Selenium mitigates methotrexate-induced testicular injury: Insights from male NMRI mice model.

BACKGROUND AND AIM: Chemotherapy, particularly with methotrexate (MTX), often elicits testicular toxicity, leading to impaired spermatogenesis and hormone imbalances. This study aimed to investigate the potential protective effects of selenium (Se) against MTX-induced testicular injury. MATERIALS AND METHODS: Male mice were divided into control, MTX, Se, and MTX + Se groups. Histopathological examination involved the preparation of testicular tissue sections using the Johnsen's tubular biopsy score (JTBS) for spermatogenesis evaluation. Biochemical tests included the assessment of testosterone, malondialdehyde (MDA), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) levels. Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to analyze the expression of caspase 3 (casp3), tumor protein 53 (p53), B-cell lymphoma 2 (Bcl2), and Bcl2-associated X protein (Bax) genes. Statistical analysis was performed using ANOVA and Tukey's tests (p < .05). RESULTS: Histopathological analysis revealed significant testicular damage in the MTX group, with decreased spermatogenesis and Leydig cell count, while Se administration mitigated these effects, preserving the structural integrity of the reproductive epithelium. Biochemical analysis demonstrated that MTX led to elevated malondialdehyde (MDA) levels and reduced testosterone, LH, and FSH levels, suggesting oxidative stress and Leydig cell dysfunction. Gene expression analysis indicated that MTX upregulated proapoptotic genes (casp3, p53, and bax) while downregulating the antiapoptotic Bcl2 gene. In contrast, Se treatment reversed these trends, highlighting its potential antiapoptotic properties. CONCLUSION: Our findings underscore the potential of Se as a therapeutic agent to mitigate the reproductive toxicity associated with MTX-induced testicular injury. Se exerts protective effects by regulating oxidative stress, preserving hormone balance, and modulating apoptotic pathways. These results suggest that Se supplementation could be a promising strategy to alleviate chemotherapy-induced testicular damage and preserve male fertility.

Melatonin Protects Mouse Type A Spermatogonial Stem Cells against Oxidative Stress via The Mitochondrial Thioredoxin System.

OBJECTIVE: Mitochondrial oxidative stress is an important factor in infertility. The mitochondrial thioredoxin system plays an important role in this condition. N-acetyl-5-methoxy tryptamine (melatonin) plays a role in reducing oxidative stress and apoptosis in spermatogonial stem cells (SSCs). In this study, we explore the probable protective effects of melatonin on the mitochondrial thioredoxin system [thioredoxin 2 (Trx2)/Txnip] in SSCs under oxidative stress. MATERIALS AND METHODS: In this experimental study, SSCs were co-cultured two-dimensionally (2D) with Sertoli cells in DMEM culture medium that contained 10% fetal bovine serum (FBS), 1% antibiotics, and 10 ng/ml glial cell-derived neurotrophic factor (GDNF) for 30 days. The cultured cells were subsequently divided into four groups: control; melatonin (250 µM, 24 hours); melatonin (250 µM, 24 hours)+hydrogen peroxide (H2O2, 50 µM, 24 hours); and H2O2 (50 µM, 24 hours). Intracellular reactive oxygen species (ROS) production was determined by flow cytometry. Malondialdehyde (MDA) levels were measured by Fluorometry. The expressions of apoptotic and antioxidant genes and nuclear factor erythroid 2-related factor 2 (Nrf2), Trx2, and nicotinamide nucleotide transhydrogenase (NNT) proteins were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Adenosine triphosphate (ATP) levels were measured by fluorometry. RESULTS: Melatonin reduced H2O2-induced ROS levels and apoptosis in the SSCs. Melatonin also increased mRNA expression of Nrf2, Trx2, NNT, Sirtuin 3 (Sirt3), and decreased mRNA expression of Txnip, and increased protein expressions of Nrf2, Trx2, NNT thereby increasing activity of the mitochondrial thioredoxin system. In addition, melatonin increased ATP levels. CONCLUSION: Melatonin increased Trx2 expression through the Nrf2 pathway. This study suggests that melatonin may protect SSCs from oxidative stress in diseases related to infertility.

A metabolism targeting three-pronged attack significantly attenuates breast cancer stem cell related markers toward therapeutic application.

Tumor metabolism has provided researchers with a promising window to cancer therapy. The metabolic pathways adopted by cancer cells are different from those of normal cells. Thus, metabolism can be considered a linchpin in targeted cancer therapy. Glycolysis, pentose phosphate pathway, and mitochondria represent three critical metabolic spots with important roles in cancer cell survival and proliferation. In the present study, we aimed to target these pathways using three different inhibitors: 2-deoxyglucose, 6-aminonicotinamide, and doxycycline, separately and in combination. Accordingly, cell viability, lactate production, cell cycle profile, apoptotic profile, and expression of surface and molecular markers of MCF-7 and MDA-MB-231 breast cancer cell lines were investigated under adherent and sphere conditions. Our results from our set conditions indicated various inhibitory effects of these compounds on the breast cancer cell lines. Based on this all-around attack, the combination of drugs demonstrated the most effective inhibitory action compared to separate usage. This study suggests the combined application of these drugs in future investigations and more experimental settings in order to introduce this therapeutic strategy as an efficient anti-cancer treatment.

Brain targeting based nanocarriers loaded with resveratrol in Alzheimer's disease: A review.

Alzheimer's disease (AD) is one of the chief neurological difficulties in the aged population, identified through dementia, memory disturbance, and reduced cognitive abilities. ß-amyloid (Aß) plaques aggregations, generation of reactive oxygen species, and mitochondrial dysfunction are among the major signs of AD. Regarding the urgent need for the development of novel treatments for neurodegenerative diseases, researchers have recently perused the function of natural phytobioactive combinations, such as resveratrol (RES), in vivo and in vitro (animal models of AD). Investigations have shown the neuroprotective action of RES. This compound can be encapsulated by several methods (e.g. polymeric nanoparticles (NPs), solid lipid nanoparticles, Micelles, and liposomes). This antioxidant compound, however, barely crosses the blood-brain barrier (BBB), thereby limiting its bioavailability and stability at the target sites in the brain. Thanks to nanotechnology, the efficiency of AD therapy can be improved by encapsulating the drugs in a NP with a controlled size (1-100 nm). This article addressed the use of RES, as a Phytobioactive compound, to decrease the oxidative stress. Encapsulation of this compound in the form of nanocarriers to treat neurological diseases to improve BBB crossing is also discussed.

Mesenchymal stem cells encapsulation in chitosan and carboxymethyl chitosan hydrogels to enhance osteo-differentiation.

BACKGROUND: Recently biomaterials utilized for designing scaffolds in tissue engineering are not cost-effective and eco-friendly. As a result, we design and develop biocompatible and bioactive hydrogels for osteo-tissue regeneration based on the natural polysaccharide chitosan. Three distinct hydrogel components were used for this. METHODS: Hydrogels networks were created using chitosan 2% (CTS 2%), carboxymethyl chitosan 2% (CMC 2%), and 50:50 mixtures of CTS and CMC (CTS/CMC 50:50). Furthermore, scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy (FTIR), degradation, and swelling behavior of design hydrogels were studied. Also, the cytocompatibility and osteo-differentiation potency were examined by encapsulating mesenchymal stem cells derived from adipose tissue (AMSCs) on the designed hydrogels. RESULTS: According to the findings, our results showed an acceptable pore structure, functional groups, and degradation rate of the designed hydrogels for in vitro evaluation. In addition, employing CMC instead of CTS or adding 50% CMC to the hydrogel component could improve the hydrogel's osteo-bioactivity without the use of external osteogenic differentiation agents. CONCLUSION: The CMC-containing hydrogel not only caused early osteogenesis but also accelerated differentiation to the maturity phase of osteoblasts.

Overview of biological effects of melatonin on testis: A review.

Infertility is a major global health issue and male factors account for half of all infertility cases. One of the causes of male infertility is the loss of spermatogonial stem cells, which may occur because of chemotherapy, radiotherapy or genetic defects. In numerous animal species, the evidence suggests the pineal gland and melatonin secretion in their reproductive activities are involved. Recently, considerable attention has pointed to the usage of melatonin in the treatment of diseases. Melatonin is associated with the regulation of circadian and seasonal rhythmic functions, immune system functions, retinal physiology, spermatogenesis and inhibition of tumour growth in different species. Several studies demonstrated that melatonin acts as an anti-apoptotic, anti-inflammatory, anticancer and antioxidant agent. Melatonin can also protect testicles and spermatogonia against oxidative damage, chemotherapy drugs, environmental radiation, toxic substances, hyperthermia, ischemia/reperfusion, diabetes-induced testicular damage, metal-induced testicular toxicity, improve sperm quality and it affects the testosterone secretion pathway by affecting Leydig cells. Therefore, the objective of this study is to investigate the biological effects of melatonin as a natural antioxidant on testicles and their disorders.

Use of alginate hydrogel to improve long-term 3D culture of spermatogonial stem cells: stemness gene expression and structural features.

The quality and quantity of a spermatogonial stem-cell (SSC) culture can be measured in less time using a 3D culture in a scaffold. The present study investigated stemness gene expression and the morphological and structural characterization of SSCs encapsulated in alginate. SSCs were harvested from BALB/c neonatal mice testes through two-step mechanical and enzymatic digestion. The spermatogonial populations were separated using magnetic-activated cell sorting (MACS) using an anti-Thy1 antibody and c-Kit. The SSCs then were encapsulated in alginate hydrogel. After 2 months of SSC culturing, the alginate microbeads were extracted and stained to evaluate their histological properties. Real-time polymerase chain reaction (PCR) was performed to determine the stemness gene expression. Scanning electron microscopy (SEM) was performed to evaluate the SSC morphology, density and scaffold structure. The results showed that encapsulated SSCs had decreased expression of Oct4, Sox2 and Nanos2 genes, but the expression of Nanog, Bcl6b and Plzf genes was not significantly altered. Histological examination showed that SSCs with pale nuclei and numerous nucleolus formed colonies. SEM evaluation revealed that the alginate scaffold structure preserved the SSC morphology and density for more than 60 days. Cultivation of SSCs on alginate hydrogel can affect Oct4, Sox2 and Nanos2 expression.

Effect of Cerium Oxide Nanoparticles on the Expression of Developmental and Apoptosis Genes of Testicular Tissue in 6-Day-Old NMRI Mice Fetuses.

Cerium oxide (CeO2) has potential applications in medicine and various consumer products. This study investigated the effect of CeO2 on the expression of genes associated with apoptosis and testicular development in mouse embryos. The experimental groups of pregnant mice were injected intraperitoneally with CeO2 at a concentration of 10 mg/kg on days 7 and 14 of pregnancy. Six days after birth, the testicles of neonatal male mice were collected for mRNA expression determination using real-time PCR, protein expression analysis by immunohistochemistry, and apoptotic cell population determination using the TUNEL assay. The results showed that the mRNA expression of the Bax, Caspase-3, and Gsk3-ß genes, unlike the Bcl2 gene, decreased significantly in the experimental group compared to the control group. The expression ratio of Bax/Bcl2 in the experimental group was lower than in the control group. A similar trend was observed in the population of apoptotic cells. In the experimental group, the expression levels of, Gata4, Sox8, and Rad54 at both the mRNA and protein levels increased significantly compared to the control group. Based on the results of this study, CeO2 at a concentration of 10 mg/kg, in addition to producing anti-apoptotic effects on the testicular cells of neonatal mice, can increase the expression of genes involved in testicular development and performance. The current experimental study proved the protective effects of 10 mg/kg CeO2 in developmental and apoptosis genes of testicular tissue in 6-day-old NMRI mice fetuses; however, more experiments are required to evaluate the possible side effects and interactions.

Effects of zinc on expression of apoptosis-related genes in freezing thawing damage of adipose tissue derived mesenchymal stromal cells.

The present study was performed to investigate the effects of zinc supplementation on freezing thawing damage in adipose tissue-derived mesenchymal stromal cells (MSC) of mice through studying cellular viability and gene expression profile of apoptosis. Slow freezing method was conducted and the samples were treated with zinc doses 0, 2.5, 5, 10, 25, 50 and 100 µM. Viability was increased in groups of 2.5, 10 and 25 µM zinc in comparison to the control group. Gene expression study showed that in the group of 2.5 µM zinc, Fas, Bax and Caspase3 had down regulation. Up regulation of Bcl2 was observed in the groups of 10 and 25 µM zinc. P53 did not have a protecting regulation in the groups of study. The present study showed that doses 2.5-25 µM of zinc had a rather safe toxicity, increased cellular viability, and ameliorated expression of apoptosis-related genes in both intrinsic and extrinsic pathways.

The therapeutic effect of cerium oxide nanoparticle on ischaemia/reperfusion injury in rat testis.

Testicular torsion is a dangerous urogenital disorder which is caused by twisting of spermatic cord, and unless immediate treatments happen at a proper time, oxidative stress, occurred during ischaemia reperfusion, finally leads to irreversible disintegration of testicular tissue. One of the first preventive lines is to administrate antioxidant factors. In the present study, we investigate the therapeutic effect of cerium oxide nanoparticle on the injury. We divided 45 rats into nine groups, subjected eight groups to testicular torsion-detorsion, injected different doses of cerium oxide nanoparticle into the peritoneum of six groups and analysed all the groups regarding spermatogenetic indices including sperm count, sperm viability and Johnson mean. Our results showed that cerium oxide nanoparticle can alleviate oxidative stress in testis, and this alleviation promotes the reproductive indices as the concentration of cerium oxide nanoparticles increases. The catalase-mimetic and superoxide dismutase-mimetic activities of cerium oxide nanoparticle are the most probable theories to explain the antioxidant effect of the nanoparticle.

A Stereological Study of the Toxic Effects of Cerium Oxide during Pregnancy on Kidney Tissues in Neonatal NMRI Mice.

BACKGROUND: Both antioxidant and prooxidant activities have been previously reported for cerium oxide (CeO2). The aim of this study was to investigate the effects of CeO2 at different doses on changes in kidney tissues and markers in neonatal mice. METHODS: We randomly divided 30 pregnant NMRI mice into five groups (n = 6 per group)-a control group and four groups treated with intraperitoneal (i.p.) administration of different doses of CeO2 (10, 25, 80, or 250 mg/kg body weight (bw)) on gestation days (GD) 7 and GD14. At the end of the treatment period, we analyzed the kidney tissues and serum samples. The levels of two serum redox markers, malondialdehyde (MDA) and ferric reducing/antioxidant power (FRAP), were determined. Data were analyzed using one-way ANOVA and Tukey's test, and a P value of <0.05 was considered significant. RESULTS: The mean total volumes of the renal corpuscle, glomeruli, and Bowman's capsule membranes significantly increased, and there was a significant decrease in the mean total volume of Bowman's space in the high-dose CeO2 group compared to that in the control group. No statistically significant differences existed in the serum levels of MDA and FRAP in the treated and control groups. CONCLUSION: Our results suggest that high doses of CeO2 impair fetal renal development in pregnant mice, which results in kidney damage. Therefore, CeO2 administration during pregnancy could have dose-dependent adverse effects on the developing kidneys in neonates.

Protective effects of royal jelly on testicular torsion induced ischaemia reperfusion injury in rats.

This study was performed to investigate the protective effects of royal jelly (RJ) on a testicular torsion-induced ischaemia/reperfusion (I/R) injury in adult rats. A total of 40 male Wistar rats were divided into four groups, including 10 rats in each group: Group 1 (sham), Group 2 (Control), group 3 (I/R rats treated with 100 mg/kg RJ for 50 days after torsion) and group 4( I/R rats treated with 20 mg/kg vitamin C for 50 days after torsion). Testicular torsion was created by rotating the right testes 720° a clockwise direction for 90 min. The levels of testosterone were measured by ELISA. Pathological evaluation, mean maturity and quality of the seminiferous tubules were used. Results showed that the testicular histopathology standards and testosterone levels changes were statistically significant in groups 3 and 4. The results obtained in this study may suggest that RJ like vitamin C had protective effects on a testicular ischaemia/reperfusion-induced injury in rats.

Quercetin postconditioning attenuates gastrocnemius muscle ischemia/reperfusion injury in rats.

Quercetin, an antioxidant derived from plants, can play a beneficial role in the protection of various tissues against ischemia-reperfusion injuries (IRI). The purpose of the present research was to investigate the protective effects of quercetin on gastrocnemius muscle ischemia-reperfusion. A total of 80 adult male Wistar rats (weights: 250-300 g) were divided into ten groups (n = 8 per group). We used silk 6.0 surgical thread to create a knit to occlude the femoral artery and vein for 3 hr. The treated groups, which comprised half of each experimental group, received intraperitoneal injections of 150 mg/kg quercetin after the ischemia. Blood flow was subsequently reestablished in the reperfusion phase. The rats were kept in reperfusion for 3, 7, 14, or 28 days after which they were killed with high doses of anesthetic drugs, and the gastrocnemius muscles were removed and fixed. Tissue processing, hematoxylin and eosin and toluidine blue staining, and immunohistochemistry were used to assess tumor necrosis factor-α (TNF-α) and nuclear factor κB (NF-κB) levels. A comparison between treated and untreated ischemic sites showed that on the third day of reperfusion, the severity of edema and NF-κB level decreased significantly; on the 7th day of reperfusion, the severity of edema and the levels of TNF-α and NF-κB decreased significantly; and on the 14th day of reperfusion, all of the parameters showed significant decreases. On the 28th day of reperfusion, there were significantly decreased levels of TNF-α and NF-κB, and decreased mast cell infiltration when compared with the untreated groups. According to the results, administration of quercetin after ischemia could significantly prevent gastrocnemius muscle IRI.

Pre-Implantation Effects of Progesterone Administration on Ovarian Angiogenesis after Ovarian Stimulation: A Histological, Hormonal, and Molecular Analysis.

OBJECTIVE: Progesterone (P4) is known to directly affect ovarian tissue angiogenesis. The present study was designed to show how P4 affects ovarian angiogenesis in hormonal, histological, and molecular levels. METHODS: Fifteen adult female NMRI mice were divided into three groups: Control Group; Case Group I (ovarian stimulation alone); and Case Group II (ovarian stimulation followed by P4 administration). Blood and ovarian tissue samples were assessed for hormonal, histological, and molecular alterations. Gene expression for ovarian vascular endothelium growth factor (VEGF) and hypoxia-inducible factor-1 alpha (HIF-1α) was analyzed using real-time PCR. RESULTS: Ovarian hormone levels were increased in the case groups compared with the control group (p<0.05). Quantitative corpus luteum parameters were increased in the case groups compared with the control group (p<0.05). Quantitative ovarian vascular parameters were significantly different in the case groups compared with the control group. Gene expression analyses shows that the mice in Case Group I had higher levels of ovarian VEGF expression than the mice in the control group (p<0.05). No significant difference in gene expression was observed for HIF-1ɑ. CONCLUSION: Treatment with P4 after ovarian stimulation enhanced ovarian angiogenesis by increasing hormone levels and causing significant structural changes.

The Effect of Cerium Oxide During Pregnancy on the Development of the Testicular Tissue of Newborn NMRI Mice.

Cerium(IV) oxide is widely used as a catalyst in all aspects of human life and human beings are exposed to these materials. The purpose of this experimental study was to investigate the effect of CeO2 during pregnancy on alterations in the testis tissue and blood biochemical parameters in newborn mice. Pregnant NMRI mice were divided randomly into five groups (n = 6 for each group) including one control group and 4 treatment groups. Injection of CeO2 solution was administered intraperitoneally at the doses of 10, 25, 80, and 250 mg/kg.bw, respectively, on GD 7 and GD 14. At the end of treatment period, the testicular histological and biochemical parameters of 2- and 6-day-old newborns were analyzed, as well as the biochemical parameters in serum samples of 15-day-old newborns. The number of spermatogonia, Sertoli, and Leydig cells in the testis of the 2-day-old newborn and spermatogonia and Leydig cells in the testis of the 6-day-old newborns in the 250 mg/kg.bw CeO2 treatment group was significantly reduced compared with the control group (P < 0.05). Testis MDA of the 2- and 6-day-old newborns in the treated group receiving 250 mg/kg.bw of CeO2 was significantly higher than the control group (P < 0.001). There was no significant difference between serum MDA and TAC levels between the treated groups with different doses of CeO2 compared with the control group. Therefore, CeO2 given to dams during pregnancy may affect the testicular tissue and blood biochemical parameters in neonates and may be dose-dependent.

Effect of selenium on freezing-thawing damage of mice spermatogonial stem cell: a model to preserve fertility in childhood cancers.

BACKGROUND: During treatment of childhood cancers, fertility of boys may be affected. Therefore, freezing spermatogonial stem cell (SSC) is recommended. However, freezing-thawing process may cause damage to SSCs. This study was conducted to evaluate protective effects of selenium on freezing-thawing damage of mice SSCs using investigation of cell viability and investigation of apoptosis related genes expression including Fas, Caspase3, Bcl2, Bax and P53. METHODS: SSCs were extracted from 80 6-day-old mice. The SSCs were divided into four groups: cryopreservation along with selenium (low and high dose), vitrification along with selenium (low and high dose), cryopreservation control, and vitrification control. Trypan blue staining and real-time polymerase chain reaction (real-time PCR) were used to investigate cell viability and gene expression, respectively. RESULT: Comparison of cell viability in the experimental groups did not show a significant association. Expression of Fas and Caspase3 was significantly lower in cryopreservation group with low-dose selenium. Expression of Bcl2 was significantly lower in cryopreservation group with high-dose selenium. Expression of Bax and Caspase3 was significantly lower in vitrification group with low-dose selenium, and expression of P53 was significantly upper. Expression of Bax and Fas was significantly lower in vitrification group with high-dose selenium, and expression of P53 was significantly upper (P<0.001). CONCLUSIONS: Selenium had dose dependent effect on apoptosis related genes profile. The only evident effect was the effect of low-dose selenium in cryopreservation on inhibition of apoptosis via extrinsic pathway.

Calendula officinalis stimulate proliferation of mouse embryonic fibroblasts via expression of growth factors TGFß1 and bFGF.

BACKGROUND: TGF-ß has an important role in the process of wound healing and scar formation. The aim of this study is to determine the effects of ethanolic and methanolic extracts of Calendula officinalis on the expression of TGFß1 and bFGF in the mouse embryonic fibroblast cells (MEFs). METHODS: Calendula officinalis extract was purchased and different substances defined with gas chromatography and mass spectrometry. MEFs were prepared and after incubating for 15 min, cell viability analyzed. TGF ß 1 and bFGF gene expression was evaluated by real-time PCR. TGFß1 and bFGF protein expression analyzed by ELISA. The statistical analysis of data was done by using SPSS software. Differences were considered significant at (P < 0.05). RESULTS: The results of the MTT test showed that the concentrations of 5 µg/ml and10 µg/ml were more suitable for cell proliferation. There was an increase in TGF ß 1 gene expression in the MEFs. Expression of TGF ß 1 gene remains the same after 24 h. Gene expression of bFGF showed a similar pattern with TGF ß 1 expression for both solvents. Analysis of TGFß1 protein expression showed an increase in TGFß1 gene expression in the MEFs. Protein expression of bFGF in the MEFs increased at different concentrations at 12 and 24 h after treatment (P < 0.05 and P < 0.01 respectively). CONCLUSION: Calendula officinalis stimulates proliferation of MEFs. Calendula via increased expression of growth factors (TGFß1 and bFGF) at the first 12 h and a decrease of these factors at 24 h after treatment may ameliorate function of the MEFs in the during wound healing.

Effect of royal jelly on testicular antioxidant enzymes activity, MDA level and spermatogenesis in rat experimental Varicocele model.

Varicocele is one of the most prevalent causes of infertility. It causes induction of oxidative stress, increases lipid peroxidation in the testis and disrupts spermatogenesis cycle. The aim of the study was to investigate the possible protective effects of royal jelly against varicocele induced oxidative stress, biochemical and histological alterations in the experimental varicocele model in rat. Twenty-one adult Wistar rats were divided into three groups. The control group (I), Varicocele and administration of normal saline (II), varicocele and treatment with RJ (III). At the end of the experiment, all the animals were sacrificed and testes excised. The activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and MDA levels were measured. Also, histopathological examinations, Johnsen scores and sperm parameters were determined. There was a significant (p<0.05) increase in the activity level of CAT (0.223±0.005), SOD (0.177±0.0062), GPx (9.575±0.318) and a significant (p<0.05) reduction in the MDA level (2.674±0.336) of the experimental varicocele treated with royal jelly when compared to the activity of CAT (0.011±0.004), SOD (0.035±0.0096), GPx (8.864±0.397) and MDA level (4.630±0.579) of the experimental varicocele and administration of normal saline. Results of the Johnsen score showed a significant increase (p<0.05) in the mean score of the RJ group (7.94±1.5) when compared to the normal saline group (6.04±1.4). Therefore, RJ is a potential area for further studies and improving in spermatogenesis cycle after varicocele.

Encapsulation of Satureja khuzistanica extract in alginate hydrogel accelerate wound healing in adult male rats.

BACKGROUND: Finding the best dressing for a specific wound had continued from the past to present. The aim of this study was to evaluate the effect of encapsulated extract of Satureja khuzistanica in hydrogel alginate at wound healing. METHODS: Thirty-two male Wistar rats with a puncture wound in the back of the neck skin were divided randomly into four groups including a control group, Satureja khuzistanica-treated group, hydrogel alginate-treated group, and Satureja khuzistanica encapsulated in hydrogel alginate-treated group. Rats were treated for 22 days. The skin samples were taken on 3rd, 7th, 14th, and 22nd days after treatment for light microscopy. Results were analyzed in accordance with Kruskal-Wallis and Friedman test (for histopathology analysis) by using SPSS v.22 software. RESULTS: Macroscopically evaluations and measurement of wound size showed increased wound healing process in the treated groups. The complete improvement was created on the 14th day. The wound site was not observed on the 22nd day. But the wound site was observed on the 22nd day in the control group. Also, comparison of the percentage of wound healing between the treated and control groups on 3rd, 7th, 14th, and 22nd days showed a significant difference (p < 0.05). Comparison of the H&E stained sections in the studied groups showed that treated groups were effective on wound healing in comparison with the control group. CONCLUSIONS: Encapsulated extract of Satureja khuzistanica in hydrogel alginate may accelerate wound improvement and increase the rate of wound healing without scar formation.

The Effect of Caffeic Acid on Spermatogonial Stem Cell-type A Cryopreservation.

BACKGROUND: Cancer treatment methods can lead to male infertility .in this regard, cryopreservation of spermatogonial stem cells (SSC) and cell-to-person transplantation after the course of treatment to resolve the problem of infertility is a good one. The cryopreservation of SSC is an important process as it can help on the return of spermatogenesis. However, during this process, the stem cells often become damaged which degrades their value for experiments and treatments. Caffeic acid (CA) is an antioxidant that has been shown to increase the viability of cells under stress. The aim of this study was to investigate the effect of CA has on spermatogonial stem cell (SSC) cryopreservation. METHODS: Spermatogonial stem cells isolated from the testes of Balb/c mice pups were cultured in laminincoated dishes, purified using CD90.1 microbeads, then cryopreserved in vitrification media supplemented with 10 µM CA either through a slow or rapid freezing process. After thawing, cell viability was evaluated. Expression of Bax, Fas, Bcl-2 and P53 genes was determined by real-time PCR. Gel electrophoresis was used to confirm the results of the real-time PCR. RESULTS: The viability of the SSCs that were rapidly frozen and treated with CA was observed to be significantly reduced compared to the control group (p < 0.003). The viability SSCs that received CA and underwent the slow freezing treatment was significantly reduced compared to controls (p < 0.002). The expression levels of BAX, BCL-2, and Fas in the rapid freeze-thaw group didn't significantly change. However, the levels of P53 expression were shown to increase. In the group of SSCs that underwent the slow freezing process, the BAX gene expression levels increased, while the levels of BCL-2 gene expression decreased. No significant changes in the level of Fas and P53 expression were detected. When comparing the groups that received CA treatment, SSCs that were rapidly frozen showed an up-regulation of Fas and P53 expression and a down-regulation of Bcl-2 and Bax expression. CONCLUSION: Caffeic acid may protect intact SCCs during the cryopreservation process through stimulating the induction of apoptosis in injured SSCs. Supplementing the vitrification media with CA has a superior effect on the preservation of SSCs.