Background: Diminished ovarian reserve (DOR) is a clinical syndrome with reproductive and endocrine disorders. This study aimed to examine the effect of crocin on oxidative stress, gene expression, oocyte maturation, and embryo quality in DOR patients who underwent a controlled ovarian hyperstimulation (COH) cycle.Methods: As a clinical trial, this study involved 34 DOR patients trying to conceive by assisted reproductive technique who were divided into two groups (17 each): An intervention group receiving crocin (15 mg, once daily, for 12 weeks) and a control group receiving a placebo (tablets with the same form of the drug). Pre- and post-intervention, demographic information was gathered, and hormonal levels (follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2)) were measured. In the subsequent COH cycle, oocyte maturation, embryo quality, level of both superoxide dismutase (SOD) and reactive oxygen species (ROS) in follicular fluid, expression of GDF9, BMP15, and Nrf2 genes in granulosa cells were measured.Results: The collected data as a comparison between groups showed alteration of criteria in the intervention group as follows: Significant reduction of FSH (P<0.01), increased level of SOD in the follicular fluid (P<0.0001), decreased level of oxidative stress in the granulosa cells (P<0.0001), increased expression of Nrf2 gene (P<0.08), and of GDF9, BMP15 genes (P<0.0001) in the granulosa cells. The rate of oocyte maturation and embryo quality were significantly higher in the intervention group compared to the control group (P<0.05 and P<0.005, respectively).Conclusion: Our study discussed how the Krocina supplement can slow down the progression of the disease by reducing the level of FSH, and oxidative stress, increasing the maturation rate of oocytes, and increasing the quality of embryos in women with DOR. Further research is needed to investigate the effect of crocin in improving fecundity for women with DOR.
Abstract In the present study hydroxyapatite (HA) and HA/nanosilica (NS) slurries were separately prepared by adding precipitated HA to distilled water and to colloidal silica suspension, respectively and their rheological properties were compared to each other. Both slurries where then dried, powdered, compacted as disks and sintered at 1 100–1 300°C to evaluate and compare their physical, mechanical and some biological properties. The results showed that the HA/NS slurry was more stable and thixotropic than the pure HA slurry. X-ray diffraction analysis showed that the sintered HA/NS was a multi-phase material composed of apatite, tricalcium phosphate, cristobalite and amorphous glass, depending on sintering temperature. Both mechanical and biological properties of the composite were considerably better than those of sintered HA. Bioactivity of the composite was confirmed by precipitation of apatite nanocrystals onto the surfaces of the sample after soaking in simulated body fluid. The results of cell culture tests showed the same proliferation rate of rat calvaria osteoblasts on both sintered HA and composite with higher alkaline phosphatase activity for the latter. The results indicated that the composite with improved mechanical and biological properties may act more successfully than HA as a bone scaffold material.
In this study, we present an electrospun gelatin (EG) scaffold to mimic the extracellular matrix of the testis. The EG scaffold was synthesized by electrospinning and crosslinked with glutaraldehyde vapor to decrease its water solubility and degradation rate. The scanning electron microscope micrographs showed the homogenous morphology of randomly aligned gelatin fibers. The average diameter of gelatin fibers before and after crosslinking was approximately 180 and 220 nm, respectively. Modulus, tensile strength, and elongation at break values were as 161.8 ± 24.4 MPa, 4.21 ± 0.54 MPa, and 7.06 ± 2.12 MPa, respectively. The crosslinked EG showed 75.2% ± 4.5% weight loss after 14 days with no changes in the pH value of degradation solution. Cytobiocompatibility of the EG for sertoli cells and embryonic stem cells (ESCs) was determined in vitro. Sertoli cells were isolated from mouse testis and characterized by immunostaining and flow cytometry. The effects of EG on proliferation and attachment of both sertoli cells and ESCs were examined. The EG scaffolds exhibited no cytotoxicity for sertoli and ESCs. Both sertoli and ESCs were well attached and grown on EG. Coculture of sertoli and ESCs on EG showed better ESCs adhesion compared with ESCs alone. Our findings indicate the potential of EG as a substrate for proliferation, adhesion, and coculture of sertoli and ESCs and may be considered as a promising engineered microenvironment for in vitro coculture system with the aim of guiding stem cells differentiation toward sperm-producing cells.
This study aimed to investigate the protective effect of Gallic acid (GA) on the Cyclophosphamide (CP) toxicity induced in the reproductive system.After a pilot study for dose responses of Gallic acid ,Forty adult male NMRI mice were divided into 5 groups (n=8): control, sham (NaCl Serum: 0.2mL per day), CP (15 mg kg-1 per week; IP), GA (12.5 mg kg-1 per day ; IP) and GA (12.5 mg kg-1 per day ; IP) +CP(15 mg kg-1 per week; IP). After treatment, the left testis was detached and used for Histological examination and right testis used for Malondialdehyde (MDA) measures. Left caudal epididymis was placed in the Ham's F10 medium and released spermatozoa were used in order to analyze sperm parameters. Sperm DNA fragmentation was assessed by Sperm Chromatin Dispersion (SCD) method.In the CP group, there was a significant increase in the sperm DNA fragmentation (% 57.89 ± 23.91) compared with control group (% 24.52 ± 10.27). That was significantly improved by GA (12.5 mg kg-1 per day ; IP) in GA+CP group (% 28.4 ± 8.85) compared to CP group (p< .001).A significant increase was reported about MDA levels in CP group (6.26 ± 2.59) in compared with the control group (4.30 ± 2.05), But GA (3.24 ± 1.33) decreased it in GA+ CP group (p< .01). The histopathological investigation revealed marked testicular atrophy in CP group, whereas GA diminished these deviations (P< .05).Gallic acid can modify the reproductive toxicity of cyclophosphamide in NMRI mice and increase the antioxidant capacity of testis tissue.
The current study was conducted to assess the relationship between testicular cells in spermatogenesis, through which the production of healthy and mature sperm is essential. However, it seems necessary to obtain more information about the three-dimensional pattern of the testis cells arrangement, which is directly related to the function of the testis after induction of diabetes. Twelve adult mice (28-30 g) were assigned into two experimental groups: (1) control and (2) diabetic (40 mg/kg STZ). The epididymal sperm collected from the tail of the epididymis and testes samples were taken for stereology, immunocytochemistry and RNA extraction. Our data showed that diabetes could notably decrease the number of testicular cells, together with a reduction of total sperm count. In addition, the results from the second-order stereology indicated the significant changes in the spatial arrangement of Sertoli cells and spermatogonial cells in the diabetic groups, in comparison with the control (P < .05). Moreover, the immunohistochemistry results showed a significant reduction in Sex-determining Region Y (SRY) box 9 gene (SOX9), vimentin, occludin, and connexin-43 positive cells in the diabetic groups compared with the control (P < .05). Furthermore, our data showed that the expression of steroidogenic acute regulatory protein steroidogenic acute regulatory protein (StAR) and peripheral benzodiazepine receptor peripheral benzodiazepine receptor (PBR) was significantly reduced in the diabetic groups, in comparison with the control (P < .05). These findings suggest that structural and functional changes of testis cells after induction of diabetes cause the alterations in the spatial arrangement of Sertoli and spermatogonial cells, ultimately influencing the normal spermatogenesis in mice.
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