburg position and lateral tilting of the patient were essential for proper access to ovarian remnants. GnRH stimulation test results and/or absence of clinical signs indicative of ORS after a median follow-up period of 22.5 months confirmed treatment efficacy in all dogs.ConclusionLaparoscopic surgery for ORS in dogs is effective with minimal complications and short convalescence and can successfully replace the conventional, more invasive open surgical procedure.

PROTECTIVE EFFECTS OF CISTANCHES HERBA AQUEOUS EXTRACT ON CISPLATIN-INDUCED PREMATURE OVARIAN FAILURE IN MICE

Background: Chemotherapeutic treatment of premenopausal women has been linked to premature ovarian failure (POF). Cistanches Herba (CH) is a commonly used male impotence and female infertility treatment in China; however, whether CH protects ovaries from chemotherapeutic drug-induced POF remains unclear. In this study, we investigated the protective effects of CH in a mouse model of chemotherapeutic drug-induced POF.&#x0D; Materials and Methods: We administered low- and high-concentration CH to cisplatin-induced POF mice for 2 weeks and determined body and ovarian weights, as well as serum follicle-stimulating hormone (FSH) and estradiol concentrations, to evaluate ovarian function. In addition, we evaluated the protective mechanisms of CH by detecting the levels of apoptosis-related proteins and evaluating markers of mitochondrial function.&#x0D; Results: In POF mice, we observed reduced body and ovarian weights; elevated serum FSH and attenuated estradiol concentrations; apoptosis of ovarian granulosa with concomitant changes in apoptosis-related proteins (including caspase-3, poly adenosine diphosphate-ribose polymerase, Bcl-2, and Bax); and mitochondrial dysfunction, such as a reduction in mitochondrial numbers, destruction of ultrastructural morphology, decrease in ATPase activity, and decreases in mitochondrial membrane potential and mitofusin-2 (a mitochondria dynamin-like GTPase). Significantly, CH reversed, to an extent, functional and morphologic injuries and ovarian tissue apoptosis by up-regulating the level of Mfn2 and the ratio of Bcl-2/Bax. Furthermore, CH reduced cisplatin-induced mitochondrial dysfunction in ovarian tissues.&#x0D; Conclusion: The present findings showed that CH inhibited cisplatin-induced POF through interactions between Mfn2 and Bcl-2/Bax proteins and, possibly, by up-regulation of Mfn2 expression. Ultimately, CH protects ovarian tissues from cisplatin-induced apoptosis.