Increase In Sperm Concentration Research Articles

Effects of Testicular Platelet-Rich Plasma (PRP) Injection on Sperm Parameters in Men with Severe Oligoasthenoteratozoospermia (OAT): A Clinical Evaluation.

Severe oligoasthenoteratozoospermia (OAT), characterized by a reduced sperm count, motility, and altered morphology, presents a significant challenge in the field of male infertility. Platelet-rich plasma (PRP), renowned for its regenerative capabilities, emerges as a potential intervention for this condition. This study aims to explore the impact of PRP on male infertility, focusing specifically on individuals with severe OAT. The clinical trial study involved 88 infertile men diagnosed with OAT and devoid of underlying diseases. These participants were referred to the infertility center and subsequently divided into two cohorts: a control (44 individuals) and an intervention group (44 individuals). Patients in the intervention group received 2 cc of PRP in each testicle, prepared by centrifuging the patients autologous blood samples. Sperm parameters and DNA fragmentation index (DFI) of the patients were measured before and after the procedure. Statistical analysis used SPSS version 16 software, with a significance level set at less than 5%. The statistical analysis revealed a significant difference in concentration (11.32 ± 8.44 vs. 16.06 ± 15.16, P=0.030), progressive motility (8.86 ± 7.79 vs. 11.97 ± 11.82%, P=0.014) and DNA fragmentation (25.62 ± 12.84 vs. 17.23 ± 9.15%, P<0.001) between the control and intervention groups after PRP injection. However, no significant difference was found in normal morphology (1.63 ± 1.44 vs. 1.81 ± 3.68%, P=0.628) and volume (2.13 ± 0.82 vs. 2.24 ± 1.43, P=0.663) between the control and intervention groups after PRP injection. This study demonstrates the effectiveness of PRP treatment in increasing sperm concentration and motility, while also reducing sperm DNA fragmentation. However, further studies are needed to validate these findings (registration number: IRCT20220317054318N2).

Hydroxytyrosol Ameliorates Busulfan-Induced Oligozoospermia in a Mouse Model

Objective Hydroxytyrosol (HT), a polyphenolic component identified in olive oil and leaves, represents one of the most extensively investigated natural phenol due to its diverse beneficial biological properties. Recent studies have shown that HT exhibits protective effects on reproductive function and can improve sperm quality. However, the precise effect and underlying mechanism of HT on oligozoospermia remain unclear. This study aims to investigate the effect and mechanism of HT across varying concentrations on oligozoospermia. Methods Oligozoospermia was induced in ICR male mice via intraperitoneal injection of busulfan (BUS, 30 mg/kg). The mice were randomly categorized into five groups as Normal, BUS, HT 10 mg/kg, HT 30 mg/kg and HT 50 mg/kg. The efficacy of HT on oligozoospermia was assessed through sperm concentration and motility using a computer-assisted sperm analysis system. Testicular and epididymal histology was examined via hematoxylin-eosin staining, while cytoskeletal integrity was evaluated by immunofluorescence. Hormone levels, along with reactive oxygen species (ROS) and malondialdehyde (MDA) were examined by ELISA kits. Differentially expressed genes were identified through RNA sequencing, and the expression levels of key genes were validated by qPCR. Results The results indicated that HT (50 mg/kg) significantly increased sperm concentration, promoted morphological recovery of testicular tissue, heightened the levels of serum testosterone, follicle-stimulating hormone, luteinizing hormone, as well as decreased the accumulation of ROS and MDA in mice with oligozoospermia. RNA sequencing analysis and RT-qPCR results revealed that HT could restore spermatogenesis and ameliorate oligozoospermia by up-regulating genes related to cilium organization, microtubule-based movement, and spermatid differentiation and development. Conclusions HT demonstrates therapeutic efficacy against busulfan-induced oligozoospermia. The mechanism of HT on oligozoospermia may be related to its influence on cilium and microtubule-based movement, along with the differentiation and development of spermatids. HT shows potential as a prospective therapeutic intervention for the treatment of oligozoospermia.

Brushing rams before and during electroejaculation improves sperm motility and kinetics with slight changes in stress biomarkers

The aim of this study was to determine whether brushing rams before and during electroejaculation (EE) reduces their stress response and improves the characteristics of the ejaculate. A single person brushed each ram for 5min daily, for 15 days, in an individual pen. Semen was collected from five rams brushed before and during EE by the same brusher, while the other five were electroejaculated without being brushed. The treatments were exchanged three days later, so semen was collected from all the rams with both treatments. Brushing increased mass motility (P = 0.05), and curvilinear (P = 0.001), linear (P = 0.02), and average path (P = 0.01) velocities of sperm, as well as the average amplitude of lateral displacement of the sperm head (P = 0.05), and tended to increase sperm concentration (P = 0.09). Brushing tended to reduce the cortisol concentration (P = 0.06) and the duration of head movements when 2V pulse series V was applied (P = 0.1). Brushing increased creatine kinase concentration (P = 0.04) and tended to increase rectal (P = 0.06) and maximum eye surface temperatures (P = 0.1), total time, and number of electrical pulses administered (P = 0.07 for both variables), as well as the sum of pulses per voltage applied during EE (P = 0.06). In rams accustomed to being brushed by the same person, brushing them before and during EE improved semen quality, with slight changes in the stress responses.

Effect of Edible Bird Nest Supplementation Against Busulfan-Induced Oligospermia in Adult Rats

WHO estimates that between 48 million couples and 186 million individuals are facing infertility globally. Male infertility contributes to one-third of the problem with oligospermia as a primary underlying cause. Edible bird nest (EBN), a nutritional natural product, has the potential to improve male fertility status. However, the scientific evidence on oligospermia is sparse. Therefore, this study aimed to evaluate the effectiveness of EBN supplementation in busulfan-induced oligospermia of adult rats. Thirty-six eight-week-old Sprague Dawley male rats (N=36) were randomly and equally divided into six groups (n=6): Control, Sham, Busulfan, EBN 250, 500, and 1000 mg/kg/daily. A single dose of 10 mg/kg busulfan was injected intraperitoneally to induce oligospermia. Daily EBN supplementation was administered for 28 consecutive days, started one day following the busulfan injection. Findings showed that sperm concentration, spermatogenesis index scoring, seminiferous tubule diameter, and germ cell height were reduced in busulfan-induced oligospermia. Meanwhile, the apoptosis status was significantly increased. EBN supplementation had significantly increased sperm concentration, spermatogenesis index scoring, germ cell height, and gene expression of p38 and MAPK14. EBN supplementation also reduced apoptosis in the oligospermia group in a dose-dependent manner. In conclusion, daily EBN supplementation has a protective effect in busulfan-induced oligospermia by increasing sperm production, preserving histological features of the testes, reducing apoptosis, and improving gene expression of p38 and MAPK14.

Can Microfluidics Improve Sperm Quality? A Prospective Functional Study.

The same sperm selection techniques in assisted reproduction clinics have remained largely unchanged despite their weaknesses. Recently, microfluidic devices have emerged as a novel methodology that facilitates the sperm selection process with promising results. A prospective case-control study was conducted in two phases: 100 samples were used to compare the microfluidic device with Density Gradient, and another 100 samples were used to compare the device with the Swim-up. In the initial phase, a significant enhancement in progressive motility, total progressive motile sperm count, vitality, morphology, and sperm DNA fragmentation were obtained for the microfluidic group compared to Density Gradient. Nevertheless, no statistically significant differences were observed in sperm concentration and chromatin structure stability. In the subsequent phase, the microfluidic group exhibited significant increases in sperm concentration, total progressive motile sperm count, and vitality compared to Swim-up. However, non-significant differences were seen for progressive motility, morphology, DNA structure stability, and DNA fragmentation. Similar trends were observed when results were stratified into quartiles. In conclusion, in a comparison of microfluidics with standard techniques, an improvement in sperm quality parameters was observed for the microfluidic group. However, this improvement was not significant for all parameters.

Spermatogenic potential of Chlorophytum borivilianum Santapau & R.R.Fern. in rats

Traditional applications of the roots of Chlorophytum borivilianum Santapau & R.R.Fern. include spermatogenic potential. Using rats as test subjects, the potential of alcoholic extract (AE-SM) and n-Butanol fraction (NB-SM), which is rich in saponin, to alter the levels of testicular testosterone and cholesterol as well as mRNA expression of the 3β-Hydroxy-∆5- steroid dehydrogenase (3β-HSD) and Acute Steroid Regulatory Protein (StAR) were determined. Histological analyses of testicular tissues were performed, and the quantity of sperm in the seminal fluid was measured. The testosterone concentration increased in AE-SM and NB-SM in a dose-dependent and statistically significant manner. In-vivo mRNA expression of the StAR and 3β-HSD enzymes was also enhanced by the test samples. When compared to control animals, AE-SM and NB-SM increased testicular testosterone by 11.46 and 12.98 fold, respectively, while the extract and fractions revealed a 5.72 and 7.70 fold alteration in StAR mRNA expression. In testicular tissues, AE-SM and NB-SM increased 3β-HSD activity by 4.98 and 9.05 fold, respectively, while changing the expression of the 3β-HSD mRNA by 6.46 and 15.54 factor, respectively. The fluid sampled from the caudal region of the AE-SM and NB-SM indicated an increased sperm concentration, and histological examinations revealed dense Sertoli cell cytoplasm and an increase in the number of sperms in the luminal part of the seminiferous tubules. The components of AE-SM and NB-SM might stimulated testosterone biosynthesis in-vivo by modifying mRNA expression of selected steroidogenic enzymes. By modifying the mRNA expression of the StAR and 3β-HSD, AE-SM and NB-SM can raise the level of testosterone in testicular tissues. As evidenced by an elevated sperm concentration discovered within the seminiferous fluid and histological studies, overexpression of steroidogenic enzymes increased testicular testosterone levels and may have stimulated the spermatogenesis process.Saponin enriched n-butanol fraction might have interacted with CREB and additionally stimulated LH receptors to elevate the StAR and 3β-HSD mRNA expression.

Clomiphene Citrate a First Step to Improve Idiopathic Male Infertility: A Retrospective Analysis

Background. Infertility in men is a common and worldwide problem with limited registered treatment options and licensed therapies. Clomiphene citrate (CC) is an empirical off-label therapy. CC is a selective estrogen receptor modulator that may improve testicular function, including sperm quality, by stimulating steroidogenesis and spermatogenesis. However, there is a lack of evidence regarding the efficacy of CC as therapy for male infertility. Objectives. To assess the improvement of CC on sperm parameters, pregnancy, hypogonadism, and measurement of safety. Methods. In this retrospective study, 52 men treated with CC for idiopathic nonobstructive infertility were analyzed. Data of sperm parameters, pregnancy rate, hormonal values, side effects, improvement in hypogonadal symptoms, and potential predictors of treatment response were collected. Results. A total of 52 infertile men underwent CC treatment. An increase in sperm concentration was observed in 19 patients (37%). An upgrade in sperm concentration classification according to the WHO criteria was noted in 21% of the patients receiving CC therapy. Additionally, 15% of initially ineligible patients for intrauterine insemination became eligible during CC. In total, 33% of couples achieved pregnancy, either spontaneously or with assisted reproductive techniques (ART). Low-normal follicle-stimulating hormone and larger testis volume before CC treatment were predictors for better semen concentration during treatment. The retrieval rate was 75% for azoospermic patients who underwent testicular sperm extraction or percutaneous epididymal sperm aspiration, enough for intracytoplasmic sperm injection. With CC, testosterone levels increased and hypogonadal symptoms improved. Mild side effects were reported by 8% of patients. Conclusion. The results of this study contribute to the existing evidence that CC is a safe and noninvasive therapy for idiopathic infertile males. It could be considered as a step-up therapy before initiating more invasive ART procedures.

Effect of Spirulina platensis Supplementation on Reproductive Parameters of Sahrawi and Jabbali Goat Bucks.

Spirulina platensis (SP) is a protein-rich dietary supplement that improves animal reproductive traits. This study investigated the effect of SP supplementation on puberty onset, semen characteristics, scrotal circumference (SC), libido, and hormone concentrations in Sahrawi and Jabbali bucks. The study was conducted in 36 bucks, divided into three groups (n = 6/group), for 70 days. The rations included the following: (1) Control feed (Con) with 14% crude protein and 11.97% MJ/kg DM energy; (2) Con with 2 g SP/head/day SP treatment (T1) and (3) Con with 4 g SP/head/day treatment (T2). The mean (±SEM) SC of both SP groups in the Sahrawi breed was significantly higher (p ≤ 0.05) compared to the Con. The mean of the semen volume significantly increased (p ≤ 0.05) in the SP group than in the Con group in both breeds. SP groups vs. Con groups had increased sperm concentration in Sahrawi bucks than Jabbali bucks. Mean serum luteinizing hormone (LH) and testosterone (Tes) concentrations in Jabbali bucks were significantly higher (p ≤ 0.05) in the SP groups compared to Sahrawi bucks. SP improved the SC, semen quality, libido, sperm concentration, and LH and Tes concentrations in both breeds. The results of the current study suggest that adding SP to the diet may have the ability to improve the semen quality of the local Omani bucks.

Clomiphene Citrate in the Management of Infertility in Oligospermic Obese Men with Hypogonadism: Retrospective Pilot Study.

Background and Objectives: Obesity is a significant risk factor for hypogonadism and infertility that is further associated with reduced semen quality. The aim of this study is to evaluate the effect of clomiphene citrate (CC), prescribed for treating infertility, on serum testosterone and semen parameters, particularly in oligospermic obese hypogonadal men. Materials and Methods: A retrospective analysis of data related to men (n = 53) who underwent CC treatment for infertility and hypogonadism (testosterone < 300 ng/dL) was performed. Patients with obesity (BMI ≥ 30 kg/m2) and sperm concentration ≤ 15 × 106/mL were included for analysis. Results: The overall results showed that, in oligospermic obese men (n = 31), treatment with CC significantly improved baseline sperm concentration (4.5 ± 6.8 × 106/mL vs. 11.4 ± 15.5 × 106/mL, p < 0.05) and motility (31.5% ± 21.5% vs. 42.6% ± 14.7%, p < 0.05). Furthermore, subsequent examination of oligospermic hypogonadal obese men treated with CC (n = 13) revealed substantial improvements in baseline serum testosterone levels (193.8 ± 59.3 ng/dL vs. 332.7 ± 114.8 ng/dL, p < 0.05) along with an increase in sperm concentration, total motility, and normal morphology. Conclusions: The results of this retrospective study suggest that CC treatment not only improves chances of fertility outcomes by substantially improving semen parameters but also increases total serum testosterone levels in oligospermic obese men without any supplemental and expensive testosterone replacement therapy.

OR25-02 Hunting The Magic Bullet In Treating Male Secondary Hypogonadism Associated With Metabolic Syndrome: Results Of a Prospective Study Comparing Liraglutide Vs. Gonadotropins

Abstract Disclosure: S. La Vignera: None. R.A. Condorelli: None. A.E. Calogero: None. R. Cannarella: None. E. Greco: None. A. Aversa: None. Background: Previously, we hypothesized a role for glucagon-like peptide-1 (GLP1) analogs in the treatment of male infertility (DOI: 10.1111/andr.13015), although no studies have so far investigated this issue. Purpose: To prospectively investigate the effects of treatment with liraglutide, a GLP1 analog, on reproductive and sexual function in men with functional hypogonadism of childbearing age. Patients and Methods: 110 obese men of childbearing age (18-35 years) with functional hypogonadism were enrolled and divided into three groups based on their desire to have children. Group A consisted of men actively seeking paternity, Group B of men who did not seek paternity, and Group C of men who had already fathered a child. Group A was treated with gonadotropins [urofollitropin 150 IU three times a week and human chorionic gonadotropin (hCG) 2000 IU twice a week], Group B with liraglutide (3 mg daily), and Group C with transdermal testosterone (60 mg daily). The following endpoints were measured at enrolment and after 4 months: anthropometric and metabolic parameters, gonadotropins, total testosterone, conventional sperm parameters (Group A and B only), the score of the International Index of Erectile Function 5 Items (IIEF5), and frequency of phosphodiesterase type 5 inhibitors (PDE5i) use. Results: The groups did not differ on any of the baseline endpoints. Weight, body mass index, and waist circumference significantly decreased after treatment in Group B, but not in the other Groups. Liraglutide significantly increased LH (3.2±0.2 IU/L vs. 2.0±0.3 IU/L; p&amp;lt;0.01), FSH (2.6±0.2 IU/L vs. 1.6±0.3 IU/L; p&amp;lt;0.01), total testosterone (4.1±0.5 ng/mL vs. 1.4±0.6 ng/mL; p&amp;lt;0.05), and sex hormone-binding globulin (36.0±4.0 nmol/L vs. 14.0±3.0 nmol/L) levels from baseline, while no improvement was observed in the other Groups. Liraglutide increased sperm concentration (16±4 mil/ml vs. 6±2 mil/ml; p&amp;lt;0.05), progressive motility (34±4 % vs. 14±2 %; p&amp;lt;0.01), and morphology (10±2 % vs. 4±2 %; p&amp;lt;0.05), while treatment with gonadotropins only increased sperm progressive motility (19±2 % vs. 12±3 %; p&amp;lt;0.05) compared to baseline. The IIEF-5 score improved after liraglutide treatment (4±2 vs. 21±4; p&amp;lt;0.05), while no significant changes were observed in the other Groups. Similarly, PDE5i use decreased significantly in Group B (31%), compared with Groups A (48%) and C (45%). Conclusions: Liraglutide is an effective weight loss drug, which also improves serum gonadotropin and total testosterone levels, sperm conventional parameters, and sexual function in obese patients with functional hypogonadism seeking parenthood. This is the first study documenting the efficacy of liraglutide on the testicular function of these patients. Presentation Date: Saturday, June 17, 2023

Predictive role of 17α-hydroxy-progesterone serum levels of response to follicle-stimulating hormone in patients with abnormal sperm parameters

To study the possible role of serum 17α-hydroxy-progesterone (17αOH-P) levels in predicting favorable responses to follicle-stimulating hormone (FSH) administration in patients with normal serum FSH levels and idiopathic abnormal sperm parameters. Prospective cohort study. University-affiliated fertility center. Fifty patients with oligozoospermia, asthenozoospermia, and/or teratozoospermia and normal serum levels of gonadotropins and total testosterone (TT). Treatment with exogenous FSH is administered subcutaneously at a dose of 150 IU 3 times a week for 3 consecutive months. Luteinizing hormone levels, FSH levels, TT levels, 17αOH-P levels, testicular volume, conventional sperm parameters, and seminal spermatid concentration were evaluated before and after therapy. To evaluate the predictive role of pretreatment serum 17αOH-P levels on FSH responsiveness, the doubling of sperm concentration at the end of the FSH administration was considered a positive outcome. After therapy, patients showed a significant increase in sperm concentration, total sperm count (TSC), progressive motility, percentage of normal forms, FSH levels, TT levels, and testicular volume. There was a negative correlation between pretreatment 17αOH-P levels and the posttreatment increase in sperm concentration, TSC, progressive motility, and normal morphology, and a positive correlation with the posttreatment increase in spermatids. Predictive analysis showed that 17αOH-P levels (<1.18 ng/mL) foretold a doubling of sperm concentration with a sensitivity of 90.0% and a specificity of 73.3%, and of TSC with a sensitivity of 91.3% and a specificity of 81.48%. The results of this study suggest that pretreatment serum levels of 17αOH-P, a marker of steroidogenic function, appear to be able to predict the success of subcutaneous administration of exogenous FSH in terms of spermatogenesis improvement. Receiver operating characteristic curves indicated that 17αOH-P levels (<1.18 ng/mL) predict a doubling of sperm concentration and TSC after exogenous FSH administration to patients with idiopathic abnormal sperm parameters and normal gonadotropin levels.

Micronutrient supplements as antioxidants in improving sperm quality and reducing DNA fragmentation

BackgroundSpermatogenesis and sperm quality may be negatively impacted by an increase in reactive oxygen species. This study investigates the efficacy of combined antioxidant therapy for treating male infertility, as measured by semen analyses and the sperm DNA fragmentation index (DFI). Infertile men with a high sperm DNA fragmentation index were instructed to take two oral micronutrient capsules daily for three months. Each antioxidant formulation contained 60 mg vitamin E, 400 µg folic acid, 30 mg selenium, 125 mg L-arginine, 220 mg L-carnitine, 7.5 mg coenzyme Q10, 40 mg L-glutathione, and 20 mg zinc citrate. At entry and post-treatment, the general characteristics, semen analysis, and sperm chromatin dispersion assays were recorded and compared.ResultsAfter three months of treatment with antioxidant compounds, the quality of spermatozoa improved significantly, as indicated by a decrease in the mean DNA fragmentation index from 45.6 ± 17.2% to 34.8 ± 20.3%; an increase in sperm concentration from 29.7 × 106/mL to 35.7 × 106/mL (p < 0.001), an increase in a total number of spermatozoa from 72.1 × 106 to 95.5 × 106 (p = 0.012), and an increase in the vitality from 75.5 ± 17.1 to 81.1 ± 14.4% viable forms (p < 0.001).ConclusionsMicronutrient supplementation can improve sperm quality and DNA integrity in infertile men. Men with infertility and significant sperm DNA fragmentation who take antioxidants for three months experience a reduction in DNA fragmentation index and an increase in sperm quality as measured by the semen analysis.Trial registrationNCT04509583. Registered 12 August 2020, Hue University of Medicine and Pharmacy Ethics Committee—Retrospectively registered.

Strontium Chloride Improves Reproductive Function and Alters Gut Microbiota in Male Rats.

Strontium (Sr) is an essential trace element in the human body and plays an important role in regulating male reproductive health. Recent studies have shown that gut flora plays a key role in maintaining spermatogenesis, as well as testicular health, through the gut-testis axis. At present, it is unclear whether gut microbiota can mediate the effects of Sr on sperm quality, and what the underlying mechanisms may be. We investigated the effects of different concentrations of strontium chloride (SrCl2) solutions (0, 50, 100, and 200 mg/kg BW) on reproductive function and gut microbiota in male Wistar rats (6-8 weeks, 250 ± 20 g). All the animals were euthanized after 37 days of treatment. The Sr-50 group significantly increased sperm concentration, sperm motility, and sperm viability in rats. After Sr treatment, serum and testicular testosterone (T) and Sr levels increased in a dose-dependent manner with increasing Sr concentration. At the same time, we also found that testicular marker enzymes (ACP, LDH) and testosterone marker genes (StAR, 3β-HSD, and Cyp11a1) increased significantly in varying degrees after Sr treatment, while serum NO levels decreased significantly in a dose-dependent manner. Further investigation of intestinal flora showed that SrCl2 affected the composition of gut microbiome, but did not affect the richness and diversity of gut microbiota. Sr treatment reduced the number of bacteria with negative effects on reproductive health, such as Bacteroidetes, Tenericutes, Romboutsia, Ruminococcaceae_UCG_014, Weissella, and Eubacterium_coprostanoligenes_group, and added bacteria with negative effects on reproductive health, such as Jeotgalicoccus. To further explore the Sr and the relationship between the gut microbiota, we conducted a Spearman correlation analysis, and the results showed that the gut microbiota was closely correlated with Sr content in serum and testicular tissue, sex hormone levels, and testicular marker enzymes. Additionally, gut microbiota can also regulate each other and jointly maintain the homeostasis of the body's internal environment. However, we found no significant correlation between intestinal flora and sperm quality in this study, which may be related to the small sample size of our 16S rDNA sequencing. In conclusion, the Sr-50 group significantly increased T levels and sperm quality, and improved the levels of testicular marker enzymes and testosterone marker genes in the rats. Sr treatment altered the gut flora of the rats. However, further analysis of the effects of gut microbiota in mediating the effects of SrCl2 on male reproductive function is needed. This study may improve the current understanding of the interaction between Sr, reproductive health, and gut microbiota, providing evidence for the development of Sr-rich foods and the prevention of male fertility decline.

Improvement of spermatozoa concentration due to maximal exercise with Vitis gracilis Wall.

Context: Swimming is a common form of exercise; however, excessive exercise might reduce sperm count by lowering testosterone levels and increasing the production of free radicals, commonly known as reactive oxygen species (ROS). In Indonesia, Vitis gracilis Wall. is a traditional remedy for increasing stamina. Aims: To assess the concentration of spermatozoa after vigorous physical activity and V. gracilis administration, as well as the histological and apoptotic changes in testicular histology that occur via caspase-3 expression. Methods: This study was conducted on six groups of rats: the control group (G+), a group of rats subjected to vigorous swimming then administered 0.2 mg/kg BW vit C (GVitC), and three groups of rats subjected to vigorous swimming then administered 100, 125, or 150 mg/kg BW V. gracilis (G100, G125, and G150). Testicular tissue and blood serum samples were extracted from the rats subjected to vigorous swimming. Testicular tissue was immunohistochemically stained using caspase-3 antibody and TUNEL assays, while blood samples were analysed using ELISA. Results: V. gracilis administration significantly affected IL-6 and testosterone levels (p&lt;0.00). Testosterone had a greater impact on spermatozoa concentration than IL-6. Caspase-3 expression and the proportion of apoptotic cells were both markedly reduced. Conclusions: Administering 125 mg/kg BW V. gracilis can help to increase sperm concentration by reducing apoptosis through altering caspase-3 and IL-6 levels, thereby preventing inflammation. This plant might be a viable molecular therapeutic target for staminal medicines.

Extended one-generation reproductive toxicity study evaluating gardenia blue in Sprague Dawley rats

Gardenia blue powder was administered at 0.5%, 2.5%, or 5.0% in feed to male and female Sprague Dawley rats in an Extended One-Generation Reproductive Toxicity Study (OECD Test Guideline 443). The dosed diet began 14 days before mating and was continued at the same concentration level for the entire study for all parental animals (P0) and offspring (F1). At weaning, offspring were allocated into one of 5 cohorts for different endpoints. P0 and F1 animals had blue urine, blue or black feces, and blue discolorations in gastrointestinal organs, mesenteric lymph nodes, and kidneys. This treatment-related finding was not considered adverse as there were no histopathologic correlates. There was a dose-related increase in sperm concentration in P0 and F1 males. There were dose-related increases in heart weights of F1 postnatal day (PND) 21 males, male and female thyroid weights, and female TSH levels of PND 91 F1 offspring, with no histopathological correlate. There were no consistent treatment-related adverse effects on any other parameters evaluated for general toxicity, reproductive toxicity, developmental neurotoxicity, or developmental immunotoxicity. The highest dietary concentration (5.0%) of gardenia blue powder was the no observed adverse effect level (NOAEL) for male and female rats at all life stages evaluated.

Wuzi Yanzong administration alleviates Sertoli cell injury by recovering AKT/mTOR-mediated autophagy and the mTORC1–mTROC2 balance in aging-induced testicular dysfunction

Ethnopharmacological relevanceWuzi Yanzong Prescription (WZ), a classic traditional Chinese medicine formula, has the properties of kidney nourishing and essence strengthening, and it is widely used to treat male infertility with a long history. Sertoli cells are injured with aging, resulting in testicular dysfunction, and WZ effectively rejuvenates the age-related decline of testicular function. However, whether the therapeutic effects of WZ on aging-related testicular dysfunction are dependent on the restoration of Sertoli cell function remains unclear. Aim of the studyIn a mouse model of natural aging, we explored the protective effects of WZ and its potential mechanisms. Materials and methodsFifteen-month-old C57BL/6 mice were randomized to receive either standard diet or WZ (2 and 8 g/kg) for 3 months. Meanwhile, 10 1-month-old mice were considered the adult control group and received standard diet for 3 months. The testis and epididymis were rapidly collected, and the sperm quality, testicular histology, Sertoli cell numbers, tight junction (TJ) ultrastructure, and blood–testis barrier-associated protein expression and localization were assessed. ResultsWZ significantly increased sperm concentration and sperm viability, improved the degenerative histomorphology and elevated the seminiferous epithelium height. Furthermore, WZ increased the number of Sertoli cells, restored the ultrastructure of the Sertoli cell TJ, and upregulated the expression of TJ-associated proteins (zonula occludens-1 and Claudin11), ectoplasm specialized-associated proteins (N-Cadherin, E-Cadherin and β-Catenin), and gap junction-associated protein (connexin 43), but did not affect the expression of Occludin and cytoskeletal protein (Vimentin). In addition, WZ did not change the localization of zonula occludens-1 and β-Catenin in aged testis. Moreover, WZ increased the expression of autophagy-associated proteins (light chain 3 beta and autophagy related 5) and decreased the expression of p62, phosphorylated mammalian target of rapamycin, and phosphorylated AKT in Sertoli cells. Finally, we found that WZ attenuated mTOR complex 1 (mTORC1) activity and upregulated mTORC2 activity, as evidenced by inhibition of the expression of the regulatory-associated protein of mTOR, phosphorylated p70 S6K, and phosphorylated ribosomal protein s6 and enhancement of the expression of Rictor in the Sertoli cells of aging mice. ConclusionsWZ improves the injury of Sertoli cells by restoring AKT/mTOR-mediated autophagy and the mTORC1–mTROC2 balance in Sertoli cells during aging. Our findings provide a new mechanism of WZ in the treatment of aging-induced testicular dysfunction.

Effect of oestrogen modulation on semen parameters in men with secondary hypogonadism: Systematic review and meta-analysis.

Selective oestrogen receptor modulators and aromatase inhibitors stimulate endogenous gonadotrophins and testosterone in men with hypogonadism. There are no systematic reviews/meta-analyses assessing the effects of selective oestrogen receptor modulators/aromatase inhibitors on semen parameters in men with secondary hypogonadism. To assess the effect of monotherapy or a combination of selective oestrogen receptor modulators/aromatase inhibitors on sperm parameters and/or fertility in men with secondary hypogonadism. A systematic search was conducted in PubMed, MEDLINE, Cochrane Library and ClinicalTrials.gov. Study selection and data extraction were performed by two reviewers independently. Randomised controlled trials and non-randomised studies of interventions reporting effects of selective oestrogen receptor modulators and/or aromatase inhibitors on semen parameters or fertility in men with low testosterone with low/normal gonadotrophins were selected. The risk of bias was assessed using ROB-2 and ROBINS-I tools. The results of randomised controlled trials were summarised using vote counting while summarising effect estimates where available. Non-randomised studies of intervention meta-analysis were conducted using the random-effect model. The certainty of evidence was assessed using GRADE. Five non-randomised studies of interventions (n=105) of selective oestrogen receptor modulators showed an increase in sperm concentration (pooled mean difference 6.64 million/mL; 95% confidence interval 1.54, 11.74, I2 =0%) and three non-randomised studies of interventions (n=83) of selective oestrogen receptor modulators showed an increase in total motile sperm count (pooled mean difference 10.52; 95% confidence interval 1.46-19.59, I2 =0%), with very low certainty of evidence. The mean body mass index of participants was >30kg/m2 . Four randomised controlled trials (n=591) comparing selective oestrogen receptor modulators to placebo showed a heterogeneous effect on sperm concentration. Three included men with overweight or obesity. The results were of very low certainty of evidence. Limited pregnancy or live birth data were available. No studies comparing aromatase inhibitors with placebo or testosterone were found. Current studies are of limited size and quality but suggest that selective oestrogen receptor modulators may improve semen parameters in those patients, particularly when associated with obesity.

P-102 Inhibition of RANKL increases the number of motile sperm in a subgroup of infertile men

Abstract Study question Does inhibition of RANKL increase semen quality in infertile men? Summary answer Overall, inhibition of RANKL (Denosumab) did not affect semen quality. However, in a subgroup of men with high serum AMH sperm production was significantly increased. What is known already Currently, there is no treatment available for male infertility, but studies in animal and preclinical human data have suggested a role for RANKL in male reproduction. Study design, size, duration A single-center, double-blinded, randomized clinical trial in 100 infertile men randomly assigned (1:1) to s.c. injection with 60 mg/mL Denosumab or s.c. saline injection 1 mL (0.9%) (placebo). The primary endpoint was changes in semen production, though the study was designed as an explorative study, with the aim of investigating the profile of which infertile men who would have a beneficial treatment outcome of Denosumab. Participants/materials, setting, methods All participants were infertile men referred to our andrological clinic due to impaired semen quality. Blood and semen samples were collected at baseline and after one and two cycles of spermatogenesis on day 80 and day 160, respectively. Main results and the role of chance There were no differences in semen quality or reproductive hormones at day 80 or day 160 between men treated with Denosumab or placebo. However, men with a serum AMH ≥ 40 pmol/L had a significant increase in total motile sperm (143% (SD 116%); p = 0.011), progressive motile sperm (133% (SD 100%); p = 0.039) and a borderline increase in sperm concentration (64% (SD 95%); p = 0.056) at day 80 compared to baseline. Further stratifying men according to both serum AMH (≥ 40 pmol/L) and serum RANKL (highest 50%), showed a significantly higher increase in sperm concentration (107% (SD 111%) vs. -8% (SD 68%); p = 0.009), total motile sperm (256% (SD 270%) vs. 31% (SD 125%); p = 0.039) and progressive motile sperm (191% (SD 194%) vs. 18% (SD 134%) p = 0.013) at day 80 in men treated with Denosumab compared to placebo treated men. Furthermore, serum FSH (-28% (SD 33%) vs. 0%, (SD 22%); p = 0.041) and serum LH (-29% (SD 28%) vs. 20% (SD 51%), p = 0.048) were significantly lower and testosterone/LH ratio (50% (SD 51%) vs. -7% (SD 43%); p = 0.024) significantly higher at day 80 in men treated with Denosumab compared to placebo treatment. Limitations, reasons for caution This is a report from a randomized clinical trial, with an explorative design. Randomized clinical trials in a predefined group of infertile men with high serum AMH, are needed to explore if inhibition of RANKL improves semen quality and pregnancy rate in infertile couples with male factor infertility. Wider implications of the findings The increase in sperm production after Denosumab treatment in infertile men with preserved Sertoli cell function needs to be validated, in a trial selecting men based on serum AMH. Ultimately, Denosumab may be a potential breakthrough in treating male infertility at least in some infertile men with high serum AMH. Trial registration number NCT03030196

P-026 Effect of Microsurgical Varicocele Ligation in Patients with Isolated Oligozoospermia

Abstract Study question Is varicocelectomy effective in improving the semen quality of patients with isolated oligozoospermia (IO)? Summary answer Varicocelectomy is a valid option for infertile men with clinical varicocele and documented IO considering the significant improvement in semen concentration. What is known already Varicocele is the leading correctable cause of male factor infertility and its surgical treatment has been associated with sound improvement in semen quality and fertility potential. While varicocelectomy is clearly indicated in men with abnormal semen parameters, its effect on men with isolated abnormalities is less commonly assessed. Study design, size, duration The charts of 1521 patients who underwent microsurgical subinguinal varicocelectomy for clinically palpable varicocele between 2011-2021 were retrospectively reviewed. Infertile patients with IO were included in the study group, while data from patients with oligoasthenoteratozoospermia (OAT) were utilized for comparison. Participants/materials, setting, methods Men with azoospermia, a history of genitourinary infection, and those utilizing peri-operative fertility medications were excluded. The collected data included patient's demographics, varicocele grade and veins’ sizes, and their semen (semen analysis, WHO 2010; sperm DNA fragmentation, Halosperm) and hormone parameters before and up to 6 months after varicocelectomy. Data were analyzed using Wilcoxon Signed Ranks test and chi-squared tests. A p value &amp;lt;0.05 was considered significant. Main results and the role of chance A total of 96 patients with IO and 123 patients with OAT were included in the study. Patients with IO had a mean age of 32.5 ± 7 years, and an average testis volume of 15.6ml. The varicocele was of grades 2 and 3 in 58% and 38% of patients. Among the IO group, a significant increase in sperm concentration was identified postoperatively (p = 0.01). Surprisingly, results revealed a significant decrease in sperm total motility, however, the postoperative median was still within the normal range (45.5%). No significant changes were observed in other semen or hormone parameters. When a comparison was made with the OAT group, 83% of patients with IO had an improvement in sperm concentration compared to 63.9% of patients in OAT group following varicocelectomy (P = 0.044). However, a higher percentage of patients showed an improvement in total motility among the OAT group (67.6%) in comparison to the IO group (42.9%) (p = 0.04). Limitations, reasons for caution Results were obtained from a relatively small sample size. However, varicocele treatment is not necessarily considered as the first treatment option in men with mild or isolated derangements in semen quality. Wider implications of the findings Results of this study shed the light on the effect of varicocele treatment in men with isolated oligozoospermia and hence may be beneficial for patient counselling in the clinic. Trial registration number not applicable

P-010 The impact of COVID-19 Vaccination on Sperm Parameters in Indian Men: A Comparative Study

Abstract Study question To analyse the effect of COVID-19 vaccines on sperm parameters in Indian men seeking infertility treatment. Summary answer COVID-19 vaccination did not impact the sperm parameters of Indian men undergoing infertility treatment in this study. What is known already The COVID-19 pandemic triggered an expedited response from the scientific community to develop preventive vaccine programs against the deadly SARS-CoV-2 virus. Although COVID-19 is predominantly a respiratory system illness, initial reports suggested that it could have a short-term negative impact on the sperm parameters of the males affected by the disease too. Misinformation led to unfounded concerns about the potential consequences of the vaccines on male fertility, causing a great deal of initial vaccine hesitancy. To address these concerns, we compared the sperm parameters in men before and after COVID vaccination available in India at the time of this study. Study design, size, duration It was a multi-centre, prospective, observational study of 960 male participants conducted at our fertility clinics between June 2021 to December 2022. The study participants were matched for age and had no known risk factors. We excluded 267 participants from this study since they did not meet the inclusion criteria, bringing our total study population down to 693. Participants/materials, setting, methods The study participants provided two semen samples for analysis before and after receiving any of the COVID-19 vaccines available in India at the time of this study. A trained embryologist analysed the pre-vaccination and post-vaccination sperm concentration and motility of these males, following the 6th Edition of WHO Manual for Semen Analysis. GraphPad Prism 9 was used for statistical analysis of the data &amp; p ≤ 0.05 was considered statistically significant. Main results and the role of chance The study enrolled 693 people, with 96.5% receiving at least one dose and 90.35% receiving two or more. The most common vaccines used among the group were Indian made-Covishield and Covaxin, received by 71.8% &amp; 24.3% of the participants, respectively. In the group of participants who had tested positive for COVID (n = 259), the sperm concentration increased by 3.29 million/ml and the total motility increased by 3.24% post-vaccination. However, these changes were not statistically significant (p = 0.140 and p = 0.099, respectively). In the group of participants who had not tested positive for COVID (n = 424), we observed a similar increase in sperm concentration and total motility post-vaccination. The sperm concentration increased by 1.52 million/ml and the total motility increased by 0.85%. However, these changes were also not statistically significant (p = 0.580 and p = 0.404, respectively). Subgroup analysis of different vaccines did not reveal any statistically significant differences either. In conclusion, the study found that COVID-19 vaccination, including the type of vaccine used, did not significantly impact sperm parameters in men. Limitations, reasons for caution Supply chain disruptions in the COVID vaccination program meant we could not match the duration between the pre-vaccination and the post-vaccination semen analyses in this study. Additionally, the findings are specific to our study population and may not be generalizable to other populations or settings. Wider implications of the findings This study found that COVID-19 vaccination did not affect the chances of Indian men at fatherhood. Our work also supports the safety of Indian-made vaccines for men seeking infertility treatment, as it is one of the most extensive studies in this area. Trial registration number Not Applicable