Evaluating effect of acrylamide and ascorbic acid on oxidative stress and apoptosis in ovarian tissue of wistar rat.

Acrylamide is one of the undesirable compounds created in food, which leads to oxidative stress. Under normal conditions of the body, there is a balance between the production and elimination of free radicals. Imbalance in this process causes oxidative stress. Ascorbic acid has antioxidant properties and can be used to prevent oxidative stress damage. In this study, we considered the effect of acrylamide as a substance that causes oxidative stress and ascorbic acid as an antioxidant. In this experiment, 28 rats were separated into 4 groups (n = 7). Mice were fed orally with acrylamide (10 mg/kg), ascorbic acid (200 mg/kg), both acrylamide and ascorbic acid, and water as AR, AA, AR&AA, and control groups, respectively. Blood and ovarian tissue samples were collected after the final feeding and anesthesia for hematological tests. Blood cells, anti-oxidation status and relative expression of BAX (Bcl-2 Associated X-protein), BCL-2 (B-cell lymphoma-2), Folliculogenesis Specific BHLH Transcription Factor (FIGLA), Follicle Stimulating Hormone Receptor (FSHR), and Klotho (KL) genes were assessed and compared between the study groups. Despite no change in the levels of other factors, white blood cells were significantly different between all groups. The total oxidant and antioxidant index had significant changes in the AR group compared to controls. Glutathione Peroxidase showed the least concentration in the AR group than others although this change was not significant. Gene expression of BAX, BCL-2, FIGLA, FSHR, and KL genes was not significant between the study groups (P > 0.05). The most ratio of BAX to BCL-2 belonged to the AR group compared to other groups. In conclusion, AR probably induces ovarian dysfunction by increasing the proportions of apoptosis-related genes, and the usage of antioxidants like AA can minimize its hazardous effects. However, more research is needed to uncover effective ways to limit AR exposure.

The effects of morphine abuse on sperm parameters, chromatin integrity and apoptosis in men.

OBJECTIVE: Morphine is one of the major psychoactive chemicals in opium that can increase the production of free radicals and thus can negatively affect spermatogenesis. The purpose of this study was to demonstrate the effects of morphine consumption on sperm parameters, DNA integrity and apoptosis in men taking morphine. METHODS: In this case-control study, 30 man abusing morphine (cases) and 30 healthy men (controls) were compared in sperm parameters (count, motility and morphology) and sperm chromatin quality, with Aniline Blue (AB), Toluidine blue (TB) and Chromomycin A3 (CMA3) stains. The participants were matched for age, weight, amount and duration of cigarette smoking. RESULTS: In men with morphine dependency, sperm progressive and total motility (p=0.038 and p=0.000, respectively) showed a significant decrease compared to the control group. Concerning morphine abuse, although morphine can decrease the sperm chromatin condensation and increases the rate of sperm apoptosis, these differences were not statistically significant. CONCLUSIONS: According to our results morphine dependence can reduce male fertility by affecting sperm parameters.

Investigation and Comparison of the Effect of TGF-ß3, kartogenin and Avocado/Soybean Unsaponifiables on the In-vitro and In-vivo Chondrogenesis of Human Adipose-Derived Stem Cells on Fibrin Scaffold.

Due to the lack of suitable therapeutic approaches to cartilage defect, the objective of this study was to determine the effect of Transforming growth factor-ß3 (TGF-ß3), avocado/soybean (ASU) and Kartogenin (KGN) on chondrogenic differentiation in human adipose-derived stem cells (hADSCs) on fibrin scaffold. hADSCs seeded in fibrin scaffold and cultured in chondrogenic media. These cells were divided into 4 groups (control, TGF-ß3, ASU and KGN). Cell viability was estimated by MTT assay. Differentiated cells were evaluated by histological and immunohistochemical (IHC) techniques. Expression genes [sex determining region Y-box 9 (SOX9), Aggrecan (AGG), type II collagen (Coll II) and type X collagen (Coll X)] were assessed by real-time PCR. For a study on an animal model, differentiated cells in fibrin scaffolds were subcutaneously transplanted in rats. Histological and immunohistochemistry were done in the animal model. The results of the real-time PCR indicated that SOX9, AGG and Col II genes expression in TGF-ß3, KGN and ASU groups were significantly higher (p < 0.01) compared to the control group, Col X gene expression only in the TGF-ß3 group was significantly higher (p < 0.01) compared to the control group. The glycosaminoglycan (GAG) deposition was higher in TGF-ß3, KGN and ASU groups compared to the control group. The immunohistological analysis showed the distribution of collagen type X in the extracellular matrix in the fibrin scaffold TGF-ß3 group was significantly higher in control, KGN and ASU groups, and (p < 0.001). ASU, particularly KGN, was suitable for successful chondrogenic differentiation of hADSCs and a suppressor of the consequent hypertrophy.

Protective effects of curcumin on chromatin quality, sperm parameters, and apoptosis following testicular torsion-detorsion in mice.

OBJECTIVE: The chief outcome of testicular torsion in clinical and experimental contexts is testicular ischemia. Curcumin, a compound with anti-inflammatory and antioxidant properties, has fascinated researchers and clinicians for its promise in the treatment of fertility diseases. METHODS: Thirty-five fully grown male mice were randomly classified into five groups: control, sham, testicular torsion, treatment group 1 (testicular torsion+short-term curcumin), and treatment group 2 (testicular torsion+long-term curcumin). Thirty-five days later, spermatozoa from the right cauda epididymis were analyzed with regard to count and motility. Toluidine blue (TB), aniline blue (AB), and chromomycin A3 (CMA3) staining assays were used to evaluate the sperm chromatin integrity. In addition, the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) test was used to assess apoptosis. RESULT: Treatment group 1 exhibited a remarkably elevated sperm count compared to the testicular torsion group. Additionally, notably lower sperm motility was found in the testicular torsion group compared to the control, treatment 1, and treatment 2 groups. Staining (CMA3, AB, and TB) and the TUNEL test indicated significantly greater testicular torsion in the torsion group compared to the control group (p<0.05). The data also revealed notably lower results of all sperm chromatin assays and lower apoptosis in both treatment groups relative to the testicular torsion group (p<0.05). Significantly elevated (p<0.05) AB and TB results were noted in treatment group 1 compared to treatment group 2. CONCLUSION: Curcumin can compensate for the harmful effects of testicular ischemia and improve sperm chromatin quality in mice.

The effects of sesame oil and different doses of estradiol on testicular structure, sperm parameters, and chromatin integrity in old mice.

OBJECTIVE: Studies of the effects of estrogens on the male reproductive system have emphasized the role of these hormones in male fertility. Sesame oil has many phytoestrogenic compounds and may improve male fertility. This study investigated the effects of sesame oil and different concentrations of estrogen on sperm parameters and DNA integrity in male mice. METHODS: Twenty old NMRI (The Naval Medical Research Institute) male mice (40 weeks; weight, 30-35 g) were treated with sesame oil or different concentrations of estrogen (estradiol, 1 and 10 µL/kg/ day) or received no treatment (controls). After 35 days, sperm parameters and DNA integrity were assessed and analyzed. RESULTS: Sperm count, progressive motility, and morphology were decreased in the group that received 10 µL/kg of estradiol. A remarkably lower percentage of DNA fragmentation and protamine deficiency were detected in the group that received 1 µL/kg of estradiol. In the groups that received sesame oil and 1 µL/kg of estradiol, the numbers of spermatogonia and Leydig cells were higher than in controls. The combination of sesame oil and 1 µL/kg of estradiol led to improved sperm parameters and chromatin and testicular structure. CONCLUSION: Based on this study, consumption of sesame oil and a low concentration of estradiol may improve testicular function in older mice.

Comparison of TGF-ß3 and avocado/soybean unsaponifiable on chondrogenesis of human adipose-derived stem cells on poly (lactic-co-glycolic) acid/ hyaluronic acid hybrid scaffold.

OBJECTIVES: Avocado/soybean unsaponifible (ASU) possesses properties including chondroprotective, anticatabolic, and anabolic. The goal behind this research was to detect the effect of ASU and TGF-ß3 on the chondrogenesis of human adipose-derived stem cells (hADSCs) on poly (lactic-co-glycolic) acid (PLGA)/ hyaluronic acid (PLGA/HA) hybrid scaffold. MATERIALS AND METHODS: First hADSCs were seeded in PLGA/Hyaluronic acid scaffold and cultured in chondrogenic media. These cells were assigned into 4 groups: control, TGFß-3, ASU, and TGFß-3+ASU. The viability was assessed separately by MTT. Real-time PCR was used to quantify the expression of chondrogenic specific genes [Sox9, collagen type II (ColII), Aggrecan (AGG)] and collagen type X (ColX). Moreover, Western blotting was employed to evaluate protein expression levels of collagens type II and X. RESULTS: These findings indicated a significant increase in the proliferation and survival of hADSCs differentiated cells by ASU compared with the control group (P=0.008). Real-time PCR results revealed significant differences in the expression of AGG, SOX9, ColII, and ColX genes in the control group when compared with other groups (ASU, TGF-ß3, and TGF-ß3+ASU). ColII protein production significantly dropped in the TGF-ß3 group in comparison with the TGF-ß3+ASU group (0.000). The ColII (P=0.002) and ColX (P=0.002) protein production proved significantly higher in the TGF-ß3+ASU group compared with the ASU group. CONCLUSION: Using the synergist form TGFß-3, ASU induces chondrogenesis in hADSCs in PLGA/HA composite scaffold. This can be deduced with reduction of special markers of hyaline cartilage in comparison with ASU and decreased hypertrophic marker compared with TGF-ß3.

L-Carnitine reduces the negative effects of formalin on sperm parameters, chromatin condensation and apoptosis in mice: An experimental study.

BACKGROUND: Formalin is commonly applied as an antiseptic and tissue fixative. It has reactive molecules that lead to its cytotoxic effects. According to recent studies, formalin causes a change in the testicular and sperm structure and L-carnitine (LC) acts as an antioxidant to counteract its effects. OBJECTIVE: This study aimed to investigate the protective effects of LC on the parameters, chromatin condensation and apoptosis of mice sperm exposed to formalin. MATERIALS AND METHODS: In this experimental study, 24 balb/c mice (25-40 gr,10-12 wk) were divided into three groups (n = 8/each): group I without any injections or gavage; group II, received 10 mg/ kg formalin intraperitoneally (I.P); and group III was exposed to formalin and LC, where a dose of 10 mg/kg formalin was injected I.P daily and LC the dose of 100 mg/kg was kept in a solvent solution. After 31 days, the sperm examination was performed as follows: to evaluate chromatin and DNA quality of the sperm, we applied aniline blue (AB), toluidine blue (TB), chromomycin A3 (CMA3), and terminal transferase-mediated deoxy uridine triphosphate biotin end labeling (TUNEL) tests. RESULTS: Sperm parameters such as count, motility, morphology, and viability displayed a significant decrease in the formalin group. While the data exhibited a considerable augment in sperm parameters in the formalin + LC than the formalin and control groups (p < 0.001), significant differences were detected between groups with respect to TB staining, TUNEL test, CMA3 test and AB staining in the formalin and formalin + LC groups. CONCLUSION: LC can reduce the negative effects of formalin on sperm parameters, chromatin stability, and percentage of apoptosis in an animal model.

Effects of acrylamide in the presence of vitamin E on sperm parameters, chromatin quality, and testosterone levels in mice.

OBJECTIVE: The present study investigated sperm chromatin quality and testosterone levels in acrylamide-treated mice and the possible protective effects of vitamin E on the fertility potential of spermatozoa. METHODS: Thirty-two adult male mice were divided equally into four groups. Group 1 was the control, group 2 received acrylamide (10 mg/kg, water solution), group 3 received vitamin E (100 mg/kg, intraperitoneal), and group 4 received both acrylamide and vitamin E. After 35 days, spermatozoa from the right cauda epididymis were analyzed in terms of count, motility, morphology, and viability. Sperm DNA integrity and chromatin condensation were assessed by acridine orange (AO), aniline blue (AB), toluidine blue (TB), and chromomycin A3 (CMA3) staining. RESULTS: In acrylamide-treated mice, significantly lower sperm concentration, viability, motility, and testosterone levels were found in comparison with the control and acrylamide+vitamin E groups (p<0.05). In the vitamin E group, significantly more favorable sperm parameters and testosterone levels were found than in the other groups (p<0.05). There were also significantly more spermatozoa with less condensed chromatin in the acrylamide-treated mice than in the other groups. Moreover, significantly more spermatozoa with mature nuclei (assessed by AB, CMA3, AO, and TB staining) were present in the vitamin E group than in the control and acrylamide+vitamin E groups. CONCLUSION: This study revealed the deleterious effects of acrylamide on sperm parameters and sperm chromatin quality. Vitamin E can not only compensate for the toxic effects of acrylamide, but also improve sperm chromatin quality in mice.

IMSI procedure improves clinical outcomes and embryo morphokinetics in patients with different aetiologies of male infertility.

Our objective was to evaluate the effect of IMSI on embryo kinetics and clinical outcomes in patients with different aetiologies of male infertility. A total of 150 couples with different aetiologies of male infertility were randomly divided into ICSI and IMSI treatment groups (n = 75). ICSI was done accordingly. For IMSI group, the sperm selection was done using MSOME criteria and then injected. The zygotes were cultured in time-lapse monitoring system (TLM) for 3 days. A total of 650 embryos were developed and assessed using TLM in two groups. Data showed the rate of fragmentation had significant correlation with different aetiologies (p = 0.01), and the timing of s1, t4, s2 and t5 occurred significantly later in oligoasthenoteratozoospermia (OAT) patients compared with others (p < 0.05). In IMSI group, there were no differences in the TLM parameters among different aetiologies (p > 0.05). The rates of chemical pregnancy and implantation (37.8% and 38.2% respectively) were insignificantly higher in OAT patients compare to others (p > 0.05). Also, the clinical pregnancy and live birth rates (32% and 32% respectively) were insignificantly higher in teratozoospermia (T) cases. Sperm selection with MSOME parameters and IMSI can improve the embryo morphokinetics and clinical outcomes in couples with male factor infertility, especially for OAT and T patients.

Vitamin C attenuates detrimental effects of diabetes mellitus on sperm parameters, chromatin quality and rate of apoptosis in mice.

OBJECTIVES: The main goal was to examine the protective effects of vitamin C on sperm parameters, sperm chromatin condensation and apoptosis in experimentally-induced diabetic mice. STUDY DESIGN: 28 adult Syrian mice were divided into 4 groups. In Group1, the mice were diabetic that received a single dose of Streptozocin (STZ) (200mg/kg) intra-peritoneally (ip). Group 2 was included diabetic mice that received vitamin C (10mg/kg/daily, ip). Mice in group 3 were received vitamin C and group 4 was considered as control. After 35 days, sperm analysis was done accordingly. To asses sperm chromatin and DNA quality, we used aniline blue (AB), toluidine blue (TB), chromomycin A3 (CMA3), acridine orange (AO) and terminal transferase mediated deoxyuridine triphosphate biotin end labeling (TUNEL) tests. RESULTS: All of the sperm parameters (count, motility, morphology and viability) had significant reduction in diabetic mice but, the data showed a significant increase in all of the sperm parameters in diabetic+vitamin C when compared with diabetic and control animals (P<0.05). There were significant differences (P<0.001) between groups regarding to TB staining (48.8±5.92 vs 34.3±4.13), AO test (35.9±6.11 vs 20.8±2.89) and TUNEL test (39.42±7.18 vs 22.00±3.65) in diabetic and diabetic+vitamin C groups, respectively. Nevertheless, in CMA3 and AB staining assays, there were not any significant differences between different groups. CONCLUSION: Vitamin C, as a potent antioxidant, can attenuate detrimental effects of diabetes mellitus on the sperm parameters, chromatin quality and apoptosis in an experimental model.

Effects of acrylamide on sperm parameters, chromatin quality, and the level of blood testosterone in mice.

BACKGROUND: Acrylamide (AA) is an important industrial chemical primarily. AA is also found in carbohydrate-rich foods that are prepared at high temperatures, such as French fries and potato chips. It is demonstrated that AA is a carcinogen and reproductive toxin and has ability to induce sperm damage. OBJECTIVE: The aim of this study was to observe the effects of AA on sperm parameters and evaluation of sperm chromatin quality and testosterone hormone in mice. MATERIALS AND METHODS: Totally, 16 adult male mice were divided into two groups. Mice of group A fed on basal diet; group B received basal diet and AA (10 mg/kg, water solution) for 35 days. The right cauda epididymis was incised and then placed in Ham's F10 culture media at 37(o)C for 15 min. Released spermatozoa were used to analyze count, motility, morphology and viability. To determine the sperm DNA integrity and chromatin condensation, the cytochemical techniques including Aniline blue, Acridine orange and Chromomycin A3 staining were used. RESULTS: AA-treated mice had poor parameters in comparison with control animals. In sperm chromatin assessments, except TB (p=0.16), significant differences were found in all of the tests between two groups. It was also seen a significant decrease in concentration of blood testosterone in AA-treated animals when compared to controls (p<0.001). CONCLUSION: According to our results, AA can affect sperm parameters as well as sperm chromatin condensation and DNA integrity in mice. These abnormalities may be related to the reduction in blood testosterone.

Effects of silver nano-particles on sperm parameters, number of Leydig cells and sex hormones in rats.

BACKGROUND: Nano-particles are extensively employed in most industries. Several studies have been started to explore the probable detrimental effects of nano-particles on human reproduction. However, there is insufficient and controversially evident of effects of silver nano-particles on sperm parameters and other reproductive indices. OBJECTIVE: Investigation of the effects of silver nano-particles on sperm parameters, sex hormones and Leydig cells in rat as an experimental model. MATERIALS AND METHODS: In this experimental study, 75 male prepubertal Wistar rats were categorized in five groups including control group and 4 experimental groups (n=15 in each group). The rats in the experimental groups were fed silver nano-particles (60 nm in dimension) with concentrations of 25, 50, 100, and 200 mg/kg/day. After 45 days (about one duration of spermatogenesis in rat), samples of blood were taken from the rats for testosterone, leuteinizing hormone (LH), and follicle stimulating hormone (FSH) assessments. Afterwards, the epididymis and the testis of each rat were dissected for analyzing sperm parameters and Leydig cells. RESULTS: The results demonstrated a statistically significant reduction in number of Leydig cells in experimental groups compared to control one. In addition, the data showed a reduction in testosterone and a rise in LH level which was more obvious in high doses (p<0.05); however, FSH level showed a reduction but it was not statistically significant. A significant decrease was also found in sperm motility and normal sperm morphology in the experimental groups compared to the control one. CONCLUSION: Our results demonstrated that silver nano-particles, in addition to interruption in functions of sex hormones, can diminish the number of Leydig cells and sperm parameter indices. It should be noted that the effects of nano-particles on reproductive indices are dose-dependent.

Effects of experimentally-induced diabetes on sperm parameters and chromatin quality in mice.

BACKGROUND: Diabetes mellitus (DM), primary or idiopathic is a chronic disorder of the carbohydrate, lipid and protein metabolism. DM may impact male reproductive function at several levels. It is shown that DM has detrimental effects on sperm parameters in human and experimental animals. OBJECTIVE: The aim of this study was to observe the effects of diabetes on sperm parameters (viability, count, morphology and motility) and evaluation of sperm chromatin quality in mice. MATERIALS AND METHODS: Totally twenty adult male Syrian mice were divided randomly into 2 groups (n=10). The animals of group A were considered as controls while group B mice were diabetic that received a single dose (200 mg/kg) streptozotocin (STZ) intra peritoneally. After 35 days, the cauda epididymis of each diabetic mouse was dissected and placed in culture medium for 30 min. The swim-out spermatozoa were analyzed for count, motility, morphology and viability. The sperm chromatin quality and DNA integrity, was evaluated with Aniline Blue (AB), Toluidine blue (TB), Acridine orange (AO) and Chromomycin A3 (CMA3) staining. RESULTS: In sperm analysis, the diabetic mice had poor parameters in comparison with control animals (p=0.000). Regarding sperm chromatin quality, the results of TB and AO tests showed statically significant differences between two groups, but in AB and CMA3 staining, we didn't see any differences between them. CONCLUSION: The results showed that STZ-induced diabetes mellitus may influence the male fertility potential via affecting sperm parameters and DNA integrity in mice. However, according to our data, the diabetes doesn't have any detrimental effects on histone-protamines replacement during the testicular phase of sperm chromatin packaging.